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1
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Iqbal AJ, Krautter F, Blacksell IA, Wright RD, Austin-Williams SN, Voisin MB, Hussain MT, Law HL, Niki T, Hirashima M, Bombardieri M, Pitzalis C, Tiwari A, Nash GB, Norling LV, Cooper D. Galectin-9 mediates neutrophil capture and adhesion in a CD44 and β2 integrin-dependent manner. FASEB J 2021; 36:e22065. [PMID: 34847625 DOI: 10.1096/fj.202100832r] [Citation(s) in RCA: 24] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/21/2021] [Revised: 10/29/2021] [Accepted: 11/10/2021] [Indexed: 01/20/2023]
Abstract
Neutrophil trafficking is a key component of the inflammatory response. Here, we have investigated the role of the immunomodulatory lectin Galectin-9 (Gal-9) on neutrophil recruitment. Our data indicate that Gal-9 is upregulated in the inflamed vasculature of RA synovial biopsies and report the release of Gal-9 into the extracellular environment following endothelial cell activation. siRNA knockdown of endothelial Gal-9 resulted in reduced neutrophil adhesion and neutrophil recruitment was significantly reduced in Gal-9 knockout mice in a model of zymosan-induced peritonitis. We also provide evidence for Gal-9 binding sites on human neutrophils; Gal-9 binding induced neutrophil activation (increased expression of β2 integrins and reduced expression of CD62L). Intra-vital microscopy confirmed a pro-recruitment role for Gal-9, with increased numbers of transmigrated neutrophils following Gal-9 administration. We studied the role of both soluble and immobilized Gal-9 on human neutrophil recruitment. Soluble Gal-9 significantly strengthened the interaction between neutrophils and the endothelium and inhibited neutrophil crawling on ICAM-1. When immobilized, Gal-9 functioned as an adhesion molecule and captured neutrophils from the flow. Neutrophils adherent to Gal-9 exhibited a spread/activated phenotype that was inhibited by CD18 and CD44 neutralizing antibodies, suggesting a role for these molecules in the pro-adhesive effects of Gal-9. Our data indicate that Gal-9 is expressed and released by the activated endothelium and functions both in soluble form and when immobilized as a neutrophil adhesion molecule. This study paves the way for further investigation of the role of Gal-9 in leukocyte recruitment in different inflammatory settings.
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Affiliation(s)
- Asif J Iqbal
- Institute of Cardiovascular Sciences (ICVS), College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK
| | - Franziska Krautter
- Institute of Cardiovascular Sciences (ICVS), College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK
| | - Isobel A Blacksell
- The William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK
| | - Rachael D Wright
- The William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK
| | - Shani N Austin-Williams
- The William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK
| | - Mathieu-Benoit Voisin
- The William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK
| | - Mohammed T Hussain
- Institute of Cardiovascular Sciences (ICVS), College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK
| | - Hannah L Law
- The William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK
| | - Toshiro Niki
- Research Division, GalPharma Company, Ltd., Kagawa, Japan
| | - Mitsuomi Hirashima
- Department of Immunology and Immunopathology, Faculty of Medicine, Kagawa University, Kagawa, Japan
| | - Michele Bombardieri
- The William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK
| | - Costantino Pitzalis
- The William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK
| | - Alok Tiwari
- Department of Vascular Surgery, University Hospitals Birmingham, Birmingham, UK
| | - Gerard B Nash
- Institute of Cardiovascular Sciences (ICVS), College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK
| | - Lucy V Norling
- The William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK
| | - Dianne Cooper
- The William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK
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2
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Fujimura T, Yamada Y, Umeyama T, Kudo Y, Kanamori H, Mori T, Shimizu T, Kato M, Kawaida M, Hosoe N, Hasegawa Y, Matsubara K, Shimojima N, Shinoda M, Obara H, Naganuma M, Kitagawa Y, Hoshino K, Kuroda T. Maintenance treatment with infliximab for ulcerative ileitis after intestinal transplantation: A case report. World J Clin Cases 2021; 9:5270-5279. [PMID: 34307578 PMCID: PMC8283613 DOI: 10.12998/wjcc.v9.i19.5270] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/03/2021] [Revised: 03/18/2021] [Accepted: 05/17/2021] [Indexed: 02/06/2023] Open
Abstract
BACKGROUND Evidence has been published on the successful applications of the anti-tumor necrosis factor alpha antibody infliximab, such as induction therapy, salvage treatment for acute cellular rejection, and treatment for chronic ulcerative inflammation, in intestinal transplant recipients. However, the optimal protocol for the effective use of infliximab remains largely undetermined due to scarcity of available clinical data. We report a continuative application of infliximab as maintenance therapy for recurrent chronic ulcerative ileitis in a recipient of isolated intestinal transplantation (ITx). CASE SUMMARY The patient was a 11-year-old boy with intestinal motility disorder classified as a hypogenic type of intestinal dysganglionosis. The patient underwent living-donor related intestinal transplant. His immunosuppression regimen consisted of daclizumab, tacrolimus, and steroids. Although he did not show rejection while on tacrolimus monotherapy, routine screening endoscopy showed several ulcerative lesions in the distal end of the graft 2 years after the intestinal transplant. Endoscopic work up to evaluate the progression of anemia revealed stenosis with ulcerative inflammatory changes and multiple longitudinal ulcers in the graft. Since the endoscopic findings suggested ulcerative lesions in Crohn's disease, infliximab treatment was considered. Treatment with infliximab and a small dose of oral prednisolone afforded successful withdrawal of total parenteral nutrition and maintenance of a well-functioning graft without infectious complications for 5 years since the administration of the first dose of infliximab. CONCLUSION Infliximab is effective as maintenance therapy for recurrent chronic ulcerative ileitis in an isolated ITx patient.
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Affiliation(s)
- Takumi Fujimura
- Department of Pediatric Surgery, National Saitama Hospital, Wako Shi, Saitama 351-0102, Japan
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Yohei Yamada
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Tomoshige Umeyama
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Yumi Kudo
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Hiroki Kanamori
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Teizaburo Mori
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Takahiro Shimizu
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Mototoshi Kato
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Miho Kawaida
- Department of Pathology, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Naoki Hosoe
- Center for Diagnostic and Therapeutic Endoscopy, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Yasushi Hasegawa
- Department of Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Kentaro Matsubara
- Department of Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Naoki Shimojima
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Masahiro Shinoda
- Department of Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
- Digestive Diseases Center, International University of Health and Welfare, Mita Hospital, Tokyo 108-8329, Japan
| | - Hideaki Obara
- Department of Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Makoto Naganuma
- Department of Gastroenterology and Hepatology, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Yuko Kitagawa
- Department of Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Ken Hoshino
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
| | - Tatsuo Kuroda
- Department of Pediatric Surgery, Keio University School of Medicine, Tokyo 160-8582, Japan
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3
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Kempers L, Sprenkeler EGG, van Steen ACI, van Buul JD, Kuijpers TW. Defective Neutrophil Transendothelial Migration and Lateral Motility in ARPC1B Deficiency Under Flow Conditions. Front Immunol 2021; 12:678030. [PMID: 34135903 PMCID: PMC8202084 DOI: 10.3389/fimmu.2021.678030] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/08/2021] [Accepted: 05/13/2021] [Indexed: 12/11/2022] Open
Abstract
The actin-related protein (ARP) 2/3 complex, essential for organizing and nucleating branched actin filaments, is required for several cellular immune processes, including cell migration and granule exocytosis. Recently, genetic defects in ARPC1B, a subunit of this complex, were reported. Mutations in ARPC1B result in defective ARP2/3-dependent actin filament branching, leading to a combined immunodeficiency with severe inflammation. In vitro, neutrophils of these patients showed defects in actin polymerization and chemotaxis, whereas adhesion was not altered under static conditions. Here we show that under physiological flow conditions human ARPC1B-deficient neutrophils were able to transmigrate through TNF-α-pre-activated endothelial cells with a decreased efficiency and, once transmigrated, showed definite impairment in subendothelial crawling. Furthermore, severe locomotion and migration defects were observed in a 3D collagen matrix and a perfusable vessel-on-a-chip model. These data illustrate that neutrophils employ ARP2/3-independent steps of adhesion strengthening for transmigration but rely on ARP2/3-dependent modes of migration in a more complex multidimensional environment.
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Affiliation(s)
- Lanette Kempers
- Molecular Cell Biology Laboratory, Department of Molecular and Cellular Haemostasis, Sanquin Research, Amsterdam University Medical Center (AUMC), Amsterdam, Netherlands
| | - Evelien G G Sprenkeler
- Department of Blood Cell Research, Sanquin Research, AUMC, University of Amsterdam, Amsterdam, Netherlands.,Department of Pediatric Immunology, Rheumatology and Infectious Diseases, Emma Children's Hospital, AUMC, University of Amsterdam, Amsterdam, Netherlands
| | - Abraham C I van Steen
- Molecular Cell Biology Laboratory, Department of Molecular and Cellular Haemostasis, Sanquin Research, Amsterdam University Medical Center (AUMC), Amsterdam, Netherlands
| | - Jaap D van Buul
- Molecular Cell Biology Laboratory, Department of Molecular and Cellular Haemostasis, Sanquin Research, Amsterdam University Medical Center (AUMC), Amsterdam, Netherlands.,Leeuwenhoek Centre for Advanced Microscopy, Section Molecular Cytology, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, Netherlands
| | - Taco W Kuijpers
- Department of Blood Cell Research, Sanquin Research, AUMC, University of Amsterdam, Amsterdam, Netherlands.,Department of Pediatric Immunology, Rheumatology and Infectious Diseases, Emma Children's Hospital, AUMC, University of Amsterdam, Amsterdam, Netherlands
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4
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Pérez-Figueroa E, Álvarez-Carrasco P, Ortega E, Maldonado-Bernal C. Neutrophils: Many Ways to Die. Front Immunol 2021; 12:631821. [PMID: 33746968 PMCID: PMC7969520 DOI: 10.3389/fimmu.2021.631821] [Citation(s) in RCA: 119] [Impact Index Per Article: 29.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/21/2020] [Accepted: 02/08/2021] [Indexed: 12/21/2022] Open
Abstract
Neutrophils or polymorphonuclear leukocytes (PMN) are key participants in the innate immune response for their ability to execute different effector functions. These cells express a vast array of membrane receptors that allow them to recognize and eliminate infectious agents effectively and respond appropriately to microenvironmental stimuli that regulate neutrophil functions, such as activation, migration, generation of reactive oxygen species, formation of neutrophil extracellular traps, and mediator secretion, among others. Currently, it has been realized that activated neutrophils can accomplish their effector functions and simultaneously activate mechanisms of cell death in response to different intracellular or extracellular factors. Although several studies have revealed similarities between the mechanisms of cell death of neutrophils and other cell types, neutrophils have distinctive properties, such as a high production of reactive oxygen species (ROS) and nitrogen species (RNS), that are important for their effector function in infections and pathologies such as cancer, autoimmune diseases, and immunodeficiencies, influencing their cell death mechanisms. The present work offers a synthesis of the conditions and molecules implicated in the regulation and activation of the processes of neutrophil death: apoptosis, autophagy, pyroptosis, necroptosis, NETosis, and necrosis. This information allows to understand the duality encountered by PMNs upon activation. The effector functions are carried out to eliminate invading pathogens, but in several instances, these functions involve activation of signaling cascades that culminate in the death of the neutrophil. This process guarantees the correct elimination of pathogenic agents, damaged or senescent cells, and the timely resolution of the inflammation that is essential for the maintenance of homeostasis in the organism. In addition, they alert the organism when the immunological system is being deregulated, promoting the activation of other cells of the immune system, such as B and T lymphocytes, which produce cytokines that potentiate the microbicide functions.
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Affiliation(s)
- Erandi Pérez-Figueroa
- Unidad de Investigación en Inmunología y Proteómica, Hospital Infantil de México Federico Gómez, Secretaría de Salud, Mexico City, Mexico
- Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad Universitaria, Mexico City, Mexico
| | - Pablo Álvarez-Carrasco
- Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad Universitaria, Mexico City, Mexico
| | - Enrique Ortega
- Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad Universitaria, Mexico City, Mexico
| | - Carmen Maldonado-Bernal
- Unidad de Investigación en Inmunología y Proteómica, Hospital Infantil de México Federico Gómez, Secretaría de Salud, Mexico City, Mexico
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5
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Jin Y, Jones L, Gorbet M. Investigation of the response of tear-film neutrophils to interleukin 8 and their sensitivity to centrifugation, fixation, and incubation. Sci Rep 2020; 10:19690. [PMID: 33184318 PMCID: PMC7665065 DOI: 10.1038/s41598-020-75806-y] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/28/2019] [Accepted: 10/19/2020] [Indexed: 11/26/2022] Open
Abstract
During eye closure, a large number of neutrophils (polymorphonuclear neutrophils, PMNs) invade the ocular surface and are often referred to as tear-film PMNs. While immunophenotyping experiments have been performed on tear-film PMNs, the impact of commonly used experimental procedures on their phenotype as well as their response to interleukin-8 (IL-8), a physiological inflammatory mediator, have not yet been investigated. A gentle eye wash method was used to collect cells at home. In the morning upon awaking, participants washed their eyes with sterile phosphate buffer saline (PBS) and collected the runoff into a sterile polypropylene tube. The cell collection was then delivered to the lab within two hours. The effects of centrifugation, incubation and fixation with paraformaldehyde (PFA) before (pre-fixed staining) or after (post-fixed staining) incubation with antibodies were characterized. Tear-film PMNs as well as blood PMNs (used for comparison) were also stimulated with IL-8. To assess the reproducibility of cell collection and variability in receptor expression over time, participants were also asked to collect cells three times over a period of a month. The change in expression of surface receptors, CD11b, CD16, CD55, CD66b, important inflammatory and activation markers, and CD45 (PAN leukocyte marker) was assessed by flow cytometry. Fixing tear-film PMNs prior to the staining with antibodies resulted in a significant (fivefold or more) reduction in the expression of CD11b, CD16 and CD45 when compared to unfixed samples, while CD16 was the only receptor to undergo significant downregulation upon post-staining fixation. Furthermore, additional centrifugation step prior to antibody incubation as well as long (4 h) incubation at 37 °C resulted in significant reductions in expression of CD11b, CD16 and CD55 when compared to control samples. As opposed to blood PMNs, stimulating tear-film PMNs with IL-8 did not induce any significant changes in expression of CD11b, CD16, CD55 and CD66b. When working with collected tear-film PMNs, our results suggest that any additional centrifugation and incubation step should be avoided, or at least limited, and post fixation staining is recommended in order to preserve cell phenotype and cell integrity of tear film PMNs. Our study also adds further information on the reproducibility of the gentle eye wash as well as the inability of tear-film PMNs to modulate their surface receptors upon stimulation with IL-8. The latter may be due to prior exposure to IL-8, activation in the closed-eye environment, or a reduced ability to respond to inflammatory stimulus. Further mechanistic studies will be needed to gain a better understanding of the tear-film neutrophil phenotype.
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Affiliation(s)
- Yutong Jin
- School of Optometry and Vision Science, University of Waterloo, Waterloo, Canada
- Centre for Ocular Research and Education, University of Waterloo, Waterloo, Canada
| | - Lyndon Jones
- School of Optometry and Vision Science, University of Waterloo, Waterloo, Canada
- Centre for Ocular Research and Education, University of Waterloo, Waterloo, Canada
| | - Maud Gorbet
- School of Optometry and Vision Science, University of Waterloo, Waterloo, Canada.
- Department of Systems Design Engineering, University of Waterloo, Waterloo, Canada.
- Centre for Ocular Research and Education, University of Waterloo, Waterloo, Canada.
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6
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Shi JH, Luo L, Chen XL, Pan YP, Zhang Z, Fang H, Chen Y, Chen WD, Cao Q. Real-world cost-effectiveness associated with infliximab maintenance therapy for moderate to severe Crohn’s disease in China. World J Gastroenterol 2020; 26:6455-6474. [PMID: 33244205 PMCID: PMC7656205 DOI: 10.3748/wjg.v26.i41.6455] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/02/2020] [Revised: 08/05/2020] [Accepted: 09/17/2020] [Indexed: 02/06/2023] Open
Abstract
BACKGROUND Infliximab was the first approved biologic treatment for moderate to severe Crohn’s disease (MS-CD) in China. However, the cost-effectiveness of infliximab maintenance therapy (IMT) for MS-CD relative to conventional maintenance therapy remained unclarified.
AIM To assess the cost-effectiveness of IMT for MS-CD in Chinese patients from the perspective of Chinese public insurance payer.
METHODS A cohort of MS-CD patients managed in a Chinese tertiary care hospital was created to compare IMT with conventional maintenance therapy (CMT) for clinical outcomes and direct medical costs over a 1-year observation time using conventional regression analyses. A decision-analytic model with the generated evidence was constructed to assess the cost-effectiveness of IMT relative to CMT using reimbursed medical costs.
RESULTS Based on the included 389 patients, IMT was associated with significantly higher disease remission chance [odds ratio: 4.060, P = 0.003], lower risk of developing new complications (odds ratio: 0.527, P = 0.010), higher utility value for quality of life (coefficient 0.822, P = 0.008), and lower total hospital costs related to disease management (coefficient -0.378, P = 0.008) than CMT. Base-case cost-effectiveness analysis estimated that IMT could cost Chinese health insurance payers ¥55260 to gain one quality-adjusted life year (QALY). The cost-effectiveness of IMT was mainly driven by the estimate of quality of life, treatment efficacy of maintenance therapy, mortality risk associated with active disease, and unit price of infliximab. The probability that IMT was cost-effective at a willingness-to-pay threshold of three times gross domestic product [2018 Chinese gross domestic product per capita (GDPPC)] was 86.4%.
CONCLUSION IMT significantly improved real-world health outcomes and cost the Chinese public health insurance payers less than one GDPPC to gain one QALY in Chinese MS-CD patients.
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Affiliation(s)
- Ji-Hao Shi
- Department of Gastroenterology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou 310016, Zhejiang Province, China
| | - Liang Luo
- Department of Gastroenterology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou 310016, Zhejiang Province, China
| | - Xiao-Li Chen
- Department of Gastroenterology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou 310016, Zhejiang Province, China
| | - Yi-Peng Pan
- Department of Gastroenterology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou 310016, Zhejiang Province, China
| | - Zhou Zhang
- Department of Gastroenterology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou 310016, Zhejiang Province, China
| | - Hao Fang
- Department of Gastroenterology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou 310016, Zhejiang Province, China
| | - Ying Chen
- Department of Project, Changsha Normin Health Technology Ltd, Changsha 410013, Hunan Province, China
| | - Wen-Dong Chen
- Department of HEOR, Normin Health Consulting Ltd, Toronto L5R 0E9, Ontario, Canada
| | - Qian Cao
- Department of Gastroenterology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou 310016, Zhejiang Province, China
- Inflammatory Bowel Disease Center, Sir Run Run Shaw Hospital, Hangzhou 310016, Zhejiang Province, China
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7
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Activated neutrophils exert myeloid-derived suppressor cell activity damaging T cells beyond repair. Blood Adv 2020; 3:3562-3574. [PMID: 31738831 DOI: 10.1182/bloodadvances.2019031609] [Citation(s) in RCA: 79] [Impact Index Per Article: 15.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/14/2019] [Accepted: 09/02/2019] [Indexed: 12/20/2022] Open
Abstract
Myeloid-derived suppressor cells (MDSCs) have the capacity to suppress T-cell-mediated immune responses and impact the clinical outcome of cancer, infections, and transplantation settings. Although MDSCs were initially described as bone marrow-derived immature myeloid cells (either monocytic or granulocytic MDSCs), mature neutrophils have been shown to exert MDSC activity toward T cells in ways that remain unclear. In this study, we demonstrated that human neutrophils from both healthy donors and cancer patients do not exert MDSC activity unless they are activated. By using neutrophils with genetically well-defined defects, we found that reactive oxygen species (ROS) and granule-derived constituents are required for MDSC activity after direct CD11b-dependent interactions between neutrophils and T cells. In addition to these cellular interactions, neutrophils are engaged in the uptake of pieces of T-cell membrane, a process called trogocytosis. Together, these interactions led to changes in T-cell morphology, mitochondrial dysfunction, and adenosine triphosphate depletion, as indicated by electron microscopy, mass spectrometry, and metabolic parameters. Our studies characterize the different steps by which activated mature neutrophils induce functional T-cell nonresponsiveness and irreparable cell damage.
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8
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Anti-TNF Therapy in Crohn's Disease. Int J Mol Sci 2018; 19:ijms19082244. [PMID: 30065229 PMCID: PMC6121417 DOI: 10.3390/ijms19082244] [Citation(s) in RCA: 181] [Impact Index Per Article: 25.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/26/2018] [Revised: 07/04/2018] [Accepted: 07/07/2018] [Indexed: 02/06/2023] Open
Abstract
Crohn’s disease (CD) accounts for a variety of clinical manifestations or phenotypes that stem from chronic inflammation in the gastrointestinal tract. Its worldwide incidence is increasing including younger or childhood-onset of disease. The natural history of Crohn’s disease is characterized by a remitting and relapsing course that progresses to complications and surgery in most patients. The goals of treatment are to achieve clinical and endoscopic remission, to avoid disease progression and minimise surgical resections. Medical treatment usually features antibiotics, corticosteroids, immunomodulators (thiopurines, methotrexate). Anti-TNF (tumour necrosis factor) therapy was approved for use in Crohn’s disease in 1998, and has changed the paradigm of treatment, leading to improved rates of response and remission in patients. There are significant considerations that need to be borne in mind, when treating patients including immunogenicity, safety profile and duration of treatment.
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9
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Roy J, Mazzaferri J, Filep JG, Costantino S. A Haptotaxis Assay for Neutrophils using Optical Patterning and a High-content Approach. Sci Rep 2017; 7:2869. [PMID: 28588217 PMCID: PMC5460230 DOI: 10.1038/s41598-017-02993-6] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/05/2016] [Accepted: 04/21/2017] [Indexed: 12/30/2022] Open
Abstract
Neutrophil recruitment guided by chemotactic cues is a central event in host defense against infection and tissue injury. While the mechanisms underlying neutrophil chemotaxis have been extensively studied, these are just recently being addressed by using high-content approaches or surface-bound chemotactic gradients (haptotaxis) in vitro. Here, we report a haptotaxis assay, based on the classic under-agarose assay, which combines an optical patterning technique to generate surface-bound formyl peptide gradients as well as an automated imaging and analysis of a large number of migration trajectories. We show that human neutrophils migrate on covalently-bound formyl-peptide gradients, which influence the speed and frequency of neutrophil penetration under the agarose. Analysis revealed that neutrophils migrating on surface-bound patterns accumulate in the region of the highest peptide concentration, thereby mimicking in vivo events. We propose the use of a chemotactic precision index, gyration tensors and neutrophil penetration rate for characterizing haptotaxis. This high-content assay provides a simple approach that can be applied for studying molecular mechanisms underlying haptotaxis on user-defined gradient shape.
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Affiliation(s)
- Joannie Roy
- Research Center, Maisonneuve-Rosemont Hospital, Montreal, Quebec, Canada.,Biomedical Engineering Institute, University of Montreal, Montreal, Quebec, Canada
| | - Javier Mazzaferri
- Research Center, Maisonneuve-Rosemont Hospital, Montreal, Quebec, Canada
| | - János G Filep
- Research Center, Maisonneuve-Rosemont Hospital, Montreal, Quebec, Canada.,Department of Pathology and Cell Biology, University of Montreal, Montreal, Quebec, Canada
| | - Santiago Costantino
- Research Center, Maisonneuve-Rosemont Hospital, Montreal, Quebec, Canada. .,Biomedical Engineering Institute, University of Montreal, Montreal, Quebec, Canada. .,Department of Ophthalmology, University of Montreal, Montreal, Quebec, Canada.
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10
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Yellon SM, Mackler AM, Kirby MA. The Role of Leukocyte Traffic and Activation in Parturition. ACTA ACUST UNITED AC 2016. [DOI: 10.1016/s1071-55760300116-3] [Citation(s) in RCA: 37] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
Affiliation(s)
- S. M. Yellon
- Loma Linda University School of Medicine Center for Perinatal Biology, Departments of Physiologyand Anatomy, Loma Linda, California and Organon Pharmaceuticals, West Orange, New Jersey
| | | | - M. A. Kirby
- Loma Linda University School of Medicine Center for Perinatal Biology, Departments of Physiologyand Anatomy, Loma Linda, California and Organon Pharmaceuticals, West Orange, New Jersey
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11
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Nogueira-Neto J, Cardoso ASC, Monteiro HP, Fonseca FLA, Ramos LR, Junqueira VBC, Simon KA. Basal neutrophil function in human aging: Implications in endothelial cell adhesion. Cell Biol Int 2016; 40:796-802. [PMID: 27109745 DOI: 10.1002/cbin.10618] [Citation(s) in RCA: 20] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/05/2016] [Accepted: 04/21/2016] [Indexed: 01/28/2023]
Abstract
Much attention has been drawn to the pro-inflammatory condition that accompanies aging. This study compared parameters from non-stimulated neutrophils, obtained from young (18-30 years old [y.o.]) and elderly (65-80 y.o.) human volunteers. Measured as an inflammatory marker, plasmatic concentration of hs-CRP was found higher in elderly individuals. Non-stimulated neutrophil production of ROS and NO was, respectively, 38 and 29% higher for the aged group. From the adhesion molecules evaluated, only CD11b expression was elevated in neutrophils from the aged group, whereas no differences were found for CD11a, CD18, or CD62. A 69% higher non-stimulated in vitro neutrophil/endothelial cell adhesion was observed for neutrophils isolated from elderly donors. Our results suggest that with aging, neutrophils may be constitutively producing more reactive species in closer proximity to endothelial cells of vessel walls, which may both contribute to vascular damage and reflect a neutrophil intracellular disrupted redox balance, altering neutrophil function in aging.
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Affiliation(s)
- Joes Nogueira-Neto
- Department of Biological Sciences, Federal University of São Paulo, Diadema, SP, Brazil
| | - André S C Cardoso
- Department of Biological Sciences, Federal University of São Paulo, Diadema, SP, Brazil
| | - Hugo P Monteiro
- Department of Biochemistry/Molecular Biology, Federal University of São Paulo, São Paulo, SP, Brazil
| | - Fernando L A Fonseca
- Department of Biological Sciences, Federal University of São Paulo, Diadema, SP, Brazil.,Department of Hematology and Oncology, ABC School of Medicine, Santo André, SP, Brazil
| | - Luiz Roberto Ramos
- Department of Preventive Medicine, Federal University of São Paulo, São Paulo, SP, Brazil
| | | | - Karin A Simon
- Department of Biological Sciences, Federal University of São Paulo, Diadema, SP, Brazil.,Laboratório de Análises Clínicas e Toxicológicas, Instituto de Ciências Ambientais, Químicas e Farmacêuticas, Universidade Federal de São Paulo, Rua São Nicolau, 210-1° Andar, Diadema, 09913-030, SP, Brazil
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12
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Roos D. Complement and phagocytes - A complicated interaction. Mol Immunol 2016; 68:31-4. [PMID: 26597203 DOI: 10.1016/j.molimm.2015.05.001] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/27/2015] [Accepted: 05/01/2015] [Indexed: 01/11/2023]
Abstract
Mohamed Daha and I share a common interest in innate immunity. Working in institutes only 25 miles away from each other, that meant ample opportunity and relevance for collaboration. And so we did. Moreover, we have both been members of boards and councils of Dutch national organizations, and we have also become good friends. In this short recollection, I look back on 40 years of common activities in complement research and friendship.
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Affiliation(s)
- Dirk Roos
- Sanquin Research and University of Amsterdam, Amsterdam, The Netherlands.
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13
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Moussavi-Harami SF, Pezzi HM, Huttenlocher A, Beebe DJ. Simple microfluidic device for studying chemotaxis in response to dual gradients. Biomed Microdevices 2015; 17:9955. [PMID: 25893484 PMCID: PMC4768479 DOI: 10.1007/s10544-015-9955-8] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/23/2023]
Abstract
Chemotaxis is a fundamental biological process where complex chemotactic gradients are integrated and prioritized to guide cell migration toward specific locations. To understand the mechanisms of gradient dependent cell migration, it is important to develop in vitro models that recapitulate key attributes of the chemotactic cues present in vivo. Current in vitro tools for studying cell migration are not amenable to easily study the response of neutrophils to dual gradients. Many of these systems require external pumps and complex setups to establish and maintain the gradients. Here we report a simple yet innovative microfluidic device for studying cell migration in the presence of dual chemotactic gradients through a 3-dimensional substrate. The device is tested and validated by studying the migration of the neutrophil-like cell line PLB-985 to gradients of fMLP. Furthermore, the device is expanded and used with heparinised whole blood, whereupon neutrophils were observed to migrate from whole blood towards gradients of fMLP eliminating the need for any neutrophil purification or capture steps.
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Affiliation(s)
- S F Moussavi-Harami
- Department of Biomedical Engineering, University of Wisconsin, Madison, WI, USA
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14
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Lewis SM, Khan N, Beale R, Treacher DF, Brown KA. Depletion of blood neutrophils from patients with sepsis: treatment for the future? Int Immunopharmacol 2013; 17:1226-32. [PMID: 24144812 DOI: 10.1016/j.intimp.2013.10.002] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/30/2022]
Abstract
Organ failure arising from severe sepsis accounts for nearly 6 million deaths worldwide per annum. At present there are no specific pharmacological agents available for its treatment and identifying a suitable therapeutic target is urgently needed. Neutrophils appear to be contributing directly to pulmonary damage in severe forms of lung injury and indirectly to the failure of other organs. Blood neutrophils from patients with sepsis possess a phenotype that is indicative of activation and our results show that neutrophils isolated from patients with sepsis exhibit a supranormal adherence to endothelial monolayers treated with pro-inflammatory cytokines. Additional studies reveal that the patients' cells are highly efficient at releasing IL-8. We also demonstrate that organ function is improved upon removing neutrophils from the circulation. In this article we propose that in severe sepsis there is a subpopulation of neutrophils which is actively engaged in pathological insult. The phenotypic characterisation of this subset may provide a novel therapeutic strategy for sepsis that could lead to patient benefit.
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Affiliation(s)
- Sion M Lewis
- Intensive Care Unit, Guy's and St. Thomas' NHS Foundation Trust, London, UK; Vascular Immunology, King's College London, UK
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15
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van de Vijver E, van den Berg TK, Kuijpers TW. Leukocyte Adhesion Deficiencies. Hematol Oncol Clin North Am 2013; 27:101-16, viii. [DOI: 10.1016/j.hoc.2012.10.001] [Citation(s) in RCA: 54] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/23/2022]
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16
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Sharma J, Young DM, Marentette JO, Rastogi P, Turk J, McHowat J. Lung endothelial cell platelet-activating factor production and inflammatory cell adherence are increased in response to cigarette smoke component exposure. Am J Physiol Lung Cell Mol Physiol 2012; 302:L47-55. [PMID: 21984569 PMCID: PMC3349370 DOI: 10.1152/ajplung.00179.2011] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/06/2011] [Accepted: 10/03/2011] [Indexed: 12/30/2022] Open
Abstract
An early event in the pathogenesis of emphysema is the development of inflammation associated with accumulation of polymorphonuclear leukocytes (PMN) in small airways, and inflammatory cell recruitment from the circulation involves migration across endothelial and epithelial cell barriers. Platelet-activating factor (PAF) promotes transendothelial migration in several vascular beds, and we postulated that increased PAF production in the airways of smokers might enhance inflammatory cell recruitment and exacerbate inflammation. To examine this possibility, we incubated human lung microvascular endothelial cells (HMVEC-L) with cigarette smoke extract (CSE) and found that CSE inhibits PAF-acetylhydrolase (PAF-AH) activity. This enhances HMVEC-L PAF production and PMN adherence, and adherence is blocked by PAF receptor antagonists (CV3988 or ginkgolide B). CSE also inhibited PAF-AH activity of lung endothelial cells isolated from wild-type (WT) and iPLA(2)β knockout mice, and with WT cells, CSE enhanced PAF production and RAW 264.7 cell adherence. In contrast, CSE did not affect PAF production or RAW 264.7 cell adherence to iPLA(2)β-null cells, suggesting that iPLA(2)β plays an important role in PAF production by lung endothelial cells. These findings suggest that inhibition of PAF-AH by components of cigarette smoke may initiate or exacerbate inflammatory lung disease by enhancing PAF production and promoting accumulation of inflammatory cells in small airways. In addition, iPLA(2)β is identified as a potential target for therapeutic interventions to reduce airway inflammation and the progression of chronic lung disease.
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Affiliation(s)
- Janhavi Sharma
- Department of Pathology, Saint Louis University School of Medicine, St. Louis, Missouri 63104, USA
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17
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Kaposi's sarcoma-associated herpesvirus-encoded latency-associated nuclear antigen reduces interleukin-8 expression in endothelial cells and impairs neutrophil chemotaxis by degrading nuclear p65. J Virol 2011; 85:8606-15. [PMID: 21697472 DOI: 10.1128/jvi.00733-11] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/19/2022] Open
Abstract
Latency-associated nuclear antigen 1 (LANA-1) of Kaposi's sarcoma-associated herpesvirus (KSHV) is the major viral latent protein and functions as a multifaceted protein. Here, we report that LANA-1 attenuates the endothelial response to tumor necrosis factor alpha (TNF-α) stimulation and inhibits consequent neutrophil chemotaxis. Reporter assays showed that LANA-1 constantly repressed nuclear factor (NF)-κB transactivity upon TNF-α stimulation. We also found that LANA-1 decreased nuclear p65 protein levels through enhancement of polyubiquitylation-mediated p65 degradation and that an elongin B/elongin C-cullin 5-LANA-1-p65 complex assembled by LANA-1 was responsible for this enhanced p65 degradation. In telomerase-immortalized human umbilical vein endothelial cells, LANA-1 was demonstrated to repress interleukin-8 expression, which was involved in neutrophil recruitment to the inflammatory site. Through an in vitro transmigration assay, we determined a suppressive effect of LANA-1 on neutrophil chemotaxis. Our work suggests that KSHV LANA-1 is a negative modulator of acute inflammation and sheds light on a new mechanism by which KSHV during the latent life cycle evades the host innate immune response.
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18
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Ludwig A, Sommer A, Uhlig S. Assessment of endothelial permeability and leukocyte transmigration in human endothelial cell monolayers. Methods Mol Biol 2011; 763:319-32. [PMID: 21874462 DOI: 10.1007/978-1-61779-191-8_22] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
Increased vascular permeability is the hallmark of inflammation. Here, we describe three methods to assess vascular permeability in cell culture: (1) Impedance measurements of endothelial cell monolayers that allow to monitor changes in cell shape in real time. (2) Diffusion of fluorescently labeled dextran across endothelial cell monolayers to identify openings large enough for bulky molecules. (3) Transmigration of neutrophils through confluent endothelial cell monolayers to study one major process that increases endothelial permeability in inflammation.
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Affiliation(s)
- Andreas Ludwig
- Institute of Pharmacology and Toxicology, Faculty of Medicine, RWTH Aachen University, Aachen, Germany
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19
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Liangos O, Addabbo F, Tighiouart H, Goligorsky M, Jaber BL. Exploration of disease mechanism in acute kidney injury using a multiplex bead array assay: a nested case-control pilot study. Biomarkers 2010; 15:436-45. [PMID: 20482449 DOI: 10.3109/1354750x.2010.485252] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/08/2023]
Abstract
BACKGROUND Acute kidney injury (AKI) following cardiac surgery with cardiopulmonary bypass (CPB) causes increased morbidity and mortality. OBJECTIVE To evaluate the plasma profile of biomarkers potentially involved in AKI development following CPB. METHODS In a nested case-control study, plasma levels of 27 biomarkers in 11 AKI cases were compared with 25 controls. RESULTS Pre-CPB, plasma levels of epidermal growth factor and macrophage inflammatory protein-1beta, 2 h following CPB, soluble vascular cell adhesion molecule-1 (sVCAM-1), fractalkine and macrophage inflammatory protein-1alpha, and at later time points, sVCAM-1 and interleukin-6 were associated with AKI. CONCLUSION Biomarkers associated with AKI following CPB may merit further study.
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Affiliation(s)
- Orfeas Liangos
- Kidney & Dialysis Research Laboratory, St Elizabeth's Medical Center, Tufts University School of Medicine, Boston, MA, USA.
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20
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Abstract
Methods are described for analysing adhesion and migration of isolated lymphocytes on endothelial cell monolayers which have been co-cultured with different stromal cells, with or without additional cytokine treatment. The different cells types are grown on opposite sides of 3.0- or 0.4-mum pore filters depending on whether migration through the whole construct is to be analysed or adhesion to the endothelial cells alone. Assays may be "static" or filters can be incorporated into flow chambers so that cell behaviour can be directly observed under conditions simulating those in vivo. In general, by choice of method, one can evaluate efficiency of attachment and ability of cells to migrate across the endothelial monolayer, through the filter and through the stromal cell layer. Fluorescence microscopic examination of fixed filters can be used, e.g. to ascertain whether lymphocytes are retained by stromal cells. In general, static assays have the higher throughput and greatest ease of use, while the flow-based assays are more physiologically relevant and allow detailed recording of cell behaviour in real time.
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21
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Liangos O, Kolyada A, Tighiouart H, Perianayagam MC, Wald R, Jaber BL. Interleukin-8 and acute kidney injury following cardiopulmonary bypass: a prospective cohort study. Nephron Clin Pract 2009; 113:c148-54. [PMID: 19672112 DOI: 10.1159/000232595] [Citation(s) in RCA: 37] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/18/2008] [Accepted: 02/15/2009] [Indexed: 01/22/2023] Open
Abstract
BACKGROUND Cardiopulmonary bypass (CPB) elicits an inflammatory response mediated partly by neutrophils, which are activated and recruited by interleukin-8 (IL-8). We hypothesized that acute kidney injury (AKI) following CPB might be mediated by IL-8 and examined the association of perioperative plasma IL-8 levels with AKI in a prospective cohort. METHODS Plasma IL-8 was measured before, and 2, 24 and 48 h following CPB. Two AKI definitions, a serum creatinine increase of > or = 0.3 mg/dl or 50% (AKI Network [AKIN] stage-1) or > or = 50% alone (AKI-50%), within the first 72 h, were used. Area under the receiver operator characteristic curves (AUCs) were generated and multivariable logistic regression analyses performed. RESULTS A total of 143 patients were enrolled. The baseline mean serum creatinine was 1.1 mg/ dl (SD = 0.3), the CPB perfusion time was 112 min (SD = 43). Twenty-nine percent of the patients developed AKIN stage-1 and 13% AKI-50%. The plasma IL-8 level 2 h after CPB was higher in AKIN stage-1 (p = 0.03) and AKI-50% (p < 0.01), and predicted AKIN stage-1 (AUC = 0.62; p = 0.02) and AKI-50% (AUC = 0.72; p < 0.01). On multivariable analysis, the 2-hour plasma IL-8 level was associated with 1.36- and 1.59-fold higher odds for AKIN stage-1 and AKI-50%, respectively (p = 0.05). CONCLUSION Plasma IL-8 predicts the development of AKI following CPB, supporting a potential involvement for this chemokine in the pathogenesis of AKI.
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Affiliation(s)
- Orfeas Liangos
- Division of Nephrology, Caritas St. Elizabeth's Medical Center, Boston, MA 02135, USA.
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22
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Bauer S, Adrian N, Siebenborn U, Fadle N, Plesko M, Fischer E, Wüest T, Stenner F, Mertens JC, Knuth A, Ritter G, Old LJ, Renner C. Sequential cancer immunotherapy: targeted activity of dimeric TNF and IL-8. CANCER IMMUNITY 2009; 9:2. [PMID: 19267427 PMCID: PMC2935764] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Subscribe] [Scholar Register] [Received: 12/22/2008] [Accepted: 02/11/2009] [Indexed: 05/27/2023]
Abstract
Polymorphonuclear neutrophils (PMNs) are potent effectors of inflammation and their attempts to respond to cancer are suggested by their systemic, regional and intratumoral activation. We previously reported on the recruitment of CD11b+ leukocytes due to tumor site-specific enrichment of TNF activity after intravenous administration of a dimeric TNF immunokine with specificity for fibroblast activation protein (FAP). However, TNF-induced chemo-attraction and extravasation of PMNs from blood into the tumor is a multistep process essentially mediated by interleukin 8. With the aim to amplify the TNF-induced and IL-8-mediated chemotactic response, we generated immunocytokines by N-terminal fusion of a human anti-FAP scFv fragment with human IL-8 (IL-8(72)) and its N-terminally truncated form IL-8(3-72). Due to the dramatic difference in chemotaxis induction in vitro, we favored the mature chemokine fused to the anti-FAP scFv for further investigation in vivo. BALB/c nu/nu mice were simultaneously xenografted with FAP-positive or -negative tumors and extended chemo-attraction of PMNs was only detectable in FAP-expressing tissue after intravenous administration of the anti-FAP scFv-IL-8(72) construct. As TNF-activated PMNs are likewise producers and primary targets for IL-8, we investigated the therapeutic efficacy of co-administration of both effectors: Sequential application of scFv-IL-8(72) and dimeric IgG1-TNF fusion proteins significantly enhanced anti-tumor activity when compared either to a single effector treatment regimen or sequential application of non-targeted cytokines, indicating that the tumor-restricted sequential application of IL-8(72) and TNF is a promising approach for cancer therapy.
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Affiliation(s)
- Stefan Bauer
- Oncology Department, University Hospital Zurich, Zurich, Switzerland.
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23
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Simon SI, Sarantos MR, Green CE, Schaff UY. Leucocyte recruitment under fluid shear: mechanical and molecular regulation within the inflammatory synapse. Clin Exp Pharmacol Physiol 2008; 36:217-24. [PMID: 19018799 DOI: 10.1111/j.1440-1681.2008.05083.x] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/14/2023]
Abstract
1. Nature has evolved an exquisite system for regulation of leucocyte recruitment at sites of tissue inflammation. Mechanical energy translated to the red and white blood cells transports them from large arteries down to the microcirculation. 2. Neutrophils overcome the drag forces of blood flow by forming selectin and integrin adhesive bonds with the endothelium that coats the vessel wall. Leucocyte adhesion receptors have evolved unique mechanical and chemical properties that optimize for sequential binding and uptake of traction forces. 3. In the present brief review, we address how dispersive forces acting on a neutrophil in shear flow function to stabilize and synchronize bond formation within a macromolecular membrane complex we denote the inflammatory synapse.
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Affiliation(s)
- Scott I Simon
- Department of Biomedical Engineering, University of California, Davis, California 95616, USA.
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Rastogi P, White MC, Rickard A, McHowat J. Potential mechanism for recruitment and migration of CD133 positive cells to areas of vascular inflammation. Thromb Res 2008; 123:258-66. [PMID: 18495219 DOI: 10.1016/j.thromres.2008.03.020] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/21/2008] [Revised: 03/14/2008] [Accepted: 03/25/2008] [Indexed: 10/22/2022]
Abstract
OBJECTIVE Mast cells are found in large numbers in atherosclerotic plaques. The present study was conducted to determine whether tryptase stimulation of human coronary artery endothelial cells (HCAEC) would lead to an increase in transmigration of CD133 positive cells (CD133+). In vitro these cells can differentiate into mast cells under the influence of specific cytokines and growth factors. METHODS AND RESULTS CD133+ cells were isolated from umbilical cord blood. They express mRNA for several adhesion molecules that are also utilized in neutrophil migration and can migrate across an HCAEC monolayer. Migration increased significantly when HCAEC were stimulated with tryptase and decreased when CD133+ cells were pretreated with CV3988, a platelet activating factor receptor (PTAFR) antagonist. Following long-term cell culture, these cells stained positively for the presence of tryptase, a mast cell enzyme. CONCLUSION CD133+ cells can be utilized as a mast cell precursor population. The transendothelial migration is facilitated by the presence of tryptase and may utilize the PAF/PTAFR interaction in a manner similar to that involved in neutrophil transmigration. Following transmigration, a subset of these progenitor cells may mature into mast cells in the subendothelial space and play a role in propagation of the inflammatory process in atherosclerosis.
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Affiliation(s)
- Prerna Rastogi
- Department of Pathology, Saint Louis University School of Medicine, St. Louis, MO 63104, USA
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25
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Alam CAS, Seed MP, Freemantle C, Brown J, Perretti M, Carrier M, Divwedi A, West DC, Gustafson S, Colville-Nash PR, Willoughby DA. The inhibition of neutrophil-endothelial cell adhesion by hyaluronan independent of CD44. Inflammopharmacology 2007; 12:535-50. [PMID: 16259720 DOI: 10.1163/156856005774382733] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/19/2022]
Abstract
OBJECTIVE To study the effect of hyaluronan on cell adhesion and recruitment both in vitro and in vivo, since hyaluronan both inhibits restenosis and is anti-inflammatory. When administered to animals undergoing angioplasty the recruitment of cells into the restenotic plaque is inhibited, as well as into inflammatory lesions. The recent discovery that ICAM-1 binds hyaluronan and exhibits the B(X(7))B HA binding motif, led us also to investigate whether cell adhesion could be modulated by hyaluronan. MATERIALS AND METHODS Human neutrophils were adhered to human umbilical vein (HUVEC) or Ea.hy.926 HUVEC cells stimulated with phorbol myristate acetate (PMA) or tumour necrosis factor (TNFalpha). Neutrophil binding in vivo utilized FMLP-stimulated hamster cheek pouch post-capillary venules. RESULTS Hyaluronan inhibited human neutrophil adhesion to both PMA and TNFalpha-stimulated HUVEC. Ea.hy.926 human immortal HUVECs expressed ICAM-1 in response to TNFalpha and PMA. E-selectin was also upregulated by 6 h with TNFalpha but not significantly with PMA. TNFalpha induced CD44 expression within 4 h, but PMA not significantly up to 6 h. However, specific binding of [125I]hyaluronan to Ea.hy.926 cells was increased by PMA-stimulation at 4 h. Neutrophil adhesion to PMA-stimulated Ea.hy.926 HUVECs was inhibited in a concentration dependent fashion by both anti-ICAM-1 and hyaluronan (1 ng/ml-10 microg/ml) at 4 h. At 1 mg/ml adhesion was stimulated by hyaluronan. Hyaluronan had no effect on neutrophil adhesion to resting Ea.hy.926 cells. Hyaluronan (25 mg/kg, i.v.) inhibited cell adhesion to FMLP-stimulated post capillary venules of the hamster cheek pouch, whilst leaving cell rolling unaffected. CONCLUSIONS These results show that hyaluronan, at concentrations below those where intra-molecular associations occur, binds selectively to stimulated endothelial cells and inhibits neutrophil adhesion in vitro and in vivo via a mechanism which may involve molecules other than CD44, such as ICAM-1.
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Affiliation(s)
- C A S Alam
- Experimental Pathology Group, Biochemical Pharmacology, William Harvey Research Institute, Saint Bartholomew's & Royal London School of Medicine & Dentistry, Queen Mary & Westfield College, Charterhouse Square, London EC1M 6BQ, UK
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Abstract
In recent years, transfusion-related acute lung injury (TRALI) has developed from an almost unknown transfusion reaction to the most common cause of transfusion-related major morbidities and fatalities. A clinical definition of TRALI was established in 2004, based on acute respiratory distress, non-cardiogenic lung oedema temporal association with transfusion and hypoxaemia. Histological findings reveal lung oedema, capillary leucostasis and neutrophil extravasation. However, the pathogenesis of TRALI remains controversial. Leucocyte antibodies, present in fresh frozen plasma and platelet concentrates from multiparous donors, and neutrophil priming agents released in stored cellular blood components have been considered to be causative. As neutrophils and endothelial cells are pivotal in the pathogenesis of TRALI, a threshold model was established to try to unify the various reported findings on pathogenesis. This model comprises the priming of neutrophils and/or endothelium by the patient's co-morbidity, neutrophil and/or endothelial cell activation by the transfused blood component, and the severity of the TRALI reaction.
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Affiliation(s)
- Jürgen Bux
- DRK-Blood Service West of the German Red Cross, Hagen, Germany.
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27
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McGettrick HM, Butler LM, Nash GB. Analysis of leukocyte migration through monolayers of cultured endothelial cells. Methods Mol Biol 2007; 370:37-54. [PMID: 17416986 DOI: 10.1007/978-1-59745-353-0_4] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/14/2023]
Abstract
In this chapter methods are described for analyzing the adhesion and migration of isolated leukocytes on endothelial cell monolayers that have been cultured on different substrates and treated with cytokines. When endothelial cells are grown on porous filters inserted in wells, the levels of leukocyte adhesion and migration are calculated from the number added and the numbers retrieved from the upper and lower chambers. Fluorescence microscopic examination of the fixed filters can be used to ascertain whether leukocytes are retained above or below the filter. Direct observations of the time course of migration can be made when endothelial cells are cultured in six-well plates after leukocytes are allowed to settle onto them for a short period. In a more specialized assay, leukocytes are perfused through glass capillaries coated with endothelial cells, and again, direct video-microscopic observations are made. In this assay all stages of capture, immobilization, and migration can be followed. In general, the filter-based assay has the highest throughput and greatest ease of use but yields less detailed information, whereas the flow-based assay is most difficult to set up but is most physiologically relevant.
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Affiliation(s)
- Helen M McGettrick
- Division of Medical Sciences, Department of Physiology, The Medical School, , The University of Birmingham, Birmingham, UK
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28
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White MC, McHowat J. Protease activation of calcium-independent phospholipase A2 leads to neutrophil recruitment to coronary artery endothelial cells. Thromb Res 2006; 120:597-605. [PMID: 17188740 PMCID: PMC2170458 DOI: 10.1016/j.thromres.2006.11.007] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/05/2006] [Revised: 10/18/2006] [Accepted: 11/14/2006] [Indexed: 11/28/2022]
Abstract
INTRODUCTION Thrombin or tryptase cleavage of protease-activated receptors (PAR) on human coronary artery endothelial cells (HCAEC) results in activation of a membrane-associated, calcium-independent phospholipase A2 (iPLA2) that selectively hydrolyzes plasmalogen phospholipids. Atherosclerotic plaque rupture results in a coronary ischemic event in which HCAEC in the ischemic area would be exposed to increased thrombin concentrations in addition to tryptase released by activated mast cells present in the plaque. MATERIALS AND METHODS HCAEC were stimulated with thrombin or tryptase in the absence or presence of bromoenol lactone (BEL), a selective iPLA2 inhibitor, and iPLA2 activation, accumulation of biologically active membrane phospholipid-derived metabolites, upregulation of cell surface P-selectin expression and neutrophil adherence were measured. RESULTS HCAEC exposed to thrombin or tryptase stimulation demonstrated an increase in iPLA2 activity and arachidonic acid release. Additionally, stimulated HCAEC demonstrated increased platelet-activating factor (PAF) production and cell surface P-selectin expression, resulting in increased adhesion of neutrophils to HCAEC monolayers. Pretreatment with bromoenol lactone to inhibit iPLA2, blocked membrane phospholipid-derived metabolite production, increased cell surface P-selectin expression and neutrophil adherence. CONCLUSIONS The similar biochemical and cellular responses in HCAEC exposed to thrombin or tryptase stimulation suggest that the cleavage of two separate PAR serve to extend the range of proteases to which the cells respond rather than resulting in separate intracellular events. This suggests that in conditions such as thrombosis and atherosclerosis that multiple mechanisms can activate the inflammatory response.
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Affiliation(s)
- Maureen C White
- Saint Louis University School of Medicine, Department of Pathology, 1402 S. Grand, St. Louis, MO 63104, United States.
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30
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Sheikh S, Rahman M, Gale Z, Luu NT, Stone PCW, Matharu NM, Rainger GEL, Nash GB. Differing mechanisms of leukocyte recruitment and sensitivity to conditioning by shear stress for endothelial cells treated with tumour necrosis factor-alpha or interleukin-1beta. Br J Pharmacol 2005; 145:1052-61. [PMID: 15912126 PMCID: PMC1576231 DOI: 10.1038/sj.bjp.0706281] [Citation(s) in RCA: 37] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022] Open
Abstract
The cytokines tumour necrosis factor-alpha (TNFalpha) and interleukin-1beta (IL-1B) induce endothelial cells to recruit leukocytes. However, the exact adhesion and activation mechanisms induced by each cytokine, and their relative sensitivities to modulation by endothelial exposure to shear stress remain unclear. We cultured human umbilical vein endothelial cells (HUVEC) in glass capillaries at various shear stresses, with TNFalpha or IL-1B added for the last 4 h. Subsequently, human neutrophils were perfused over the HUVEC, and adhesion and migration were recorded. Both cytokines induced dose-dependent capture of neutrophils. However, while conditioning of HUVEC by increasing shear stress for 24 h diminished their response to TNFalpha, the response of HUVEC to IL-1B was similar at all shear stresses. The differing sensitivities were evident at levels of adhesive function and mRNA for adhesion molecules and chemokines. Analysis of nuclear factor kappaB (NF-kappaB)/Rel family of transcription factors showed that their expression and activation were modified by exposure to shear stress, but did not obviously explain differential responses to TNFalpha and IL-1B. Antibodies against selectins were effective against capture of neutrophils on TNFalpha-treated but not IL-1B-treated HUVEC. Stable adhesion was supported by beta2-integrins in each case. Activation of neutrophils occurred dominantly through CXC-chemokine receptor 2 (CXCR2) for TNFalpha-treated HUVEC, while blockade of CXCR1, CXCR2 and of platelet-activating factor receptors caused additive inhibition of migration on IL-1B-treated HUVEC. The mechanisms which underlie neutrophil recruitment, and their modulation by the haemodynamic environment, differ between cytokines. Interventions aimed against leukocyte recruitment may not operate equally in different inflammatory milieu.
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Affiliation(s)
- Sajila Sheikh
- Division of Medical Sciences, Department of Physiology, The Medical School, The University of Birmingham, Birmingham B15 2T
| | - Mahbub Rahman
- Division of Medical Sciences, Department of Physiology, The Medical School, The University of Birmingham, Birmingham B15 2T
| | - Zoe Gale
- Division of Medical Sciences, Department of Physiology, The Medical School, The University of Birmingham, Birmingham B15 2T
| | - N Thin Luu
- Division of Medical Sciences, Department of Physiology, The Medical School, The University of Birmingham, Birmingham B15 2T
| | - Philip C W Stone
- Division of Medical Sciences, Department of Physiology, The Medical School, The University of Birmingham, Birmingham B15 2T
| | - Nick M Matharu
- Division of Medical Sciences, Department of Physiology, The Medical School, The University of Birmingham, Birmingham B15 2T
| | - G Edward Luu Rainger
- Division of Medical Sciences, Department of Physiology, The Medical School, The University of Birmingham, Birmingham B15 2T
| | - Gerard B Nash
- Division of Medical Sciences, Department of Physiology, The Medical School, The University of Birmingham, Birmingham B15 2T
- Author for correspondence:
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Funahashi H, Okada Y, Sawai H, Takahashi H, Matsuo Y, Takeyama H, Manabe T. The role of glial cell line-derived neurotrophic factor (GDNF) and integrins for invasion and metastasis in human pancreatic cancer cells. J Surg Oncol 2005; 91:77-83. [PMID: 15999351 DOI: 10.1002/jso.20277] [Citation(s) in RCA: 36] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
BACKGROUND AND OBJECTIVES It is generally accepted that the malignancy of pancreatic cancer is dependent upon the extent of invasion as well as metastasis. However, the factors and mechanisms are incompletely understood. We investigated whether glial cell lined-derived neurotrophic factor (GDNF) enhances the invasive and adhesive behaviors of pancreatic cancer cells by altering of the expression of integrins. METHODS The expression of the GDNF receptor in pancreatic cancer cell lines (SW1990 and Capan-2) was confirmed by RT-PCR. Then we determined the expression of integrin subunits and the alteration of their expression by GDNF using flow-cytometric analysis and a cellular enzyme-linked immunosorbent assay (CELISA). Adhesion and invasion assay were performed to investigate whether increased integrin expression affected the interaction between cancer cells and ECM proteins. RESULTS The GDNF receptor subunits were expressed in pancreatic cancer cells. GDNF enhanced the expression of some of the integrin subunits and increased their adhesive and invasive abilities. The enhanced expression and associated increase in adhesive and invasive abilities were inhibited by blocking the GDNF receptor or the integrin beta1 subunit. CONCLUSION The enhancement of integrin expression by GDNF signaling through the GDNF receptor strongly influences invasion and adhesion to ECM proteins by pancreatic cancer cells.
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Affiliation(s)
- Hitoshi Funahashi
- Department of Gastroenterological Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
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Oka S, Sasada M, Yamamoto K, Nohgawa M, Takahashi A, Yamashita K, Yamada H, Uchiyama T. Nitric Oxide Derived from Human Umbilical Vein Endothelial Cells Inhibits Transendothelial Migration of Neutrophils. Int J Hematol 2005; 81:220-7. [PMID: 15814333 DOI: 10.1532/ijh97.04070] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
We evaluated the roles of nitric oxide (NO) derived from endothelial cells in neutrophil transendothelial migration (TEM). Pretreatment of human umbilical vein endothelial cells (HUVECs) with NG-nitro-L-arginine methyl ester hydrochloride (L-NAME) or NG-monomethyl L-arginine (L-NMMA), which are inhibitors of NO synthases, enhanced neutrophil TEM. Similar augmentation of TEM was observed in the presence of an NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy PTIO). Neutrophil TEM across L-NAME- or L-NMMA-treated HUVECs was inhibited by continuous NO supply by NO donors. These findings support the suggestion that continuous production of NO by endothelial cells suppresses neutrophil TEM. Flow cytometric analyses revealed that NO accumulates in neutrophils co-cultured with NO-producing HUVECs. A decreased amount of NO was detected in neutrophils co-cultured with L-NAME-treated HUVECs compared with neutrophils co-cultured with untreated HUVECs. Soluble guanylyl cyclase (sGC) is known as one of the most important targets of NO in neutrophils. 3-(53-Hydroxymethyl-23furyl)-1-benzyl indazole (YC-1), an activator of sGC, inhibited L-NAME-induced neutrophil TEM. It was interesting that inhibition of neutrophil sGC with 1-H[1,2,4-]oxadiazolo[4,3-a]quinoxalin-1-1 (ODQ) was sufficient to enhance TEM. These results suggest that NO derived from HUVECs acts on neutrophils to inhibit TEM, at least in part by activating sGC. Our findings imply the role of NO constitutively generated by HUVECs in protection against excessive neutrophil extravasation and unnecessary tissue damage under physiological conditions.
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Affiliation(s)
- Satoshi Oka
- Department of Hematology and Oncology, Graduate School of Medicine, Kyoto University, Japan
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Affiliation(s)
- L A Lasky
- Department of Immunology, Genentech Inc, 460 Point San Bruno Boulevard, South San Francisco, California 94080, USA
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Rosseau S, Wiechmann K, Moderer S, Selhorst J, Mayer K, Krüll M, Hocke A, Slevogt H, Seeger W, Suttorp N, Seybold J, Lohmeyer J. Moraxella catarrhalis–Infected Alveolar Epithelium Induced Monocyte Recruitment and Oxidative Burst. Am J Respir Cell Mol Biol 2005; 32:157-66. [PMID: 15557018 DOI: 10.1165/rcmb.2004-0091oc] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/24/2022] Open
Abstract
The recruitment of monocytes appears to be a crucial factor for inflammatory lung disease. Alveolar epithelial cells contribute to monocyte influx into the lung, but their impact on monocyte inflammatory capacity is not entirely clear. We thus analyzed the modulation of monocyte oxidative burst by A549 and isolated human alveolar epithelial cells. Epithelial infection with Moraxella catarrhalis induced monocyte adhesion, transepithelial migration, and superoxide generation, whereas stimulation with lipopolysaccharide, tumor necrosis factor-alpha, interleukin-1beta, or interferon-gamma induced adhesion or transmigration, but failed to initiate monocyte burst. The effect of microbial challenge was mimicked by phorbol myristate acetate and inhibited by the protein kinase C inhibitor bisindoylmaleimide. Furthermore, evidence for a role of platelet-activating factor-signaling in monocytes is presented. Monocyte burst was neither induced by supernatant nor affected by fixation of A549 cells, excluding the contribution of epithelium-derived soluble factors but emphasizing the mandatory role of intercellular contact. The employment of blocking antibodies, however, denied a role for the adhesion molecules intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, or CD11b/CD18 and CD49d/CD29. In essence, infection of alveolar epithelial cells with M. catarrhalis might amplify the inflammatory capacity of invading monocytes eliciting their superoxide production. The epithelial response to this microbial challenge thus clearly differed from that to proinflammatory cytokines.
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Affiliation(s)
- Simone Rosseau
- Department of Internal Medicine and Infectious Diseases, Charité-Campus Mitte, Schumannstrasse 20/21, 10117 Berlin, Germany.
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35
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Fuhler GM, Knol GJ, Drayer AL, Vellenga E. Impaired interleukin-8- and GROα-induced phosphorylation of extracellular signal-regulated kinase result in decreased migration of neutrophils from patients with myelodysplasia. J Leukoc Biol 2004; 77:257-66. [PMID: 15561756 DOI: 10.1189/jlb.0504306] [Citation(s) in RCA: 39] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Abstract
Patients with myelodysplasia suffer from recurrent bacterial infections as a result of differentiation defects of the myeloid lineage and a disturbed functioning of neutrophilic granulocytes. Important physiological activators of neutrophils are the cytokines interleukin-8/CXC chemokine ligand 8 (IL-8/CXCL8), which activates CXC chemokine receptor 1 and 2 (CXCR1 and CXCR2), and growth-related oncogene (GROalpha)/CXCL1, which stimulates only CXCR2. In this study, we show that migration toward IL-8/GROalpha gradients is decreased in myelodysplastic syndrome (MDS) neutrophils compared with healthy donors. We investigated the signal transduction pathways involved in IL-8/GROalpha-induced migration and showed that specific inhibitors for extracellular signal-regulated kinase (ERK)1/2 and phosphatidylinositol-3 kinase (PI-3K) abrogated neutrophil migration toward IL-8/GROalpha. In accordance with these results, we subsequently showed that IL-8/GROalpha-stimulated activation of ERK1/2 was substantially diminished in MDS neutrophils. Activation of the PI-3K downstream target protein kinase B/Akt was disturbed in MDS neutrophils when cells were activated with IL-8 but normal upon GROalpha stimulation. IL-8 stimulation resulted in higher migratory behavior and ERK1/2 activation than GROalpha stimulation, suggesting a greater importance of CXCR1. We then investigated IL-8-induced activation of the small GTPase Rac implicated in ERK1/2-dependent migration and found that it was less efficient in neutrophils from MDS patients compared with healthy donors. In contrast, IL-8 triggered a normal activation of the GTPases Ras and Ral, indicating that the observed defects were not a result of a general disturbance in CXCR1/2 signaling. In conclusion, our results demonstrate a disturbed CXCR1- and CXCR2-induced neutrophil chemotaxis in MDS patients, which might be the consequence of decreased Rac-ERK1/2 and PI-3K activation within these cells.
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Affiliation(s)
- Gwenny M Fuhler
- Division of Hematology, Department of Medicine, University Hospital Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands
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36
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Schornagel IJ, Sigurdsson V, Nijhuis EHJ, Bruijnzeel-Koomen CAFM, Knol EF. Decreased Neutrophil Skin Infiltration After UVB Exposure in Patients with Polymorphous Light Eruption. J Invest Dermatol 2004; 123:202-6. [PMID: 15191561 DOI: 10.1111/j.0022-202x.2004.22734.x] [Citation(s) in RCA: 40] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
Abstract
UV radiation, in particular UVB, suppresses the skin immune response. In patients with polymorphous light eruption (PLE) the skin immune response seems activated after UV exposure. Typical PLE skin lesions can occur as early as several hours after UV exposure. In healthy volunteers, neutrophils infiltrate the skin shortly after UV exposure. The kinetics and mechanisms of neutrophil infiltration in the skin of PLE patients after UVB exposure was studied. Skin biopsies at 0, 3, 6, and 18 h were taken from five PLE patients and six healthy controls after irradiation with three minimal erythema dose UVB. Furthermore, neutrophils were isolated from blood of five PLE patients and six healthy controls to test their chemotactic activity. Immunohistochemical analysis showed a significant decreased neutrophil infiltration in PLE skin after UVB irradiation compared with healthy controls (p<0.05). In both healthy controls and PLE patients, after UVB irradiation, ICAM-1 and E-selectin expression on endothelial cells increased at 6 h after irradiation. Blood neutrophil chemotactic response towards IL-8 and C5a, as well as the expression of cell surface markers involved in adhesion and chemotaxis, was not different between PLE patients and healthy controls. In conclusion, PLE is marked by a decreased skin infiltration of neutrophils after UVB irradiation, possibly leading to a diminished neutrophil-induced suppression.
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Affiliation(s)
- Ines J Schornagel
- Department of Dermatology/Allergology, University Medical Center Utrecht, Utrecht, The Netherlands.
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Kayem G, Maillard F, Batteux F, Weill B, Cabrol D, Goffinet F. Interleukin-8 mRNA in vaginal secretions: a prenatal marker of congenital infection in case of preterm labor with intact membranes. Prenat Diagn 2004; 24:58-62. [PMID: 14755411 DOI: 10.1002/pd.795] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/11/2022]
Abstract
OBJECTIVE To study whether interleukin-8 (IL-8) mRNA in vaginal secretions is associated with congenital infection and preterm delivery in the case of preterm labor with intact membranes. METHODS This prospective clinical study in a tertiary referral center included 280 patients who gave birth to 360 infants from 1997 through 1999. IL-8 mRNA in vaginal secretions was determined with reverse transcriptase polymerase chain reaction. Logistic regression was used to examine the association between vaginal IL-8 mRNA and congenital infection independently of the time of birth. Main outcome measures were congenital infection and delivery before 37 and 33 weeks' gestation. RESULTS A total of 100 women (100/280 (35.7%)) gave birth before 37 weeks. A total of 54 children (54/360 (15%)) had congenital infection. IL-8 mRNA in vaginal secretions was associated with delivery within 14 days of the sampling (24 (15.6%) vs. 7 (5.6%) p < 0.01), but not with delivery within 48 h, 7 days (p = 0.07) or before 37 or 33 weeks. There were more congenital infections in the group with detectable IL-8 mRNA (37 (19.3%) than in the negative group (17 (10.1%); p < 0.05). IL-8 mRNA was associated with congenital infection independently of the time of birth (OR: 2.6 (1.3-5.1)). This test had a sensitivity for predicting neonatal infection of 69%. Its specificity was 49%, its positive predictive value 19%, and its negative predictive value 90%. CONCLUSION IL-8 mRNA could be a prenatal noninvasive vaginal marker of congenital infection.
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Affiliation(s)
- G Kayem
- Department of Obstetrics and Gynaecology, Maternity Port-Royal, Cochin-Saint Vincent-de-Paul Hospital APHP, René Descartes University (Paris V), Paris, France.
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Hidemura A, Saito H, Fukatsu K, Ikeda S, Kitayama J, Matsuda T, Nagawa H. Patients with postoperative infections have sticky neutrophils before operation. Shock 2003; 19:497-502. [PMID: 12785002 DOI: 10.1097/01.sk.0000070733.34700.4e] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
Abstract
Appropriate polymorphonuclear neutrophil (PMN) recruitment is essential for host defense against infection. We investigated the significance of the preoperative PMN adhesion-migration process, as assessed by the flow chamber method, on postoperative infectious complications. Thirty-one consecutive patients with gastrointestinal malignancies, 21 colorectal and 10 gastric, who were undergoing elective surgery were enrolled. PMNs, isolated preoperatively from each patient's venous blood, were perfused onto a tumor necrosis factor alpha-stimulated human umbilical vein endothelial cell (HUVEC) monolayer through the flow chamber. We evaluated the adherent PMN number, the migrated PMN number, and the stuck PMN number by directly inspecting PMN interactions with a HUVEC monolayer under continuous shear flow simulating postcapillary venules. The expression of adhesion molecules on circulating PMNs was also measured. Patients were grouped into an infectious and a noninfectious group according to the occurrence of postoperative infectious complications defined by the Centers for Disease Control criteria. Eleven patients developed postoperative infectious complications. Although the number of preoperative in vitro adherent PMNs in patients with postoperative infection was significantly higher than in those without postoperative infection (P = 0.01), migrated PMN number was similar in both groups. Stuck PMN number tended to be higher in the infectious group than in the noninfectious group. The migrated PMN number showed a significant positive correlation with the adherent PMN number in the noninfectious group but not in the infectious group. Preoperative CD31 expression on circulating PMNs was significantly lower in the infectious group than in the noninfectious group. Preoperative in vitro derangement of the PMN adhesion-migration process is closely associated with postoperative infectious complications.
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Affiliation(s)
- Akio Hidemura
- Department of Surgery, Faculty of Medicine, University of Tokyo, Tokyo, Japan
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Sawai H, Takeyama H, Yamamoto M, Furuta A, Funahashi H, Okada Y, Sato M, Tanaka M, Manabe T. Enhancement of integrins by interleukin-1alpha, and their relationship with metastatic and invasive behavior of human pancreatic ductal adenocarcinoma cells. J Surg Oncol 2003; 82:51-6. [PMID: 12501168 DOI: 10.1002/jso.10187] [Citation(s) in RCA: 18] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
Abstract
BACKGROUND AND OBJECTIVES Adhesion and invasion of tumor cells to extracellular matrix (ECM) proteins play an important role in tumor metastasis formation. We investigated the enhancement of adhesive and invasive behavior to ECM proteins of human pancreatic cancer cells by interleukin-1alpha (IL-1alpha) to examine the mechanism of adhesion and invasion of metastatic human pancreatic cancer cells to ECM proteins. METHODS The enhancement of integrin subunits by IL-1alpha was examined by cellular enzyme-linked immunosorbent assay (CELISA) in two metastatic human pancreatic cancer cell lines (BxPC-3 and SW1990) and two nonmetastatic pancreatic cancer cell lines (PaCa-2 and PANC-1). In addition, assays of cancer cell adhesion and invasion to ECM proteins were performed to investigate whether increased integrin expression affected the invasive interaction between cancer cells and the putative integrin ECM ligands. RESULTS Expression of the alpha6 subunit by metastatic cancer cells was enhanced by IL-1alpha. Metastatic cancer cells also exhibited preferential adherence and invasion to laminin compared with nonmetastatic cancer cells, and this was enhanced by IL-1alpha. CONCLUSIONS The enhancement of alpha6beta1-integrin by Il-1alpha acting through IL-1RI, as well as the expression of alpha6beta1-integrin, plays an important role in metastasis formation in pancreatic cancer
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Affiliation(s)
- Hirozumi Sawai
- First Department of Surgery, Nagoya City University Medical School, Nagoya, Japan
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40
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Young RE, Thompson RD, Nourshargh S. Divergent mechanisms of action of the inflammatory cytokines interleukin 1-beta and tumour necrosis factor-alpha in mouse cremasteric venules. Br J Pharmacol 2002; 137:1237-46. [PMID: 12466233 PMCID: PMC1573606 DOI: 10.1038/sj.bjp.0704981] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022] Open
Abstract
1. Protein synthesis dependency and the role of endogenously generated platelet activating factor (PAF) and leukotriene B(4) (LTB(4)) in leukocyte migration through interleukin-1beta (IL-1beta)- and tumour necrosis factor-alpha (TNFalpha)-stimulated mouse cremasteric venules was investigated using established pharmacological interventions and the technique of intravital microscopy. 2. Based on previously obtained dose-response data, 30 ng rmIL-1beta and 300 ng rmTNFalpha were injected intrascrotally (4 h test period) to induce comparable levels of leukocyte firm adhesion and transmigration in mouse cremasteric venules. 3. Co-injection of the mRNA synthesis inhibitor, actinomycin D (0.2 mg kg(-1)), with the cytokines significantly inhibited firm adhesion (49+/-13.6%) and transmigration (67.2+/-4.2%) induced by IL-1beta, but not TNFalpha. 4. In vitro, TNFalpha (1-100 ng ml(-1)), but not IL-1beta, stimulated L-selectin shedding and increased beta(2) integrin expression on mouse neutrophils, as quantified by flow cytometry. 5. The PAF receptor antagonist, UK-74,505 (modipafant, 0.5 mg kg(-1), i.v.), had no effect on adhesion induced by either cytokine, but significantly inhibited transmigration induced by IL-1beta (66.5+/-4.5%). 6. The LTB(4) receptor antagonist, CP-105,696 (100 mg kg(-1), p.o.), significantly inhibited both IL-1beta induced adhesion (81.4+/-15.2%) and transmigration (58.7+/-7.2%), but had no effect on responses elicited by TNFalpha. Combined administration of the two antagonists had no enhanced inhibitory effects on responses induced by either cytokine. 7. The data indicate that firm adhesion and transmigration in mouse cremasteric venules stimulated by IL-1beta, but not TNFalpha, is protein synthesis dependent and mediated by endogenous generation of PAF and LTB(4). Additionally, TNFalpha but not IL-1beta, can directly stimulate mouse neutrophils in vitro. The findings provide further evidence to suggest divergent mechanisms of actions of IL-1beta and TNFalpha, two cytokines often considered to act via common molecular/cellular pathways.
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Affiliation(s)
- R E Young
- BHF Cardiovascular Medicine Unit, National Heart & Lung Institute, Faculty of Medicine, Imperial College London, Hammersmith Hospital, London W12 0NN, U.K
| | - R D Thompson
- BHF Cardiovascular Medicine Unit, National Heart & Lung Institute, Faculty of Medicine, Imperial College London, Hammersmith Hospital, London W12 0NN, U.K
| | - S Nourshargh
- BHF Cardiovascular Medicine Unit, National Heart & Lung Institute, Faculty of Medicine, Imperial College London, Hammersmith Hospital, London W12 0NN, U.K
- Author for correspondence:
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Rodriguez-Galán MC, Correa SG, Iribarren P, Sotomayor CE. Phenotypic and functional changes on phagocytic cells recruited at the site of Candida albicans infection after chronic varied stress exposure. Med Mycol 2002; 40:485-92. [PMID: 12462528 DOI: 10.1080/mmy.40.5.485.492] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/20/2022] Open
Abstract
The transition of Candida albicans from commensalism to pathogenicity is associated with the immune status of the host; resistance to fungus involves macrophages (Mphi) and polymorphonuclear neutrophils (PMN), which act as effector cells. T-cell function is also involved. Previously, we found that in Wistar rats exposed to chronic varied stress (CVS) immediately after C. albicans infection (Ca-S group) some functions of phagocytic cells, such as killer activity and NO production, were strongly modified compared with unstressed, infected animals (Ca group). We examined the phenotypic and functional changes of these effector cells recruited at the site of C. albicans infection. The recruitment of peritoneal cells (PC) was markedly reduced in Ca-S animals and the arrival of Mphi and PMN was selectively diminished after CVS exposure. The integrin CD11b/CD18, implicated in migration and C. albicans phagocytosis, was downregulated in Mphi of Ca-S animals. The activation markers CD54 and MHC-II were upregulated in Mphi after fungal contact. The expression of CD54 was only changed in Ca-S rats. Finally, TNF-alpha production was reduced in PC of Ca-S animals, suggesting an impairment of functional activity. Taken together, the phenotypic and functional changes detected in effector cells may account for the decreased resistance to candidiasis seen in conjunction with CVS. The changes seen also expand our knowledge of the role of Mphi in the control of C. albicans dissemination.
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Affiliation(s)
- M C Rodriguez-Galán
- Inmunología, Departamento de Bioquímica Clínica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Córdoba, Argentina
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Mayer K, Merfels M, Muhly-Reinholz M, Gokorsch S, Rosseau S, Lohmeyer J, Schwarzer N, Krüll M, Suttorp N, Grimminger F, Seeger W. Omega-3 fatty acids suppress monocyte adhesion to human endothelial cells: role of endothelial PAF generation. Am J Physiol Heart Circ Physiol 2002; 283:H811-8. [PMID: 12124231 DOI: 10.1152/ajpheart.00235.2002] [Citation(s) in RCA: 90] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
Abstract
Monocyte-endothelium interaction is a fundamental process in many acute and chronic inflammatory diseases. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are fish oil-derived alternative (omega-3) precursor fatty acids implicated in the suppression of inflammatory events. We investigated their influence on rolling and adhesion of monocytes to human umbilical vein endothelial cells (HUVEC) under laminar flow conditions in vitro. Exposure of HUVEC to tumor necrosis factor (TNF-alpha) strongly increased 1) surface expression of intercellular adhesion molecule (ICAM-1), vascular cell adhesion molecule (VCAM-1), and E-selectin, 2) platelet-activating factor (PAF) synthesis as assessed by thrombin challenge, and 3) rate of rolling and adhesion of monocytes. Preincubation of HUVEC with EPA or DHA markedly suppressed PAF synthesis, monocyte rolling, and adherence, whereas expression of endothelial adhesion molecules was unchanged. Also, PAF receptor antagonists markedly suppressed the adhesion rate of monocytes, and EPA or DHA revealed no additional inhibitory capacity. In contrast, arachidonic acid partially reversed the effect of the antagonist. We conclude that omega-3 fatty acids suppress rolling and adherence of monocytes on activated endothelial cells in vitro by affecting endothelial PAF generation.
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Affiliation(s)
- Konstantin Mayer
- Medizinische Klinik II, Justus Liebig University, Klinikstrasse 36, D-35392 Giessen, Germany.
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Cheng SS, Lukacs NW, Kunkel SL. Eotaxin/CCL11 is a negative regulator of neutrophil recruitment in a murine model of endotoxemia. Exp Mol Pathol 2002; 73:1-8. [PMID: 12127048 DOI: 10.1006/exmp.2002.2439] [Citation(s) in RCA: 18] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
Eotaxin/CCL11 is a chemokine that has been primarily characterized with respect to its eosinophil chemoattractant activity. However, the broad tissue expression of eotaxin/CCL11 suggests that it may have other unknown activities. We have used a murine model of endotoxemia to study the role of eotaxin/CCL11 in neutrophil recruitment. We demonstrate that eotaxin/CCL11 is acutely upregulated in the serum, peritoneal wash, and lungs of mice given an intraperitoneal lipopolysaccharide (LPS) challenge. Furthermore, immunoneutralization of eotaxin/CCL11 in this model results in a significant increase in the number of neutrophils within the lung after LPS challenge. When eotaxin/CCL11 knockout mice were challenged with LPS, these mice had increased peritoneal neutrophils, but not lung neutrophils, compared to the wild-type controls. Administration of eotaxin/CCL11 to eotaxin(-/-) mice suppressed endotoxemia-associated peritoneal neutrophils. The presence or absence of eotaxin/CCL11 did not affect the number of peritoneal macrophages in these mice. These data indicate that eotaxin/CCL11 plays a novel regulatory role during the acute inflammatory response and suggest that constitutive expression of this chemokine within tissues such as the gut, lung, heart, and placenta might be important in downregulating acute inflammatory processes within these tissues.
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Affiliation(s)
- Sara S Cheng
- Graduate Program in Cellular and Molecular Biology, University of Michigan Medical Center, Ann Arbor 48109, USA
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McClenahan DJ, Evanson OA, Weiss DJ. In vitro evaluation of the role of platelet-activating factor and interleukin-8 in Mannheimia haemolytica-induced bovine pulmonary endothelial cell injury. Am J Vet Res 2002; 63:394-401. [PMID: 11911574 DOI: 10.2460/ajvr.2002.63.394] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
OBJECTIVE To develop an in vitro model of the bovine alveolar-capillary interface and to evaluate the roles of interleukin-8 (IL-8) and platelet-activating factor (PAF) in neutrophil-mediated endothelial injury induced by infection with Mannheimia haemolytica. SAMPLE POPULATION Cultured bovine pulmonary microvascular endothelial cells, freshly isolated bovine neutrophils, and monocyte-derived bovine macrophages. PROCEDURE A coculture system was developed in which endothelial cells were grown to confluence in tissue culture inserts, neutrophils were added to the inserts, and macrophages were added to tissue culture wells. Mannheimia haemolytica-derived lipopolysaccharide (LPS) or supernatant was added to activate macrophages, and inhibitors of PAF or IL-8 were added to the insert. Endothelial cell cytotoxicity and permeability (ie, albumin leakage) and neutrophil activation (ie, adhesion, degranulation [lactoferrin expression], and superoxide production) were assessed. RESULTS The addition of M haemolytica-derived LPS to bovine macrophages in the coculture system resulted in significant increases in endothelial cell cytotoxicity and permeability and neutrophil degranulation and adhesion. Inhibition of IL-8 reduced endothelial cell permeability and neutrophil degranulation induced by exposure to M haemolytica-derived supernatant, whereas inhibition of PAF decreased superoxide release by neutrophils. CONCLUSIONS AND CLINICAL RELEVANCE In vitro activation of bovine macrophages by M haemolytica-derived LPS resulted in neutrophil activation and neutrophil-mediated endothelial damage. Neutrophil-mediated endothelial injury and neutrophil degranulation were, at least in part, mediated by IL8, whereas PAF promoted superoxide release by neutrophils in this in vitro system designed to mimic the in vivo events that occur during the early stages of bovine pneumonic pasteurellosis.
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Affiliation(s)
- David J McClenahan
- Department of Veterinary PathoBiology, College of Veterinary Medicine, University of Minnesota, St Paul 55108, USA
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Dobrina A, Pausa M, Fischetti F, Bulla R, Vecile E, Ferrero E, Mantovani A, Tedesco F. Cytolytically inactive terminal complement complex causes transendothelial migration of polymorphonuclear leukocytes in vitro and in vivo. Blood 2002; 99:185-92. [PMID: 11756170 DOI: 10.1182/blood.v99.1.185] [Citation(s) in RCA: 63] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
Intravital microscopy was used to monitor leukocyte traffic across rat mesenteric postcapillary venules induced by the inactive terminal complement (C) complex (iTCC) topically applied to ileal mesentery. Leukocytes started rolling within 15 minutes from the administration of iTCC, and by 1 hour they adhered almost completely to the endothelium emigrating from the vessels in the next 3 hours. C5a caused a similar, though less marked, effect, whereas boiled iTCC was inactive, excluding the contribution of contaminating lipopolysaccharide. The complex stimulated the migration of polymorphonuclear neutrophils (PMNs) across endothelial cells (ECs) in a transwell system after a 4-hour incubation of ECs with iTCC added to the lower chamber of the transwell, whereas a 30-minute incubation was sufficient for C5a and interleukin (IL)-8 to induce the passage of PMNs. C5a was not responsible for the effect of iTCC because this complex had no chemotactic activity and contained too small an amount of C5a to account for the transendothelial migration of PMNs. Similarly, the effect of iTCC was not mediated by IL-8 released by stimulated ECs because anti-IL-8 failed to inhibit the migration of PMNs induced by the complex. Unlike tumor necrosis factor-alpha, iTCC did not cause the redistribution of platelet-endothelial cell adhesion molecule-1 (PECAM-1), and PMN mobilization was partially blocked by anti-PECAM-1 antibodies.
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Affiliation(s)
- Aldo Dobrina
- Dipartimento di Fisiologia e Patologia, Università di Trieste, the IRCCS Burlo Garofolo, Trieste, Italy
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Sawai H, Yamamoto M, Okada Y, Sato M, Akamo Y, Takeyama H, Manabe T. Alteration of integrins by interleukin-1alpha in human pancreatic cancer cells. Pancreas 2001; 23:399-405. [PMID: 11668210 DOI: 10.1097/00006676-200111000-00011] [Citation(s) in RCA: 30] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/10/2022]
Abstract
INTRODUCTION Adhesion of tumor cells to extracellular matrix (ECM) proteins plays an important role in tumor invasion and metastasis. AIMS To investigate the expression of integrins in human pancreatic cancer cell lines and its alteration by interleukin (IL)-1alpha to examine the mechanism of adhesion of metastatic human pancreatic cancer cells to ECM proteins. METHODOLOGY The expression of integrin subunits and their alteration by IL-1alpha were examined by flow-cytometric analysis and cellular enzyme-linked immunosorbent assay in three metastatic human pancreatic cancer cell lines (AsPC-1, BxPC-3, and SW1990) and two nonmetastatic cancer cell lines (PaCa-2 and PANC-1). In addition, assays of cancer cell adhesion to ECM proteins were performed to investigate if increased integrin expression actually affected the adhesive interaction between cancer cells and the putative integrin ECM ligands. RESULTS The alpha(6) subunit expressed in metastatic cancer cells was enhanced by IL-1alpha. Metastatic cancer cells also showed preferential adherence to laminin compared with nonmetastatic cancer cells, and this was enhanced by IL-1alpha. CONCLUSION In pancreatic cancer, the enhancement of alpha(6)beta(1) integrin by IL-1alpha through IL-1 receptor type I, as well as the expression of alpha(6)beta(1) integrin, plays an important role in metastasis formation.
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Affiliation(s)
- H Sawai
- First Department of Surgery, Nagoya City University Medical School, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 4678601, Japan
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Zhang XW, Wang Y, Liu Q, Thorlacius H. Redundant function of macrophage inflammatory protein-2 and KC in tumor necrosis factor-alpha-induced extravasation of neutrophils in vivo. Eur J Pharmacol 2001; 427:277-83. [PMID: 11567658 DOI: 10.1016/s0014-2999(01)01235-3] [Citation(s) in RCA: 34] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
Abstract
Tumor necrosis factor-alpha (TNF-alpha) stimulates the expression CXC chemokines, i.e. macrophage inflammatory protein-2 (MIP-2) and cytokine-induced neutrophil chemoattractant (KC), and neutrophil extravasation. However, the individual role of MIP-2 and KC in the recruitment process of neutrophils in vivo remains elusive. By use of intravital microscopy in the mouse cremaster muscle, we analyzed the effect of specific inhibition of CXC chemokines, alone and together, on TNF-alpha-induced leukocyte rolling, firm adhesion and recruitment. After stimulation with TNF-alpha, the mRNA levels of both MIP-2 and KC were increased. Notably, separate administration of antibodies directed against MIP-2 and KC had no effect on TNF-alpha-induced neutrophil extravasation. In contrast, combined injection of anti-MIP-2 and anti-KC antibodies markedly inhibited extravascular migration of neutrophils. Moreover, MIP-2 and KC dose-dependently increased neutrophil recruitment, however, no synergistic effect of combined stimulation with MIP-2 and KC on the neutrophil response was found. Taken together, these data suggest that MIP-2 and KC are functionally redundant in TNF-alpha-induced neutrophil accumulation and that neutralization of both MIP-2 and KC may be necessary in order to reduce accumulation of neutrophils in cytokine-activated tissues.
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MESH Headings
- Animals
- Antibodies, Monoclonal/pharmacology
- Blood Flow Velocity/drug effects
- Capillary Permeability/drug effects
- Chemokine CXCL2
- Chemokines/genetics
- Chemokines/immunology
- Chemokines/pharmacology
- Chemotactic Factors/genetics
- Chemotactic Factors/immunology
- Chemotactic Factors/pharmacology
- Chemotaxis, Leukocyte/drug effects
- Dose-Response Relationship, Drug
- Erythrocytes/drug effects
- Erythrocytes/physiology
- Gene Expression Regulation/drug effects
- Leukocyte Count
- Male
- Mice
- Mice, Inbred BALB C
- Muscle, Skeletal/blood supply
- Muscle, Skeletal/drug effects
- Muscle, Skeletal/metabolism
- Neutrophil Infiltration/drug effects
- Neutrophils/cytology
- Neutrophils/drug effects
- RNA, Messenger/drug effects
- RNA, Messenger/genetics
- RNA, Messenger/metabolism
- Rats
- Tumor Necrosis Factor-alpha/pharmacology
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Affiliation(s)
- X W Zhang
- Department of Surgery, Malmö University Hospital, Lund University, 20502 Malmö, Sweden
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Heidari M, Harp JA, Kehrli ME. Expression, purification, and in vitro biological activities of recombinant bovine granulocyte-colony stimulating factor. Vet Immunol Immunopathol 2001; 81:45-57. [PMID: 11498246 DOI: 10.1016/s0165-2427(01)00321-x] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/27/2022]
Abstract
Neutrophils are essential components of the innate immune system and they play a critical role in the defense of host against bacterial and fungal infections. The colony stimulating factors are a class of glycoproteins that are required for proliferation, differentiation, and functional activation of hematopoietic progenitor cells. Granulocyte-colony stimulating factor (G-CSF) is a member of this regulatory family of cytokines that specifically stimulates proliferation and maturation of precursor cells in the bone marrow into fully differentiated and functional neutrophils. G-CSF also modulates the biological activities of mature neutrophils in circulation. A bovine G-CSF (bG-CSF) cDNA clone (previously isolated and sequenced in our laboratory) was expressed in Escherichia coli and the biological activities of the solubilized protein from purified inclusion bodies were examined. Flow cytometric analysis of membrane antigen density of neutrophils activated with bG-CSF revealed an upregulation in the expression of CD11a (>114%), CD11b (>148%), CD11c (>87%), and CD18 (>109%). Expression of L-selectin was decreased by more than 43%. There was no change, however, in the expression of CD14. These findings indicate that recombinant bG-CSF (rbG-CSF) expressed in E. coli is biologically active and exerts the same type of effects on neutrophils in vitro as those of human G-CSF (hG-CSF).
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Affiliation(s)
- M Heidari
- Periparturient Diseases of Cattle Research Unit, National Animal Disease Center, USDA-ARS, Ames, IA 50010, USA.
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Imai M, Tani A, Saito M, Saito K, Amano K, Nisijima M. Significance of fetal fibronectin and cytokine measurement in the cervicovaginal secretions of women at term in predicting term labor and post-term pregnancy. Eur J Obstet Gynecol Reprod Biol 2001; 97:53-8. [PMID: 11435010 DOI: 10.1016/s0301-2115(00)00483-8] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
Abstract
OBJECTIVE To determine whether fetal fibronectin (FFN) or cytokine concentrations in cervicovaginal secretions can be used to predict term labor and post-term pregnancy. STUDY DESIGN FFN and cytokines were assayed in cervicovaginal mucus from 122 pregnant women at 29-35 weeks and weekly from week 36 to parturition. RESULTS FFN concentrations were elevated from about 3 weeks before parturition; a correlation was found between FFN levels and sampling-to-delivery intervals. Parturition was best predicted within 7 days of sampling when the FFN value was >or=50ng/ml between 36 and 41 gestational weeks. Interleukin-1beta (IL-1beta) concentrations were elevated from 3 to 4 weeks before parturition; a correlation was found between IL-1beta levels and sampling-to-delivery intervals. Parturition was best predicted within 7 days of sampling, with an IL-1beta cut-off value of >or=100pg/ml. CONCLUSION Term labor and post-term pregnancy can be predicted within 7 days of sampling, using FFN and IL-1beta concentrations in cervicovaginal secretions of pregnant women.
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Affiliation(s)
- M Imai
- Department of Obstetrics and Gynecology, Kitasato University School of Medicine, 1-15-1, Kitasato Sagamihara, Kanagawa, Japan
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Arnould T, Thibaut-Vercruyssen R, Bouaziz N, Dieu M, Remacle J, Michiels C. PGF(2alpha), a prostanoid released by endothelial cells activated by hypoxia, is a chemoattractant candidate for neutrophil recruitment. THE AMERICAN JOURNAL OF PATHOLOGY 2001; 159:345-57. [PMID: 11438482 PMCID: PMC1850417 DOI: 10.1016/s0002-9440(10)61701-4] [Citation(s) in RCA: 41] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
Abstract
Despite increasing evidence supporting the involvement of neutrophils in ischemic and postischemic damages, the mechanisms underlying the early recruitment of these cells are not completely understood. In this report, the effects of conditioned media from hypoxic endothelial cells on neutrophil chemotaxis were investigated by biochemical and morphological studies. We showed that conditioned media collected from several endothelial cell origins submitted to hypoxia as well as ischemic rat liver perfusion liquids have a chemotactic activity for neutrophils. The role of various chemoattractant molecules like HETEs, platelet-activating factor, and cytokines such as interleukin-8 and interleukin-1 was examined in the same model. Chemotactic peptide contribution was ruled out as boiled conditioned media still trigger chemotaxis. However, cell treatment with cyclooxygenase inhibitors, neutralization of PGF(2alpha) biological activity with polyclonal antibodies, and the neutrophil preincubation with a specific PGF(2alpha) antagonist, all dramatically inhibited neutrophil chemotaxis. A strong chemoattractant effect of pure exogenous PGF(2alpha) or of a synthetic analog was also observed. The major effect of PGF(2alpha) on neutrophil chemotaxis was confirmed ex vivo in a rat liver perfusion ischemic model. These results suggest that PGF(2alpha), a prostanoid abundantly released by the endothelium of hypoxic or ischemic tissues, is a chemoattractant molecule that might be involved in the early recruitment of neutrophils in ischemic organs.
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Affiliation(s)
- T Arnould
- Laboratory of Biochemistry and Cellular Biology, University of Namur, Namur, Belgium.
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