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1
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Akiyama Y, Harada K, Miyakawa J, Kreder KJ, O’Donnell MA, Daichi M, Katoh H, Hori M, Owari K, Futami K, Ishikawa S, Ushiku T, Kume H, Homma Y, Luo Y. Th1/17 polarization and potential treatment by an anti-interferon-γ DNA aptamer in Hunner-type interstitial cystitis. iScience 2023; 26:108262. [PMID: 38026177 PMCID: PMC10663743 DOI: 10.1016/j.isci.2023.108262] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/20/2023] [Revised: 07/03/2023] [Accepted: 10/17/2023] [Indexed: 12/01/2023] Open
Abstract
Hunner-type interstitial cystitis (HIC) is a rare, enigmatic inflammatory disease of the urinary bladder with no curative treatments. In this study, we aimed to characterize the unique cellular and immunological factors specifically involved in HIC by comparing with cystitis induced by Mycobacterium bovis bacillus Calmette-Guérin, which presents similar clinicopathological features to HIC. Here, we show that T helper 1/17 +polarized immune responses accompanied by prominent overexpression of interferon (IFN)-γ, enhanced cGAS-STING cytosolic DNA sensing pathway, and increased plasma cell infiltration are the characteristic inflammatory features in HIC bladder. Further, we developed a mouse anti-IFN-γ DNA aptamer and observed that the intravesical instillation of the aptamer significantly ameliorated bladder inflammation, pelvic pain and voiding dysfunction in a recently developed murine HIC model with little migration into the blood. Our study provides the plausible basis for the clinical translation of the anti-IFN-γ DNA aptamer in the treatment of human HIC.
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Affiliation(s)
- Yoshiyuki Akiyama
- Department of Urology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
- Department of Urology, University of Iowa, Iowa City, IA, USA
| | | | - Jimpei Miyakawa
- Department of Urology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
| | - Karl J. Kreder
- Department of Urology, University of Iowa, Iowa City, IA, USA
| | | | - Maeda Daichi
- Department of Molecular and Cellular Pathology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
| | - Hiroto Katoh
- Department of Preventive Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
| | | | | | | | - Shumpei Ishikawa
- Department of Preventive Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
| | - Tetsuo Ushiku
- Department of Pathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
| | - Haruki Kume
- Department of Urology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
| | - Yukio Homma
- Department of Urology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
- Department of Interstitial Cystitis Medicine, Faculty of Medicine, Kyorin University, Tokyo, Japan
| | - Yi Luo
- Department of Urology, University of Iowa, Iowa City, IA, USA
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2
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Tay C, Grundy L. Animal models of interstitial cystitis/bladder pain syndrome. Front Physiol 2023; 14:1232017. [PMID: 37731545 PMCID: PMC10507411 DOI: 10.3389/fphys.2023.1232017] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2023] [Accepted: 08/01/2023] [Indexed: 09/22/2023] Open
Abstract
Interstitial Cystitis/Bladder Pain Syndrome (IC/BPS) is a chronic disorder characterized by pelvic and/or bladder pain, along with lower urinary tract symptoms that have a significant impact on an individual's quality of life. The diverse range of symptoms and underlying causes in IC/BPS patients pose a significant challenge for effective disease management and the development of new and effective treatments. To facilitate the development of innovative therapies for IC/BPS, numerous preclinical animal models have been developed, each focusing on distinct pathophysiological components such as localized urothelial permeability or inflammation, psychological stress, autoimmunity, and central sensitization. However, since the precise etiopathophysiology of IC/BPS remains undefined, these animal models have primarily aimed to replicate the key clinical symptoms of bladder hypersensitivity and pain to enhance the translatability of potential therapeutics. Several animal models have now been characterized to mimic the major symptoms of IC/BPS, and significant progress has been made in refining these models to induce chronic symptomatology that more closely resembles the IC/BPS phenotype. Nevertheless, it's important to note that no single model can fully replicate all aspects of the human disease. When selecting an appropriate model for preclinical therapeutic evaluation, consideration must be given to the specific pathology believed to underlie the development of IC/BPS symptoms in a particular patient group, as well as the type and severity of the model, its duration, and the proposed intervention's mechanism of action. Therefore, it is likely that different models will continue to be necessary for preclinical drug development, depending on the unique etiology of IC/BPS being investigated.
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Affiliation(s)
- Cindy Tay
- Neurourology Research Group, College of Medicine and Public Health, Flinders Health and Medical Research Institute, Flinders University, Adelaide, SA, Australia
| | - Luke Grundy
- Neurourology Research Group, College of Medicine and Public Health, Flinders Health and Medical Research Institute, Flinders University, Adelaide, SA, Australia
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3
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Piunti A, Meghani K, Yu Y, Robertson AG, Podojil JR, McLaughlin KA, You Z, Fantini D, Chiang M, Luo Y, Wang L, Heyen N, Qian J, Miller SD, Shilatifard A, Meeks JJ. Immune activation is essential for the antitumor activity of EZH2 inhibition in urothelial carcinoma. SCIENCE ADVANCES 2022; 8:eabo8043. [PMID: 36197969 PMCID: PMC9534493 DOI: 10.1126/sciadv.abo8043] [Citation(s) in RCA: 17] [Impact Index Per Article: 5.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 02/27/2022] [Accepted: 08/17/2022] [Indexed: 05/31/2023]
Abstract
The long-term survival of patients with advanced urothelial carcinoma (UCa) is limited because of innate resistance to treatment. We identified elevated expression of the histone methyltransferase EZH2 as a hallmark of aggressive UCa and hypothesized that EZH2 inhibition, via a small-molecule catalytic inhibitor, might have antitumor effects in UCa. Here, in a carcinogen-induced mouse bladder cancer model, a reduction in tumor progression and an increase in immune infiltration upon EZH2 inhibition were observed. Treatment of mice with EZH2i causes an increase in MHC class II expression in the urothelium and can activate infiltrating T cells. Unexpectedly, we found that the lack of an intact adaptive immune system completely abolishes the antitumor effects induced by EZH2 catalytic inhibition. These findings show that immune evasion is the only important determinant for the efficacy of EZH2 catalytic inhibition treatment in a UCa model.
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Affiliation(s)
- Andrea Piunti
- Division of Hematology/Oncology, Department of Pediatrics, University of Chicago, Chicago, IL, USA
- University of Chicago Medicine Comprehensive Cancer Center, Chicago, IL, USA
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
| | - Khyati Meghani
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
- Department of Urology, Feinberg School of Medicine, Chicago, IL, USA
| | - Yanni Yu
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
- Department of Urology, Feinberg School of Medicine, Chicago, IL, USA
| | | | - Joseph R. Podojil
- Department of Microbiology and Immunology, Feinberg School of Medicine, Chicago, IL, USA
| | - Kimberly A. McLaughlin
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
- Department of Urology, Feinberg School of Medicine, Chicago, IL, USA
| | - Zonghao You
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
- Department of Urology, Feinberg School of Medicine, Chicago, IL, USA
| | - Damiano Fantini
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
- Department of Urology, Feinberg School of Medicine, Chicago, IL, USA
| | - MingYi Chiang
- Department of Microbiology and Immunology, Feinberg School of Medicine, Chicago, IL, USA
| | - Yi Luo
- Department of Urology, University of Iowa, Iowa City, IA, USA
| | - Lu Wang
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
| | - Nathan Heyen
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
- Department of Urology, Feinberg School of Medicine, Chicago, IL, USA
- Dxige Research Inc., Courtenay, BC, Canada
| | - Jun Qian
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
- Department of Urology, Feinberg School of Medicine, Chicago, IL, USA
- Jesse Brown VA Medical Center, Chicago, IL, USA
| | - Stephen D. Miller
- Department of Microbiology and Immunology, Feinberg School of Medicine, Chicago, IL, USA
| | - Ali Shilatifard
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
| | - Joshua J. Meeks
- Department of Biochemistry and Molecular Genetics, Simpson Querrey Institute for Epigenetics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
- Department of Urology, Feinberg School of Medicine, Chicago, IL, USA
- Jesse Brown VA Medical Center, Chicago, IL, USA
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4
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Wang Y, Zhang Y, Li PC, Guo J, Huo F, Yang J, Jia R, Wang J, Huang Q, Theodorescu D, Yu H, Yan C. Development of novel aptamer-based targeted chemotherapy for bladder cancer. Cancer Res 2022; 82:1128-1139. [DOI: 10.1158/0008-5472.can-21-2691] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/12/2021] [Revised: 12/08/2021] [Accepted: 01/18/2022] [Indexed: 12/24/2022]
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5
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Deets KA, Nichols Doyle R, Rauch I, Vance RE. Inflammasome activation leads to cDC1-independent cross-priming of CD8 T cells by epithelial cell-derived antigen. eLife 2021; 10:e72082. [PMID: 34939932 PMCID: PMC8719880 DOI: 10.7554/elife.72082] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/09/2021] [Accepted: 12/21/2021] [Indexed: 12/17/2022] Open
Abstract
The innate immune system detects pathogens and initiates adaptive immune responses. Inflammasomes are central components of the innate immune system, but whether inflammasomes provide sufficient signals to activate adaptive immunity is unclear. In intestinal epithelial cells (IECs), inflammasomes activate a lytic form of cell death called pyroptosis, leading to epithelial cell expulsion and the release of cytokines. Here, we employed a genetic system to show that simultaneous antigen expression and inflammasome activation specifically in IECs is sufficient to activate CD8+ T cells. By genetic elimination of direct T cell priming by IECs, we found that IEC-derived antigens were cross-presented to CD8+ T cells. However, cross-presentation of IEC-derived antigen to CD8+ T cells only partially depended on IEC pyroptosis. In the absence of inflammasome activation, cross-priming of CD8+ T cells required Batf3+ dendritic cells (conventional type one dendritic cells [cDC1]), whereas cross-priming in the presence of inflammasome activation required a Zbtb46+ but Batf3-independent cDC population. These data suggest the existence of parallel inflammasome-dependent and inflammasome-independent pathways for cross-presentation of IEC-derived antigens.
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Affiliation(s)
- Katherine A Deets
- Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California, BerkeleyBerkeleyUnited States
| | - Randilea Nichols Doyle
- Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California, BerkeleyBerkeleyUnited States
| | - Isabella Rauch
- Department of Molecular Microbiology and Immunology, Oregon Health and Science UniversityPortlandUnited States
| | - Russell E Vance
- Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California, BerkeleyBerkeleyUnited States
- Cancer Research Laboratory, University of California, BerkeleyBerkeleyUnited States
- Howard Hughes Medical Institute, University of California, BerkeleyBerkeleyUnited States
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6
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Wu J, Abraham SN. The Roles of T cells in Bladder Pathologies. Trends Immunol 2021; 42:248-260. [PMID: 33536141 PMCID: PMC7914211 DOI: 10.1016/j.it.2021.01.003] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/28/2020] [Revised: 01/04/2021] [Accepted: 01/06/2021] [Indexed: 12/26/2022]
Abstract
T lymphocytes play important roles in the skin and mucosal surfaces such as the gut and lung. Until recently the contributions of T cells to mammalian bladder immunity were largely unknown. With newer techniques, including single-cell RNA sequencing and reporter mice, an understanding is emerging of T cell roles in bladder diseases (bacterial infections, bladder cancer, chronic inflammation). In these pathologies, many bladder T cell responses can be harmful to the host through suboptimal clearance of bacteria or cancer cells, or by modulating autoinflammation. Recent findings suggest that T cell behavior might be influenced by resident T cell interactions with the bladder microbiota and other immunostimulants. Thus, regulating bladder T cell functions could emerge as a putative immunotherapy to treat some bladder diseases.
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Affiliation(s)
- Jianxuan Wu
- Department of Immunology, Duke University Medical Center, Durham, NC 27710, USA
| | - Soman N Abraham
- Department of Immunology, Duke University Medical Center, Durham, NC 27710, USA; Department of Pathology, Duke University Medical Center, Durham, NC 27710, USA; Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA; Program in Emerging Infectious Diseases, Duke-National University of Singapore, Singapore 169857, Singapore.
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7
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Rousseau M, O’Brien CJ, Antequera E, Zdimerova H, Cansever D, Canton T, Zychlinsky Scharff A, Ingersoll MA. Identification of Sex Differences in Tumor-Specific T Cell Infiltration in Bladder Tumor-Bearing Mice Treated with BCG Immunotherapy. Bladder Cancer 2020. [DOI: 10.3233/blc-200384] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
Abstract
BACKGROUND: Bladder cancer is the fourth most common cancer for men. However, women are often diagnosed with later stage disease and have poorer outcomes. Whether immune-based sex differences contribute to this discrepancy is unclear. In addition, models to investigate tumor-specific immunity in bladder cancer, in the context of tumor development or response to therapy, are lacking. OBJECTIVE: To address this specific unmet need, we incorporated a commonly used model antigen, ovalbumin, into two well-established models of bladder cancer; the orthotopic MB49 cell line model and the carcinogenic BBN bladder cancer model. METHOD: We tested the utility of these models to investigate tumor-specific immunity in the context of immunotherapy in both sexes. RESULTS: We found that BCG vaccination, prior to weekly BCG instillation does not impart an immune-specific benefit to tumor-bearing mice in the context of multiple BCG instillations. Furthermore, tumors developed in the testes in male mice, precluding the use of the MB49 model to directly investigate sex-based immune differences. In the BBN model, we observed that more tumor antigen-specific CD8+ T cells infiltrated male bladders compared to female bladders in the context of BCG immunotherapy whereas regulatory T cells had higher levels of the exhaustion marker PD-1 in female mice. CONCLUSIONS: We propose our modified BBN model will contribute to our understanding of how tumor-specific immunity arises in bladder cancer. Additionally, the BBN bladder cancer model may help to uncover sex differences in tumor-specific immunity, which would provide valuable information for the development of new treatments or combination therapies for bladder cancer in women and men.
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Affiliation(s)
- Matthieu Rousseau
- Department of Immunology, Institut Pasteur, Paris, France
- INSERM U1223, Paris, France
| | - Conan J.O. O’Brien
- Department of Immunology, Institut Pasteur, Paris, France
- INSERM U1223, Paris, France
| | - Eduardo Antequera
- Department of Immunology, Institut Pasteur, Paris, France
- INSERM U1223, Paris, France
| | - Hana Zdimerova
- Department of Immunology, Institut Pasteur, Paris, France
- INSERM U1223, Paris, France
| | - Dilay Cansever
- Department of Immunology, Institut Pasteur, Paris, France
- INSERM U1223, Paris, France
| | - Tracy Canton
- Department of Immunology, Institut Pasteur, Paris, France
- INSERM U1223, Paris, France
| | | | - Molly A. Ingersoll
- Department of Immunology, Institut Pasteur, Paris, France
- INSERM U1223, Paris, France
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8
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Akiyama Y, Yao JR, Kreder KJ, O'Donnell MA, Lutgendorf SK, Lyu D, Maeda D, Kume H, Homma Y, Luo Y. Autoimmunity to urothelial antigen causes bladder inflammation, pelvic pain, and voiding dysfunction: a novel animal model for Hunner-type interstitial cystitis. Am J Physiol Renal Physiol 2020; 320:F174-F182. [PMID: 33308017 DOI: 10.1152/ajprenal.00290.2020] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Abstract
Recent evidence revealed that Hunner-type interstitial cystitis (HIC) is a robust inflammatory disease potentially associated with enhanced immune responses and histologically characterized by epithelial denudation and lymphoplasmacytic infiltration with frequent clonal expansion of infiltrating B cells. To date, few animal models that reproduce the histological and clinical correlates of HIC have yet been established. In the present study, we aimed to develop a novel animal model for HIC via autoimmunity to the bladder urothelium using the transgenic mouse model (URO-OVA) that expresses the membrane form of the model antigen ovalbumin (OVA) as a self-antigen on the bladder urothelium. OVA-specific lymphocytes (splenocytes) were generated by immunization of C57BL/6 mice with OVA protein and injected intravenously into URO-OVA mice. The splenocytes from OVA-immunized C57BL/6 mice showed increased interferon (IFN)-γ production in response to OVA stimulation in vitro. URO-OVA mice adoptively transferred with OVA-primed splenocytes developed cystitis exhibiting histological chronic inflammatory changes such as remarkable mononuclear cell infiltration predominantly composed of T and B lymphocytes, increased vascularity, and mucosal hyperemia in the bladder at days 7-28 with a peak at day 21 tested. No systemic inflammation was found in cystitis-induced URO-OVA mice, nor was any inflammation found in wild-type C57BL/6 mice adoptively transferred with OVA-primed splenocytes. Along with bladder inflammation, URO-OVA mice demonstrated significantly increased pelvic nociceptive responses, voiding dysfunction, and upregulated mRNA expression levels for IFN-γ, tumor necrosis factor-α (TNF-α), and substance P precursor in the bladder. This model reproduces the histological and clinical features of human HIC, providing a novel model for HIC research.
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Affiliation(s)
| | - Jian-Rong Yao
- Department of Urology, University of Iowa, Iowa City, Iowa
| | - Karl J Kreder
- Department of Urology, University of Iowa, Iowa City, Iowa.,Department of Obstetrics and Gynecology, University of Iowa, Iowa City, Iowa
| | | | - Susan K Lutgendorf
- Department of Urology, University of Iowa, Iowa City, Iowa.,Department of Obstetrics and Gynecology, University of Iowa, Iowa City, Iowa.,Department of Psychological and Brain Sciences, University of Iowa, Iowa City, Iowa
| | - Dan Lyu
- Department of Anesthesia, University of Iowa, Iowa City, Iowa
| | - Daichi Maeda
- Department of Clinical Genomics, Graduate School of Medicine, Osaka University, Osaka, Japan
| | - Haruki Kume
- Department of Urology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
| | - Yukio Homma
- Japanese Red Cross Medical Center, Tokyo, Japan
| | - Yi Luo
- Department of Urology, University of Iowa, Iowa City, Iowa
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9
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Ligon MM, Wang C, DeJong EN, Schulz C, Bowdish DME, Mysorekar IU. Single cell and tissue-transcriptomic analysis of murine bladders reveals age- and TNFα-dependent but microbiota-independent tertiary lymphoid tissue formation. Mucosal Immunol 2020; 13:908-918. [PMID: 32366865 PMCID: PMC7572484 DOI: 10.1038/s41385-020-0290-x] [Citation(s) in RCA: 28] [Impact Index Per Article: 5.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/03/2019] [Revised: 03/02/2020] [Accepted: 03/31/2020] [Indexed: 02/04/2023]
Abstract
Aging has multifaceted effects on the immune system, but how aging affects tissue-specific immunity is not well-defined. Bladder diseases characterized by chronic inflammation are highly prevalent in older women, but mechanisms by which aging promotes these pathologies remain unknown. Tissue transcriptomics of unperturbed, young and aged bladders identified a highly altered immune landscape as a fundamental feature of the aging female bladder. Detailed mapping of immune cells using single cell RNA-sequencing revealed novel subsets of macrophages and dendritic cells and unique changes to the immune repertoire in the aged bladder. B and T cells are highly enriched in aged bladders and spontaneously form organized bladder tertiary lymphoid tissues (bTLTs). Naïve, activated, and germinal center B cells and IgA+ plasma cells are found within bTLT and associated with increased urinary IgA concentrations. bTLTs form with increasing age and their formation is dependent on TNFα. Microbiota are not required to form bTLT, as aged germfree mice also harbor them. Thus, bTLTs require age-dependent TNFα but are independent of the microbiota. Our results indicate that chronic, age-associated inflammation underlies a fundamental alteration to the bladder and establishes a resource for further investigation of the bladder immune system in homeostasis, aging, and disease.
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Affiliation(s)
- Marianne M. Ligon
- Department of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, MO, 63110, USA
| | - Caihong Wang
- Department of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, MO, 63110, USA
| | - Erica N. DeJong
- McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada
| | - Christian Schulz
- McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada
| | - Dawn M. E. Bowdish
- McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada
| | - Indira U. Mysorekar
- Department of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, MO, 63110, USA.,Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, 63110, USA.,To whom correspondence should be addressed: Indira U. Mysorekar, Ph.D., Washington University School of Medicine, Depts. of Obstetrics and Gynecology & Pathology and Immunology, 660 S. Euclid Ave., St. Louis, MO 63110, Phone: 314-747-1329, Fax: 314-747-1350,
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10
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Allouch A, Di Primio C, Paoletti A, Lê-Bury G, Subra F, Quercioli V, Nardacci R, David A, Saïdi H, Cereseto A, Ojcius DM, Montagnac G, Niedergang F, Pancino G, Saez-Cirion A, Piacentini M, Gougeon ML, Kroemer G, Perfettini JL. SUGT1 controls susceptibility to HIV-1 infection by stabilizing microtubule plus-ends. Cell Death Differ 2020; 27:3243-3257. [PMID: 32514048 DOI: 10.1038/s41418-020-0573-5] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/04/2019] [Revised: 05/27/2020] [Accepted: 05/29/2020] [Indexed: 01/15/2023] Open
Abstract
Understanding the viral-host cell interface during HIV-1 infection is a prerequisite for the development of innovative antiviral therapies. Here we show that the suppressor of G2 allele of skp1 (SUGT1) is a permissive factor for human immunodeficiency virus (HIV)-1 infection. Expression of SUGT1 increases in infected cells on human brain sections and in permissive host cells. We found that SUGT1 determines the permissiveness to infection of lymphocytes and macrophages by modulating the nuclear import of the viral genome. More importantly, SUGT1 stabilizes the microtubule plus-ends (+MTs) of host cells (through the modulation of microtubule acetylation and the formation of end-binding protein 1 (EB1) comets). This effect on microtubules favors HIV-1 retrograde trafficking and replication. SUGT1 depletion impairs the replication of HIV-1 patient primary isolates and mutant virus that is resistant to raltegravir antiretroviral agent. Altogether our results identify SUGT1 as a cellular factor involved in the post-entry steps of HIV-1 infection that may be targeted for new therapeutic approaches.
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Affiliation(s)
- Awatef Allouch
- Cell Death and Aging Team, Gustave Roussy Cancer Campus, F-94805, Villejuif, France.,Laboratory of Molecular Radiotherapy, INSERM U1030, Gustave Roussy Cancer Campus, F-94805, Villejuif, France.,Gustave Roussy Cancer Campus, F-94805, Villejuif, France.,Université Paris-Saclay, 114 Rue Edouard Vaillant, F-94805, Villejuif, France
| | - Cristina Di Primio
- Bio@SNS Laboratory, Scuola Normale Superiore, Piazza dei Cavalieri 7, 56126, Pisa, Italy
| | - Audrey Paoletti
- Cell Death and Aging Team, Gustave Roussy Cancer Campus, F-94805, Villejuif, France.,Laboratory of Molecular Radiotherapy, INSERM U1030, Gustave Roussy Cancer Campus, F-94805, Villejuif, France.,Gustave Roussy Cancer Campus, F-94805, Villejuif, France.,Université Paris-Saclay, 114 Rue Edouard Vaillant, F-94805, Villejuif, France
| | - Gabrielle Lê-Bury
- INSERM U1016, Institut Cochin, F-75013, Paris, France.,CNRS, UMR 8104, F-75013, Paris, France.,Université Paris Descartes, Université de Paris, F-75006, Paris, France
| | - Frédéric Subra
- CNRS UMR 8113 LBPA, Ecole Normale Supérieure de Cachan, 61 Avenue du Président Wilson, F-94230, Cachan, France
| | - Valentina Quercioli
- Bio@SNS Laboratory, Scuola Normale Superiore, Piazza dei Cavalieri 7, 56126, Pisa, Italy
| | - Roberta Nardacci
- National Institute for Infectious Diseases "Lazzaro Spallanzani", Via Portuense 292, I-00149, Rome, Italy
| | - Annie David
- Unité HIV, inflammation and Persistance, 28 Rue du Dr Roux, F-75015, Paris, France
| | - Héla Saïdi
- Antiviral Immunity, Biotherapy and Vaccine Unit, Institut Pasteur, 25 Rue du Dr Roux, F-75015, Paris, France
| | - Anna Cereseto
- Laboratory of Molecular Virology, Centre for Integrative Biology, University of Trento, Via Sommarive 9, Povo, I-38123, Trento, Italy
| | - David M Ojcius
- Department of Biomedical Sciences, Arthur Dugoni School of Dentistry, University of the Pacific, San Francisco, CA, 94103, USA.,Université de Paris, F-75013, Paris, France
| | | | - Florence Niedergang
- INSERM U1016, Institut Cochin, F-75013, Paris, France.,CNRS, UMR 8104, F-75013, Paris, France.,Université Paris Descartes, Université de Paris, F-75006, Paris, France
| | - Gianfranco Pancino
- Unité HIV, inflammation and Persistance, 28 Rue du Dr Roux, F-75015, Paris, France
| | - Asier Saez-Cirion
- Unité HIV, inflammation and Persistance, 28 Rue du Dr Roux, F-75015, Paris, France
| | - Mauro Piacentini
- National Institute for Infectious Diseases "Lazzaro Spallanzani", Via Portuense 292, I-00149, Rome, Italy.,Department of Biology, University of Rome "Tor Vergata", Via della Ricerca Scientifica 1, I-00173, Rome, Italy
| | - Marie-Lise Gougeon
- Antiviral Immunity, Biotherapy and Vaccine Unit, Institut Pasteur, 25 Rue du Dr Roux, F-75015, Paris, France
| | - Guido Kroemer
- Université Paris Descartes, Université de Paris, F-75006, Paris, France.,INSERM U848, Gustave Roussy Cancer Campus, F-94805, Villejuif, France.,Metabolomics Platform, Gustave Roussy Cancer Campus, F-94805, Villejuif, France.,Equipe 11 Labellisée Ligue Contre le Cancer, Centre de Recherche des Cordeliers, INSERM U1138, F-75006, Paris, France.,Pôle de Biologie, Hôpital Européen Georges-Pompidou, AP-HP, F-75015, Paris, France.,Department of Women's and Children's Health, Karolinska Institute, Karolinska University Hospital, S-17176, Stockholm, Sweden
| | - Jean-Luc Perfettini
- Cell Death and Aging Team, Gustave Roussy Cancer Campus, F-94805, Villejuif, France. .,Laboratory of Molecular Radiotherapy, INSERM U1030, Gustave Roussy Cancer Campus, F-94805, Villejuif, France. .,Gustave Roussy Cancer Campus, F-94805, Villejuif, France. .,Université Paris-Saclay, 114 Rue Edouard Vaillant, F-94805, Villejuif, France. .,Department of Biomedical Sciences, Arthur Dugoni School of Dentistry, University of the Pacific, San Francisco, CA, 94103, USA.
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11
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Akiyama Y, Luo Y, Hanno PM, Maeda D, Homma Y. Interstitial cystitis/bladder pain syndrome: The evolving landscape, animal models and future perspectives. Int J Urol 2020; 27:491-503. [PMID: 32246572 DOI: 10.1111/iju.14229] [Citation(s) in RCA: 91] [Impact Index Per Article: 18.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/28/2019] [Accepted: 02/24/2020] [Indexed: 12/30/2022]
Abstract
Interstitial cystitis/bladder pain syndrome is a debilitating condition of unknown etiology characterized by persistent pelvic pain with lower urinary tract symptoms and comprises a wide variety of potentially clinically useful phenotypes with different possible etiologies. Current clinicopathological and genomic evidence suggests that interstitial cystitis/bladder pain syndrome should be categorized by the presence or absence of Hunner lesions, rather than by clinical phenotyping based on symptomatology. The Hunner lesion subtype is a distinct inflammatory disease with proven bladder etiology characterized by epithelial denudation and enhanced immune responses frequently accompanied by clonal expansion of infiltrating B cells, with potential engagement of infection. Meanwhile, the non-Hunner lesion subtype is a non-inflammatory disorder with little evidence of bladder etiology. It is potentially associated with urothelial malfunction and neurophysiological dysfunction, and frequently presents with somatic and/or psychological symptoms, that commonly result in central nervous sensitization. Animal models of autoimmune cystitis and neurogenic sensitization might serve as disease models for the Hunner lesion and non-Hunner lesion subtypes, respectively. Here, we revisit the taxonomy of interstitial cystitis/bladder pain syndrome according to current research, and discuss its potential pathophysiology and representative animal models. Categorization of interstitial cystitis/bladder pain syndrome based on cystoscopy is mandatory to design optimized treatment and research strategies for each subtype. A tailored approach that specifically targets the characteristic inflammation and epithelial denudation for the Hunner lesion subtype, or the urothelial malfunction, sensitized/altered nervous system and psychosocial problems for the non-Hunner lesion subtype, is essential for better clinical management and research progress in this complex condition.
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Affiliation(s)
- Yoshiyuki Akiyama
- Department of Urology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.,Department of Urology, University of Iowa, Iowa City, Iowa, USA
| | - Yi Luo
- Department of Urology, University of Iowa, Iowa City, Iowa, USA
| | - Philip M Hanno
- Department of Urology, Stanford University School of Medicine, Stanford, California, USA
| | - Daichi Maeda
- Department of Clinical Genomics, Graduate School of Medicine, Osaka University, Osaka, Japan
| | - Yukio Homma
- Japanese Red Cross Medical Center, Tokyo, Japan
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12
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Update on the Pathophysiology of Interstitial Cystitis /Bladder Pain Syndrome. CURRENT BLADDER DYSFUNCTION REPORTS 2020. [DOI: 10.1007/s11884-019-00569-w] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/25/2022]
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13
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Cui X, Jing X, Lutgendorf SK, Bradley CS, Schrepf A, Erickson BA, Magnotta VA, Ness TJ, Kreder KJ, O'Donnell MA, Luo Y. Cystitis-induced bladder pain is Toll-like receptor 4 dependent in a transgenic autoimmune cystitis murine model: a MAPP Research Network animal study. Am J Physiol Renal Physiol 2019; 317:F90-F98. [PMID: 31091120 DOI: 10.1152/ajprenal.00017.2019] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/05/2023] Open
Abstract
Altered Toll-like receptor (TLR)4 activation has been identified in several chronic pain conditions but has not been well studied in interstitial cystitis/bladder pain syndrome (IC/BPS). Our previously published human studies indicated that patients with IC/BPS present altered systemic TLR4-mediated inflammatory responses, which were significantly correlated with reported pain severity. In the present study, we sought to determine whether altered TLR4 activation plays a role in pelvic/bladder pain seen in patients with IC/BPS using our validated IC/BPS-like transgenic autoimmune cystitis model (URO-OVA). URO-OVA mice developed responses consistent with pelvic and bladder pain after cystitis induction, which was associated with increased splenocyte production of TLR4-mediated proinflammatory cytokines IL-1β, IL-6, and TNF-α. Increased spinal expression of mRNAs for proinflammatory cytokines IL-6 and TNF-α, glial activation markers CD11b and glial fibrillary acidic protein, and endogenous TLR4 ligand high mobility group box 1 was also observed after cystitis induction. Compared with URO-OVA mice, TLR4-deficient URO-OVA mice developed significantly reduced nociceptive responses, although similar bladder inflammation and voiding dysfunction, after cystitis induction. Intravenous administration of TAK-242 (a TLR4-selective antagonist) significantly attenuated nociceptive responses in cystitis-induced URO-OVA mice, which was associated with reduced splenocyte production of TLR4-mediated IL-1β, IL-6, and TNF-α as well as reduced spinal expression of mRNAs for IL-6, TNF-α, CD11b, glial fibrillary acidic protein, and high mobility group box 1. Our results indicate that altered TLR4 activation plays a critical role in bladder nociception independent of inflammation and voiding dysfunction in the URO-OVA model, providing a potential mechanistic insight and therapeutic target for IC/BPS pain.
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Affiliation(s)
- Xiangrong Cui
- Department of Urology, University of Iowa , Iowa City, Iowa
| | - Xuan Jing
- Department of Urology, University of Iowa , Iowa City, Iowa
| | - Susan K Lutgendorf
- Department of Urology, University of Iowa , Iowa City, Iowa.,Department of Psychological and Brain Sciences, University of Iowa , Iowa City, Iowa.,Department of Obstetrics and Gynecology, University of Iowa , Iowa City, Iowa
| | - Catherine S Bradley
- Department of Urology, University of Iowa , Iowa City, Iowa.,Department of Obstetrics and Gynecology, University of Iowa , Iowa City, Iowa
| | - Andrew Schrepf
- Department of Psychological and Brain Sciences, University of Iowa , Iowa City, Iowa.,Department of Anesthesiology, University of Michigan , Ann Arbor, Michigan
| | | | | | - Timothy J Ness
- Department of Anesthesiology and Perioperative Medicine, University of Alabama at Birmingham , Birmingham, Alabama
| | - Karl J Kreder
- Department of Urology, University of Iowa , Iowa City, Iowa.,Department of Obstetrics and Gynecology, University of Iowa , Iowa City, Iowa
| | | | - Yi Luo
- Department of Urology, University of Iowa , Iowa City, Iowa
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14
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Zhang L, Ihsan AU, Cao Y, Khan FU, Cheng Y, Han L, Zhou X. An Immunogenic Peptide, T2 Induces Interstitial Cystitis/Painful Bladder Syndrome: an Autoimmune Mouse Model for Interstitial Cystitis/Painful Bladder Syndrome. Inflammation 2018; 40:2033-2041. [PMID: 28799018 DOI: 10.1007/s10753-017-0643-0] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
The exact pathophysiology of interstitial cystitis/painful bladder syndrome is unknown; however, autoimmunity is a valid theory. We developed an autoimmune chronic cystitis model by administration of the medium dose of immunogenic peptide T2. Sixty female C57BL/6 mice were divided into six groups. The control group was not treated with any reagent. CFA group was injected with CFA + normal saline, homogenate group with bladder homogenate + CFA, low-dose group with low dose of T2 peptide + CFA, medium dose group with the medium dose of T2 peptide + CFA, and high-dose group with the high dose of T2 peptide + CFA. Micturition habits, withdrawal frequencies of mice, and bladders weight were measured for each group. Hematoxylin and eosin staining and toluidine blue staining were used to investigate bladder inflammation and mast cells accumulation, respectively. T cells infiltration in the bladder tissues and serum TNF-α level were measured by using immunohistochemistry and ELISA, respectively. Mice immunized with the medium dose of T2 peptide (0.225 mg/ml) were extremely sensitive to the applied force, showed greater urine frequencies, and higher bladder weights. Histologic examination revealed severe edema and inflammation in bladder tissues of medium-dose group. Extensive infiltration of T cells in bladder tissues, elevated TNF-α, and increased mast cells accumulation were observed in medium-dose group as compared to that in other groups. EAC mice model established by injecting the medium dose of T2 (0.225 mg/ml) mimics all the symptoms and pathophysiologic characteristics of IC/PBS. We believe that this model can help us to investigate the pathogenesis of IC/PBS.
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Affiliation(s)
- Li Zhang
- Department of Clinical Pharmacy, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu province, 211198, People's Republic of China
| | - Awais Ullah Ihsan
- Department of Clinical Pharmacy, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu province, 211198, People's Republic of China
| | - Yanfang Cao
- Department of Clinical Pharmacy, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu province, 211198, People's Republic of China
| | - Farhan Ullah Khan
- Department of Clinical Pharmacy, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu province, 211198, People's Republic of China
| | - Yijie Cheng
- Department of Clinical Pharmacy, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu province, 211198, People's Republic of China
| | - Lei Han
- Department of Pharmacy, Jiangsu Jiankang Vocational College, Nanjing, Jiangsu province, 211198, People's Republic of China.,Department of Pharmacy, Jiangsu Worker Medical university, Nanjing, Jiangsu province, 211198, People's Republic of China
| | - Xiaohui Zhou
- Department of Clinical Pharmacy, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu province, 211198, People's Republic of China. .,Department of Surgery, Nanjing Shuiximen Hospital, Nanjing, Jiangsu province, 211198, People's Republic of China. .,Department of Surgery, Zhongda Hospital, Nanjing, Jiangsu province, 210009, People's Republic of China.
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15
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Sub-noxious Intravesical Lipopolysaccharide Triggers Bladder Inflammation and Symptom Onset in A Transgenic Autoimmune Cystitis Model: A MAPP Network Animal Study. Sci Rep 2018; 8:6573. [PMID: 29700406 PMCID: PMC5919907 DOI: 10.1038/s41598-018-24833-x] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/12/2018] [Accepted: 04/11/2018] [Indexed: 11/21/2022] Open
Abstract
Patients with interstitial cystitis/bladder pain syndrome (IC/BPS) can potentially develop symptom flares after exposure to minor bladder irritants such as subclinical bacterial infection. To reproduce this symptom onset, we intravesically instilled a sub-noxious dose of uropathogenic E. coli component lipopolysaccharide (LPS) in young URO-OVA/OT-I mice, a transgenic autoimmune cystitis model that spontaneously develops bladder inflammation at ≥10 weeks of age. Female URO-OVA/OT-I mice (6-weeks old) were treated intravesically with phosphate-buffered saline (PBS) or PBS containing a sub-noxious dose (1 μg) of LPS. Mice were evaluated for bladder inflammation, pelvic pain, and voiding dysfunction at days 1, 7, and 14 post-treatment. Mice treated with LPS but not PBS developed early bladder inflammation with increased macrophage infiltration. Accordingly, the inflamed bladders expressed increased levels of mRNA for proinflammatory cytokines (IL-1β and IL-6) and pain mediator (substance P precursor). In addition, LPS-treated mice exhibited pelvic pain and voiding dysfunction such as increased urinary frequency and reduced bladder capacity. These functional changes sustained up to day 14 tested. Our results indicate that a single sub-noxious dose of intravesical LPS triggers early bladder inflammation and symptom onset in URO-OVA/OT-I mice, providing a useful model for IC/BPS symptom flare study.
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16
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Moldenhauer LM, Diener KR, Hayball JD, Robertson SA. An immunogenic phenotype in paternal antigen-specific CD8 + T cells at embryo implantation elicits later fetal loss in mice. Immunol Cell Biol 2017; 95:705-715. [PMID: 28529323 DOI: 10.1038/icb.2017.41] [Citation(s) in RCA: 24] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/17/2017] [Revised: 05/16/2017] [Accepted: 05/16/2017] [Indexed: 02/07/2023]
Abstract
Central to pregnancy success is a state of T cell tolerance to paternal antigens, which is initiated at conception. The role and regulation of specific phenotypes of CD8+ T cells in mediating pregnancy tolerance is not clear. This study aimed to investigate the impact on pregnancy outcome of altering the cytokine environment during maternal CD8+ T cell priming in early pregnancy. Transgenic Act-mOVA male mice were mated to C57BL/6 (B6) females to generate fetuses expressing ovalbumin (OVA) as a model paternal antigen. OVA-reactive CD8+ OT-I T cells were activated in vitro with OVA in the presence of either transforming growth factor-β1 (TGFB1) plus interleukin-10 (IL10), or IL2, to mimic normal or dysregulated uterine conditions, respectively, and transferred into pregnant mice on gestational day 3.5. OT-I T cells activated with TGFB1 and IL10, like naive OT-I T cells, did not alter embryo implantation or fetal viability. In contrast, OT-I T cells activated with IL2 caused extensive fetal loss manifesting in mid-gestation. IL2-activated OT-I T cells expressed less FOXP3 and higher interferon-γ (IFNG) than cells activated with TGFB1 and IL10. Fetal loss did not occur in females mated with B6 males, demonstrating the antigen specificity of fetal loss, and was not abrogated by maternal genetic C1q deficiency indicating a mechanism independent of antibody-mediated cytotoxicity. These data indicate that alternative phenotypes generated in maternal CD8+ T cells at the time of priming with paternal antigens can impact pregnancy outcome, such that inappropriate activation of CD8+ T cells before implantation is capable of causing antigen-specific fetal loss later in pregnancy.
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Affiliation(s)
- Lachlan M Moldenhauer
- Robinson Research Institute and Adelaide Medical School, The University of Adelaide, Adelaide, South Australia, Australia
| | - Kerrilyn R Diener
- Robinson Research Institute and Adelaide Medical School, The University of Adelaide, Adelaide, South Australia, Australia.,School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, South Australia, Australia.,Experimental Therapeutics Laboratory, Hanson Institute and Sansom Institute for Health Research, Adelaide, South Australia, Australia
| | - John D Hayball
- Robinson Research Institute and Adelaide Medical School, The University of Adelaide, Adelaide, South Australia, Australia.,School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, South Australia, Australia.,Experimental Therapeutics Laboratory, Hanson Institute and Sansom Institute for Health Research, Adelaide, South Australia, Australia
| | - Sarah A Robertson
- Robinson Research Institute and Adelaide Medical School, The University of Adelaide, Adelaide, South Australia, Australia
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17
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Wang X, Liu W, O'Donnell M, Lutgendorf S, Bradley C, Schrepf A, Liu L, Kreder K, Luo Y. Evidence for the Role of Mast Cells in Cystitis-Associated Lower Urinary Tract Dysfunction: A Multidisciplinary Approach to the Study of Chronic Pelvic Pain Research Network Animal Model Study. PLoS One 2016; 11:e0168772. [PMID: 28002455 PMCID: PMC5176179 DOI: 10.1371/journal.pone.0168772] [Citation(s) in RCA: 32] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/11/2016] [Accepted: 11/16/2016] [Indexed: 11/19/2022] Open
Abstract
Bladder inflammation frequently causes cystitis pain and lower urinary tract dysfunction (LUTD) such as urinary frequency and urgency. Although mast cells have been identified to play a critical role in bladder inflammation and pain, the role of mast cells in cystitis-associated LUTD has not been demonstrated. Interstitial cystitis/bladder pain syndrome (IC/BPS) is a chronic and debilitating inflammatory condition of the urinary bladder characterized by the hallmark symptoms of pelvic pain and LUTD. In this study we investigated the role of mast cells in LUTD using a transgenic autoimmune cystitis model (URO-OVA) that reproduces many clinical correlates of IC/BPS. URO-OVA mice express the membrane form of the model antigen ovalbumin (OVA) as a self-antigen on the urothelium and develop bladder inflammation upon introduction of OVA-specific T cells. To investigate the role of mast cells, we crossed URO-OVA mice with mast cell-deficient KitW-sh mice to generate URO-OVA/KitW-sh mice that retained urothelial OVA expression but lacked endogenous mast cells. We compared URO-OVA mice with URO-OVA/KitW-sh mice with and without mast cell reconstitution in response to cystitis induction. URO-OVA mice developed profound bladder inflammation with increased mast cell counts and LUTD, including increased total number of voids, decreased mean volume voided per micturition, and decreased maximum volume voided per micturition, after cystitis induction. In contrast, similarly cystitis-induced URO-OVA/KitW-sh mice developed reduced bladder inflammation with no mast cells and LUTD detected. However, after mast cell reconstitution URO-OVA/KitW-sh mice restored the ability to develop bladder inflammation and LUTD following cystitis induction. We further treated URO-OVA mice with cromolyn, a mast cell membrane stabilizer, and found that cromolyn treatment reversed bladder inflammation and LUTD in the animal model. Our results provide direct evidence for the role of mast cells in cystitis-associated LUTD, supporting the use of mast cell inhibitors for treatment of certain forms of IC/BPS.
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Affiliation(s)
- Xu Wang
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
- Department of Obstetrics and Gynecology, University of Iowa, Iowa City, Iowa, United States of America
| | - Wujiang Liu
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
| | - Michael O'Donnell
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
| | - Susan Lutgendorf
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
- Tianjin Institute of Urology, The 2 Hospital of Tianjin Medical University, Tianjin Medical University, Tianjin, China
- Department of Psychology, University of Iowa, Iowa City, Iowa, United States of America
| | - Catherine Bradley
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
- Department of Psychology, University of Iowa, Iowa City, Iowa, United States of America
| | - Andrew Schrepf
- Tianjin Institute of Urology, The 2 Hospital of Tianjin Medical University, Tianjin Medical University, Tianjin, China
| | - Liwei Liu
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
- Department of Obstetrics and Gynecology, University of Iowa, Iowa City, Iowa, United States of America
| | - Karl Kreder
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
- Department of Psychology, University of Iowa, Iowa City, Iowa, United States of America
| | - Yi Luo
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
- * E-mail:
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18
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Xu S, Wang X, Wang Y, Lutgendorf S, Bradley C, Schrepf A, Kreder K, O'Donnell M, Luo Y. Transgenic Mice Expressing MCP-1 by the Urothelium Demonstrate Bladder Hypersensitivity, Pelvic Pain and Voiding Dysfunction: A Multidisciplinary Approach to the Study of Chronic Pelvic Pain Research Network Animal Model Study. PLoS One 2016; 11:e0163829. [PMID: 27684718 PMCID: PMC5042429 DOI: 10.1371/journal.pone.0163829] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/10/2016] [Accepted: 09/14/2016] [Indexed: 01/01/2023] Open
Abstract
Monocyte chemoattractant protein-1 (MCP-1) is one of the key chemokines that play important roles in diverse inflammatory and chronic pain conditions. Interstitial cystitis/bladder pain syndrome (IC/BPS) is a chronic and debilitating inflammatory condition of the urinary bladder characterized by the hallmark symptoms of pelvic pain and voiding dysfunction. To facilitate IC/BPS research, we used transgenic technology to develop a novel urothelial MCP-1 secretion mouse model (URO-MCP-1). A transgene consisting of the uroplakin II gene promoter and the mouse MCP-1 coding sequence with a secretory element was constructed and microinjected. URO-MCP-1 mice were found to express MCP-1 mRNA in the bladder epithelium and MCP-1 protein in the urine, and developed bladder inflammation 24 hours after intravesical administration of a single sub-noxious dose of lipopolysaccharide (LPS). The inflamed bladders of URO-MCP-1 mice exhibited elevated mRNAs for interleukin (IL)-1ß, IL-6, substance P precursor, and nerve growth factor as well as increased macrophage infiltration. In parallel with these phenotypic changes, URO-MCP-1 mice manifested significant functional changes at days 1 and 3 after cystitis induction. These functional changes included pelvic pain as measured by von Frey filament stimulation and voiding dysfunction (increased urinary frequency, reduced average volume voided per micturition, and reduced maximum volume voided per micturition) as measured by micturition cages. Micturition changes remained evident at day 7 after cystitis induction, although these changes were not statistically significant. Control wild-type C57BL/6 mice manifested no clear changes in histological, biochemical and behavioral features after similar cystitis induction with LPS. Taken together, our results indicate that URO-MCP-1 mice are hypersensitive to bladder irritants such as LPS and develop pelvic pain and voiding dysfunction upon cystitis induction, providing a novel model for IC/BPS research.
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Affiliation(s)
- Suming Xu
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
| | - Xu Wang
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
| | - Yaoqin Wang
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
| | - Susan Lutgendorf
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
- Department of Psychology, University of Iowa, Iowa City, Iowa, United States of America
- Department of Obstetrics and Gynecology, University of Iowa, Iowa City, Iowa, United States of America
| | - Catherine Bradley
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
- Department of Obstetrics and Gynecology, University of Iowa, Iowa City, Iowa, United States of America
| | - Andrew Schrepf
- Department of Psychology, University of Iowa, Iowa City, Iowa, United States of America
| | - Karl Kreder
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
- Department of Obstetrics and Gynecology, University of Iowa, Iowa City, Iowa, United States of America
| | - Michael O'Donnell
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
| | - Yi Luo
- Department of Urology, University of Iowa, Iowa City, Iowa, United States of America
- * E-mail:
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19
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Liu WJ, Luo Y. Regulatory T cells suppress autoreactive CD4 + T cell response to bladder epithelial antigen. World J Immunol 2016; 6:105-118. [DOI: 10.5411/wji.v6.i2.105] [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: 03/15/2016] [Revised: 04/26/2016] [Accepted: 06/29/2016] [Indexed: 02/05/2023] Open
Abstract
AIM: To investigate the role of regulatory T (Treg) cells in CD4+ T cell-mediated bladder autoimmune inflammation.
METHODS: Urothelium-ovalbumin (URO-OVA)/OT-II mice, a double transgenic line that expresses the membrane form of the model antigen (Ag) OVA as a self-Ag on the urothelium and the OVA-specific CD4+ T cell receptor specific for the I-Ab/OVA323-339 epitope in the periphery, were developed to provide an autoimmune environment for investigation of the role of Treg cells in bladder autoimmune inflammation. To facilitate Treg cell analysis, we further developed URO-OVAGFP-Foxp3/OT-II mice, a derived line of URO-OVA/OT-II mice that express the green fluorescent protein (GFP)-forkhead box protein P3 (Foxp3) fusion protein.
RESULTS: URO-OVA/OT-II mice failed to develop bladder inflammation despite the presence of autoreactive CD4+ T cells. By monitoring GFP-positive cells, bladder infiltration of CD4+ Treg cells was observed in URO-OVAGFP-Foxp3/OT-II mice. The infiltrating Treg cells were functionally active and expressed Treg cell effector molecule as well as marker mRNAs including transforming growth factor-β, interleukin (IL)-10, fibrinogen-like protein 2, and glucocorticoid-induced tumor necrosis factor receptor (GITR). Studies further revealed that Treg cells from URO-OVAGFP-Foxp3/OT-II mice were suppressive and inhibited autoreactive CD4+ T cell proliferation and interferon (IFN)-γ production in response to OVA Ag stimulation. Depletion of GITR-positive cells led to spontaneous development of bladder inflammation and expression of inflammatory factor mRNAs for IFN-γ, IL-6, tumor necrosis factor-α and nerve growth factor in URO-OVAGFP-Foxp3/OT-II mice.
CONCLUSION: Treg cells specific for bladder epithelial Ag play an important role in immunological homeostasis and the control of CD4+ T cell-mediated bladder autoimmune inflammation.
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20
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Aich A, Afrin LB, Gupta K. Mast Cell-Mediated Mechanisms of Nociception. Int J Mol Sci 2015; 16:29069-92. [PMID: 26690128 PMCID: PMC4691098 DOI: 10.3390/ijms161226151] [Citation(s) in RCA: 100] [Impact Index Per Article: 10.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/04/2015] [Revised: 11/28/2015] [Accepted: 12/01/2015] [Indexed: 12/12/2022] Open
Abstract
Mast cells are tissue-resident immune cells that release immuno-modulators, chemo-attractants, vasoactive compounds, neuropeptides and growth factors in response to allergens and pathogens constituting a first line of host defense. The neuroimmune interface of immune cells modulating synaptic responses has been of increasing interest, and mast cells have been proposed as key players in orchestrating inflammation-associated pain pathobiology due to their proximity to both vasculature and nerve fibers. Molecular underpinnings of mast cell-mediated pain can be disease-specific. Understanding such mechanisms is critical for developing disease-specific targeted therapeutics to improve analgesic outcomes. We review molecular mechanisms that may contribute to nociception in a disease-specific manner.
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Affiliation(s)
- Anupam Aich
- Vascular Biology Center, Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA.
| | - Lawrence B Afrin
- Vascular Biology Center, Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA.
| | - Kalpna Gupta
- Vascular Biology Center, Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA.
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21
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Lai H, Gereau RW, Luo Y, O'Donnell M, Rudick CN, Pontari M, Mullins C, Klumpp DJ. Animal Models of Urologic Chronic Pelvic Pain Syndromes: Findings From the Multidisciplinary Approach to the Study of Chronic Pelvic Pain Research Network. Urology 2015; 85:1454-65. [PMID: 26099889 DOI: 10.1016/j.urology.2015.03.007] [Citation(s) in RCA: 36] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/14/2014] [Revised: 02/13/2015] [Accepted: 03/06/2015] [Indexed: 11/16/2022]
Abstract
OBJECTIVE To describe the approach taken by the Multidisciplinary Approach to the Study of Chronic Pelvic Pain Research Network investigators to advance the utility of urologic chronic pelvic pain syndromes (UCPPS) animal models. METHODS A multidisciplinary team of investigators representing basic science and clinical expertise defined key phenotypic criteria for rodent models of UCPPS. UCPPS symptoms were prioritized based on their clinical significance. Methods for quantifying animal correlates to patient symptoms were developed. The methods were implemented across proposed rodent models for evaluation and comparison of animals for phenotypic characteristics relevant to human symptomatology. RESULTS Pelvic pain and urinary frequency were deemed primary features of human UCPPS and were prioritized for assessment in animals. Nociception was quantified using visceromotor response to bladder distention and by applying von Frey filaments to the lower abdomen (referred tactile allodynia). Micturition activity was assessed as free voiding using micturition cages or blotting pad assays and in response to bladder filling by cystometry. Models varied in both depth of characterization and degree of recapitulating pelvic pain and urinary frequency characteristics of UCPPS. CONCLUSION Rodent models that reflect multiple key characteristics of human UCPPS may be identified and provide enhanced clinical significance to mechanistic studies. We have developed a strategy for evaluating current and future animal models of UCPPS based on human symptomatology. This approach provides a foundation for improved translation between mechanistic studies in animals and clinical research and serves as a validation strategy for assessing validity of models for symptom-driven disorders of unknown etiology.
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Affiliation(s)
- Henry Lai
- Division of Urologic Surgery, Department of Surgery, Washington University School of Medicine, St Louis, MO.
| | - Robert W Gereau
- Department of Anesthesiology, Washington University School of Medicine, St Louis, MO
| | - Yi Luo
- Department of Urology, University of Iowa Carver College of Medicine, Iowa City, IA
| | - Michael O'Donnell
- Department of Urology, University of Iowa Carver College of Medicine, Iowa City, IA
| | - Charles N Rudick
- Department of Urology, Northwestern University Feinberg School of Medicine, Chicago, IL
| | - Michel Pontari
- Department of Urology, Temple University School of Medicine, Philadelphia, PA
| | - Chris Mullins
- Division of Kidney, Urologic, & Hematologic Diseases (KUH), National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health, Bethesda, MD
| | - David J Klumpp
- Department of Urology, Northwestern University Feinberg School of Medicine, Chicago, IL
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Izgi K, Altuntas CZ, Bicer F, Ozer A, Sakalar C, Li X, Tuohy VK, Daneshgari F. Uroplakin peptide-specific autoimmunity initiates interstitial cystitis/painful bladder syndrome in mice. PLoS One 2013; 8:e72067. [PMID: 23977210 PMCID: PMC3745386 DOI: 10.1371/journal.pone.0072067] [Citation(s) in RCA: 28] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/01/2013] [Accepted: 07/09/2013] [Indexed: 12/27/2022] Open
Abstract
The pathophysiology of interstitial cystitis/painful bladder syndrome (IC/PBS) is enigmatic. Autoimmunity and impaired urothelium might lead the underlying pathology. A major shortcoming in IC/PBS research has been the lack of an appropriate animal model. In this study, we show that the bladder specific uroplakin 3A-derived immunogenic peptide UPK3A 65–84, which contains the binding motif for IAd MHC class II molecules expressed in BALB/c mice, is capable of inducing experimental autoimmune cystitis in female mice of that strain. A highly antigen-specific recall proliferative response of lymph node cells to UPK3A 65–84 was observed, characterized by selectively activated CD4+ T cells with a proinflammatory Th1-like phenotype, including enhanced production of interferon γ and interleukin-2. T cell infiltration of the bladder and bladder-specific increased gene expression of inflammatory cytokines were observed. Either active immunization with UPK3A 65–84 or adoptive transfer of peptide-activated CD4+ T cells induced all of the predominant IC/PBS phenotypic characteristics, including increased micturition frequency, decreased urine output per micturition, and increased pelvic pain responses to stimulation with von Frey filaments. Our study demonstrates the creation of a more specific experimental autoimmune cystitis model that is the first inducible model for IC/PBS that manifests all of the major symptoms of this debilitating condition.
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Affiliation(s)
- Kenan Izgi
- Department of Urology, Case Western Reserve University, Cleveland, Ohio, United States of America
- Department of Clinical Chemistry, Cleveland State University, Cleveland,, Ohio, United States of America
| | - Cengiz Z. Altuntas
- Department of Urology, Case Western Reserve University, Cleveland, Ohio, United States of America
| | - Fuat Bicer
- Department of Urology, Case Western Reserve University, Cleveland, Ohio, United States of America
- Department of Clinical Chemistry, Cleveland State University, Cleveland,, Ohio, United States of America
| | - Ahmet Ozer
- Department of Urology, Case Western Reserve University, Cleveland, Ohio, United States of America
- Department of Genetics, Case Western Reserve University, Cleveland, Ohio, United States of America
| | - Cagri Sakalar
- Department of Immunology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, United States of America
| | - Xiaoxia Li
- Department of Immunology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, United States of America
| | - Vincent K. Tuohy
- Department of Immunology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, United States of America
| | - Firouz Daneshgari
- Department of Urology, Case Western Reserve University, Cleveland, Ohio, United States of America
- * E-mail:
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BALB/c-Fcgr2bPdcd1 mouse expressing anti-urothelial antibody is a novel model of autoimmune cystitis. Sci Rep 2012; 2:317. [PMID: 22432050 PMCID: PMC3307042 DOI: 10.1038/srep00317] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/31/2011] [Accepted: 02/23/2012] [Indexed: 01/22/2023] Open
Abstract
We report the impact of anti-urothelial autoantibody (AUAb) on urinary bladder phenotype in BALB/c mice deficient of the FcγRIIb and PD-1. AUAb was present in serum samples from approximately half of the double-knockout (DKO) mice, as detected by immunofluorescence and immunoblots for urothelial proteins including uroplakin IIIa. The AUAb-positive DKO mice showed degeneration of urothelial plaque and umbrella cells, along with infiltration of inflammatory cells in the suburothelial layer. TNFα and IL-1β were upregulated in the bladder and the urine of AUAb-positive DKO mice. Voiding behavior of mice was analyzed by the Voided Stain on Paper method. 10-week-old and older AUAb-positive DKO mice voided significantly less urine per void than did wild type (WT) mice. Furthermore, administration of the AUAb-containing serum to WT mice significantly reduced their urine volume per void. In summary, this report presents a novel comprehensive mouse model of autoimmune cystitis.
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24
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Reduction of intercellular adhesion molecule 1 may play a role in anti-inflammatory effect of hyaluronic acid in a rat model of severe non-bacterial cystitis. World J Urol 2012; 31:535-40. [PMID: 22358112 DOI: 10.1007/s00345-012-0839-8] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/12/2011] [Accepted: 02/07/2012] [Indexed: 10/28/2022] Open
Abstract
PURPOSE To evaluate the impact of intercellular adhesion molecule 1 (ICAM-1) in hyaluronic acid (HA) therapy in rats model of severe non-bacterial cystitis. METHODS Cystitis models in Sprague-Dawley female rats were produced by combination of intraperitoneal cyclophosphamide (CYP) with intravesical protamine/lipopolysaccharide (PS/LPS). HA or heparin (0.5 ml) was introduced intravesically to rats' bladders followed PS/LPS. Bladder tissue was prepared for histology including mast cell presence and measurement of ICAM-1, tumor necrosis factor (TNF)-α, and interleukin 6 (IL-6). RESULTS Cystitis model using intraperitoneal CYP and intravesical SP/LPS showed serious inflammation, higher mast cell count with elevated ICAM-1, TNF-α, and IL-6 levels. After intravesical heparin or HA treatment, incidence of grades 3-4 bladder inflammation and tissue ICAM-1 level were only significantly lower in HA group (P = 0.017, P = 0.021, respectively), but not in heparin group (P = 0.12, P = 0.798, respectively). Remarkably lower level of TNF-α (P = 0.003) and ICAM-1 (P = 0.006) was detected in HA-treated rats compared with heparin-treated rats. Inflammation grade and ICAM-1 level had strong correlation (P < 0.001). IL-6 level after HA or heparin instillation had no difference. CONCLUSIONS Intravesical administration of HA decreased the severity of bladder inflammation, mast cell presence, and levels of ICAM-1 and TNF-α in a rat model of severe non-bacterial cystitis; its effect was more obvious than that of heparin. Reduction of ICAM-1 may play a role in the anti-inflammatory effect of HA.
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25
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Urothelial antigen-specific CD4+ T cells function as direct effector cells and induce bladder autoimmune inflammation independent of CD8+ T cells. Mucosal Immunol 2011; 4:428-37. [PMID: 21270773 PMCID: PMC3118865 DOI: 10.1038/mi.2010.90] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023]
Abstract
The role of CD4(+) T cells in bladder autoimmune inflammation has not been identified because of the lack of a proper animal model. We investigated CD4(+) T-cell responses to bladder urothelial ovalbumin (OVA), a model self-antigen (Ag), in transgenic URO-OVA mice. The expression of bladder urothelial OVA rendered mice unresponsive to OVA and resulted in quick clearance of Ag-specific CD4(+) T cells. Adoptive transfer of naive OVA-specific CD4(+) T cells led to exogenous T-cell proliferation, activation, and bladder infiltration but no inflammatory induction. In contrast, adoptive transfer of preactivated OVA-specific CD4(+) T cells induced bladder inflammation. Studies further demonstrated that CD4(+) T cells induced bladder inflammation in URO-OVA mice depleted of CD8(+) T cells or deficient in the recombinase activating gene-1 (Rag-1(-/-)). These results indicate that urothelial Ag-specific CD4(+) T cells can function as direct effector cells to induce bladder autoimmune inflammation independent of CD8(+) T cells.
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26
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Abstract
Inflammation of the lower urinary tract occurs frequently in people. The causes remain obscure, with the exception of urinary tract infection. Animal models have proven useful for investigating and assessing mechanisms underlying symptoms associated with lower urinary tract inflammation and options for suppressing these symptoms. This review will discuss various animal models of lower urinary tract inflammation, including feline spontaneous (interstitial) cystitis, neurogenic cystitis, autoimmune cystitis, cystitis induced by intravesical instillation of chemicals or bacterial products (particularly lipopolysaccharide or LPS), and prostatic inflammation initiated by transurethral instillation of bacteria. Animal models will continue to be of significant value in identifying mechanisms resulting in bladder inflammation, but the relevance of some of these models to the causes underlying clinical disease is unclear. This is primarily because of the lack of understanding of causes of these disorders in people. Comparative and translational studies are required if the full potential of findings obtained with animal models to improve prevention and treatment of lower urinary tract inflammation in people is to be realized.
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Affiliation(s)
- Dale E Bjorling
- Department of Surgical Sciences, University of Wisconsin-Madison, Madison, WI, USA.
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27
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Intravesical dimethyl sulfoxide inhibits acute and chronic bladder inflammation in transgenic experimental autoimmune cystitis models. J Biomed Biotechnol 2010; 2011:937061. [PMID: 21113298 PMCID: PMC2989383 DOI: 10.1155/2011/937061] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/08/2010] [Accepted: 10/20/2010] [Indexed: 02/08/2023] Open
Abstract
New animal models are greatly needed in interstitial cystitis/painful bladder syndrome (IC/PBS) research. We recently developed a novel transgenic cystitis model (URO-OVA mice) that mimics certain key aspects of IC/PBS pathophysiology. This paper aimed to determine whether URO-OVA cystitis model was responsive to intravesical dimethyl sulfoxide (DMSO) and if so identify the mechanisms of DMSO action. URO-OVA mice developed acute cystitis upon adoptive transfer of OVA-specific OT-I splenocytes. Compared to PBS-treated bladders, the bladders treated with 50% DMSO exhibited markedly reduced bladder histopathology and expression of various inflammatory factor mRNAs. Intravesical DMSO treatment also effectively inhibited bladder inflammation in a spontaneous chronic cystitis model (URO-OVA/OT-I mice). Studies further revealed that DMSO could impair effector T cells in a dose-dependent manner in vitro. Taken together, our results suggest that intravesical DMSO improves the bladder histopathology of IC/PBS patients because of its ability to interfere with multiple inflammatory and bladder cell types.
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28
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Wang ZY, Wang P, Bjorling DE. Role of mast cells and protease-activated receptor-2 in cyclooxygenase-2 expression in urothelial cells. Am J Physiol Regul Integr Comp Physiol 2009; 297:R1127-35. [PMID: 19675284 DOI: 10.1152/ajpregu.00310.2009] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/30/2022]
Abstract
Mast cells have been shown to play a role in development and persistence of various inflammatory bladder disorders. Mast cell-derived tryptase specifically activates protease-activated receptor-2 (PAR-2), and PAR-2 is known to be involved in inflammation. We investigated whether mast cells participate in increase of cyclooxygenase-2 (COX-2) protein abundance in urothelium/suburothelium of bladders of mice subsequent to cyclophosphamide (CYP)-induced bladder inflammation. We also used primary cultures of human urothelial cells to investigate cellular mechanisms underlying activation of PAR-2 resulting in increased COX-2 expression. We found that treatment of mice with CYP (150 mg/kg ip) increased COX-2 protein abundance in bladder urothelium/suburothelium 3, 6, and 24 h after CYP (P < 0.01), and increased COX-2 protein abundance was prevented by treatment of mice with the mast cell stabilizer sodium cromolyn (10 mg/kg ip) for 4 consecutive days before CYP treatment. Incubation of freshly isolated mouse urothelium/suburothelium with a selective PAR-2 agonist, 2-furoyl-LIGRLO-amide (3 microM), also increased COX-2 protein abundance (P < 0.05). We further demonstrated that 2-furoyl-LIGRLO-amide (3 microM) increased COX-2 mRNA expression and protein abundance in primary cultures of human urothelial cells (P < 0.01), and the effects of PAR-2 activation were mediated primarily by the ERK1/2 MAP kinase pathway. These data indicate that there are functional interactions among mast cells, PAR-2 activation, and increased expression of COX-2 in bladder inflammation.
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Affiliation(s)
- Zun-Yi Wang
- Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin 53719, USA.
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29
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Liu W, Deyoung BR, Chen X, Evanoff DP, Luo Y. RDP58 inhibits T cell-mediated bladder inflammation in an autoimmune cystitis model. J Autoimmun 2008; 30:257-65. [PMID: 18162370 PMCID: PMC2441447 DOI: 10.1016/j.jaut.2007.10.005] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/17/2007] [Revised: 10/20/2007] [Accepted: 10/22/2007] [Indexed: 12/28/2022]
Abstract
Interstitial cystitis (IC) is a chronic inflammatory condition of the urinary bladder with a strong autoimmune component. Currently, the major challenge in IC treatment is the development of effective therapies. RDP58 is a novel d-amino acid decapeptide with potent immunosuppressive activity. In this study, we investigated whether RDP58 was effective as an intravesical agent for treating bladder autoimmune inflammation in a transgenic mouse model (URO-OVA mice). URO-OVA mice were adoptively transferred with syngeneic activated splenocytes of OT-I mice transgenic for the OVA-specific CD8(+) TCR for cystitis induction and treated intravesically with RDP58 at days 0 and 3. Compared with controls, the RDP58-treated bladders showed markedly reduced histopathology and expressions of mRNAs and proteins of TNF-alpha, NGF and substance P. To determine whether the inhibition of bladder inflammation by RDP58 was due to the interference with effector T cells, we treated the cells with RDP58 in vitro. Cells treated with RDP58 showed reduced production of TNF-alpha and IFN-gamma as well as apoptotic death. Collectively, these results indicate that RDP58 is effective for treating T cell-mediated experimental autoimmune cystitis and may serve as a useful intravesical agent for the treatment of autoimmune-associated bladder inflammation such as IC.
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Affiliation(s)
- Wujiang Liu
- Department of Urology, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, 3202 MERF, 375 Newton Road, IA 52242-1087, USA
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