Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Nov 26, 2020; 12(11): 1377-1395
Published online Nov 26, 2020. doi: 10.4252/wjsc.v12.i11.1377
Immunophenotypic characteristics of multipotent mesenchymal stromal cells that affect the efficacy of their use in the prevention of acute graft vs host disease
Nataliya Petinati, Nikolay Kapranov, Yulia Davydova, Alexey Bigildeev, Olesya Pshenichnikova, Dmitriy Karpenko, Nina Drize, Larisa Kuzmina, Elena Parovichnikova, Valeriy Savchenko
Nataliya Petinati, Alexey Bigildeev, Dmitriy Karpenko, Nina Drize, Laboratory for Physiology of Hematopoiesis, National Research Center for Hematology, Moscow 125167, Russia
Nikolay Kapranov, Yulia Davydova, Laboratory for Immunophenotyping of Blood and Bone Marrow Cells, National Research Center for Hematology, Moscow 125167, Russia
Olesya Pshenichnikova, Laboratory for Genetic Engineering, National Research Center for Hematology, Moscow 125167, Russia
Larisa Kuzmina, Elena Parovichnikova, Valeriy Savchenko, Hematopoiesis Depression and Bone Marrow Transplantation Department, National Research Center for Hematology, Moscow 125167, Russia
Author contributions: Petinati N, Drize N and Savchenko V designed and coordinated the study; Petinati N, Kapranov N, Davydova Y, Bigildeev A and Drize N performed the experiments, acquired and analyzed data; Kuzmina L and Parovichnikova E collected clinical data; Pshenichnikova O and Karpenko D performed the statistical analysis; Petinati N and Drize N wrote the manuscript; all authors approved the final version of the article.
Supported by Russian Foundation for Basic Research, No. 19-29-04023.
Institutional review board statement: The study was reviewed and approved by the National Research Center for Hematology, Moscow, Russia Institutional Review Board (Approval No. 140, 18 April 2019).
Conflict-of-interest statement: The authors declare that they have no conflict of interest.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Nina Drize, PhD, Professor, Laboratory for Physiology of Hematopoiesis, National Research Center for Hematology, Noviy Zykovskiy, 4, Moscow 125167, Russia. drize.n@blood.ru
Received: May 28, 2020
Peer-review started: May 28, 2020
First decision: July 30, 2020
Revised: July 31, 2020
Accepted: September 1, 2020
Article in press: September 1, 2020
Published online: November 26, 2020
Processing time: 178 Days and 1.9 Hours
Abstract
BACKGROUND

Multipotent mesenchymal stromal cells (MSCs) are widely used in the clinic due to their unique properties, namely, their ability to differentiate in all mesenchymal directions and their immunomodulatory activity. Healthy donor MSCs were used to prevent the development of acute graft vs host disease (GVHD) after allogeneic bone marrow transplantation (allo-BMT). The administration of MSCs to patients was not always effective. The MSCs obtained from different donors have individual characteristics. The differences between MSC samples may affect their clinical efficacy.

AIM

To study the differences between effective and ineffective MSCs.

METHODS

MSCs derived from the bone marrow of a hematopoietic stem cells donor were injected intravenously into allo-BMT recipients for GVHD prophylaxis at the moment of blood cell reconstitution. Aliquots of 52 MSC samples that were administered to patients were examined, and the same cells were cultured in the presence of peripheral blood mononuclear cells (PBMCs) from a third-party donor or treated with the pro-inflammatory cytokines IL-1β, IFN and TNF. Flow cytometry revealed the immunophenotype of the nontreated MSCs, the MSCs cocultured with PBMCs for 4 d and the MSCs exposed to cytokines. The proportions of CD25-, CD146-, CD69-, HLA-DR- and PD-1-positive CD4+ and CD8+ cells and the distribution of various effector and memory cell subpopulations in the PBMCs cocultured with the MSCs were also determined.

RESULTS

Differences in the immunophenotypes of effective and ineffective MSCs were observed. In the effective samples, the mean fluorescence intensity (MFI) of HLA-ABC, HLA-DR, CD105, and CD146 was significantly higher. After MSCs were treated with IFN or cocultured with PBMCs, the HLA-ABC, HLA-DR, CD90 and CD54 MFI showed a stronger increase in the effective MSCs, which indicated an increase in the immunomodulatory activity of these cells. When PBMCs were cocultured with effective MSCs, the proportions of CD4+ and CD8+central memory cells significantly decreased, and the proportion of CD8+CD146+ lymphocytes increased more than in the subpopulations of lymphocytes cocultured with MSC samples that were ineffective in the prevention of GVHD; in addition, the proportion of CD8+effector memory lymphocytes decreased in the PBMCs cocultured with the effective MSC samples but increased in the PBMCs cocultured with the ineffective MSC samples. The proportion of CD4+CD146+ lymphocytes increased only when cocultured with the inefficient samples.

CONCLUSION

For the first time, differences were observed between MSC samples that were effective for GVHD prophylaxis and those that were ineffective. Thus, it was shown that the immunomodulatory activity of MSCs depends on the individual characteristics of the MSC population.

Keywords: Multipotent mesenchymal stromal cells; Acute graft vs host disease; Immunophenotype; Lymphocytes; Immunomodulation; Pro-inflammatory cytokines

Core Tip: An attempt was made to identify the main differences between multipotent mesenchymal stromal cells (MSCs) samples that are effective and those that are ineffective in preventing the development of acute graft vs host disease after allogeneic bone marrow transplantation. The mean fluorescence intensity of HLA-ABC, HLA-DR, CD105, and CD146 was shown to be significantly lower on the surface of samples that were ineffective for prophylaxis. Significant differences were revealed between effective and ineffective MSCs in terms of their responses to interaction with lymphocytes and stimulation by pro-inflammatory cytokines. The patterns observed here indicate a possible mechanism of the immunosuppressive action of these cells in clinical use.