Production of Cytotoxic Antibodies After Intra-Articular Injection of Allogeneic Synovial Membrane Mesenchymal Stem Cells With and Without LPS Administration.
Abstract: Allogeneic mesenchymal stem cells (MSC) are widely used in clinical routine due to the shorter expansion time and reliability of its quality. However, some recipients can produce alloantibodies that recognize MSCs and activate the immune system, resulting in cell death. Although antibody production was already described after MSC injection, no previous studies described the immune response after intra-articular MSC injection in acute synovitis. This study aimed to evaluate the influence of inflammation on immune response after single and repeated intra-articular injections of synovial membrane MSC (SMMSC). Horses were divided in three groups: control group (AUTO) received autologous synovial membrane MSCs; whereas group two (ALLO) received allogeneic SMMSCs and group three (ALLO LPS) was submitted to acute experimental synovitis 8 h before SMMSCs injection. The procedure was repeated for all groups for 28 days. Physical and lameness evaluations and synovial fluid analysis were performed. Sera from all animals were obtained before and every 7 days after each injection up to 4 weeks, to perform microcytotoxicity assays incubating donor SMMSCs with recipients' sera. The first injection caused a mild and transient synovitis in all groups, becoming more evident and longer in ALLO and ALLO LPS groups after the second injection. Microcytotoxicity assays revealed significant antibody production as soon as 7 days after SMMSC injection in ALLO and ALLO LPS groups, and cytotoxicity scores of both groups showed no differences at any time point, being equally different from AUTO group. Although inflammation is capable of inducing MHC expression in MSCs, which enhances immune recognition, cytotoxicity scores were equally high in ALLO and ALLO LPS groups, making it difficult to determine the potentiation effect of inflammation on antibody production. Our findings suggest that inflammation does not display a pivotal role in immune recognition on first allogeneic MSC injection. In a translational way, since specific antibodies were produced against MSCs, patients that need more than one MSC injection may benefit from a first allogeneic injection followed by subsequent autologous injections.
Copyright © 2022 Rosa, Krieck, Padula, Stievani, Rossi, Pfeifer, Basso, Braz, Golim and Alves.
Publication Date: 2022-04-27 PubMed ID: 35572507PubMed Central: PMC9091817DOI: 10.3389/fimmu.2022.871216Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates the antibody response to single and repeated injections of allogeneic synovial membrane mesenchymal stem cells (SMMSCs) in the treatment of acute synovitis. It suggests that inflammation does not play a major role in the immune response against the first allogeneic MSC injection, and patients requiring multiple MSC injections might benefit from an initial allogeneic injection succeeded by autologous injections.
Objective and Methodology
- The study aims to understand how inflammation impacts the immune response after single or repeated injections of SMMSCs directly into the joint (intra-articular).
- The experiment consisted of three groups of horses: a control group (AUTO) received autologous SMMSCs, ALLO received allogeneic SMMSCs, and ALLO LPS underwent acute experimental synovitis before receiving the SMMSCs injection.
- This procedure was replicated for all groups over 28 days, with physical and lameness evaluations and synovial fluid analysis conducted.
- Blood samples were collected from all participants before and after each injection at 7-day intervals for analysis of microcytotoxicity to determine antibody production.
Key Findings
- All groups demonstrated a slight and brief inflammation after the first injection, with the ALLO and ALLO LPS groups showing more pronounced and extended inflammation after the second injection.
- Evidence of significant antibody production was noticed as early as 7 days post injection in both the ALLO and ALLO LPS groups, with no remarkable variations in cytotoxicity scores among them.
- Despite the potential of inflammation to enhance immune recognition via MHC expression in MSCs, the cytotoxicity levels were nearly identical in the ALLO and ALLO LPS groups, obscuring any potential effect of inflammation on antibody production.
Implications
- The study suggests that inflammation does not play a significant role in immune recognition during the first allogeneic MSC injection.
- In terms of medical application, as specific antibodies were produced against MSCs, patients requiring multiple MSC injections might benefit from an initial allogeneic injection, followed by autologous injections.
Cite This Article
APA
Rosa GDS, Krieck AMT, Padula ET, Stievani FC, Rossi MC, Pfeifer JPH, Basso RM, Braz AMM, Golim MA, Alves ALG.
(2022).
Production of Cytotoxic Antibodies After Intra-Articular Injection of Allogeneic Synovial Membrane Mesenchymal Stem Cells With and Without LPS Administration.
Front Immunol, 13, 871216.
https://doi.org/10.3389/fimmu.2022.871216 Publication
Researcher Affiliations
- Department of Veterinary Surgery and Animal Reproduction, Regenerative Medicine Lab, School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Botucatu, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, Regenerative Medicine Lab, School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Botucatu, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, Regenerative Medicine Lab, School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Botucatu, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, Regenerative Medicine Lab, School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Botucatu, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, Regenerative Medicine Lab, School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Botucatu, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, Regenerative Medicine Lab, School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Botucatu, Brazil.
- Department of Veterinary Clinics, School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Botucatu, Brazil.
- Flow Cytometry Laboratory, Applied Biotechnology Laboratory, Clinical Hospital of Botucatu Medical School, Botucatu, Brazil.
- Flow Cytometry Laboratory, Applied Biotechnology Laboratory, Clinical Hospital of Botucatu Medical School, Botucatu, Brazil.
- Graduate Program in Research and Development (Medical Biotechnology), Botucatu Medical School, São Paulo State University (UNESP), Botucatu, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, Regenerative Medicine Lab, School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Botucatu, Brazil.
MeSH Terms
- Animals
- Hematopoietic Stem Cell Transplantation / adverse effects
- Horses
- Humans
- Inflammation / complications
- Injections, Intra-Articular / adverse effects
- Lipopolysaccharides
- Mesenchymal Stem Cell Transplantation / methods
- Mesenchymal Stem Cells
- Reproducibility of Results
- Synovial Membrane
- Synovitis / chemically induced
- Synovitis / therapy
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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