TY - JOUR

T1 - In Vitro Model of Suppression of the Alloantigen Response by Tolerogenic Dendritic Cells Transfected with Personalized DNA Constructs Encoding HLA Epitopes

AU - Shevchenko, Julia A.

AU - Lopatnikova, Julia A.

AU - Khantakova, Julia N.

AU - Silkov, Alexander N.

AU - Kuznetsova, Maria S.

AU - Kurilin, Vasiliy V.

AU - Maksyutov, Amir Z.

AU - Sennikov, Sergey V.

N1 - Funding Information: The research (MLC, culture work, functional assays) was conducted under the framework of State Assignment № 1021062512015-4 (FGMN-2021-0003). The steps of DNA transduction and HLA-A genotyping were sponsored by the grant of the Russian Science Foundation project no. 21-65-00004, https://rscf.ru/project/21-65-00004/. Publisher Copyright: Copyright: © 2022 The Author(s)

PY - 2022/6

Y1 - 2022/6

N2 - Background: A search for efficient graft rejection modulation techniques for the promotion of durable engraftment remains to be a matter of close study all over the world. Despite the variety of immunosuppressive drugs, the schemes currently used show a lack of selectivity and have a number of side effects. Here we investigated an approach for the induction of antigen-specific tolerance in a human “stimulator-responder” model in vitro, using dendritic cells (DCs) transfected with designed DNA constructs encoding the stimulator's major histocompatibility complex (MHC) epitopes. Methods: The object of the study is peripheral blood mononuclear cells (PBMCs) from 10 healthy donors. To induce antigen-specific tolerance, personalized DNA constructs were created for five responder-stimulator pairs, based on the sequences of donors' and recipients' MHCs. DNA sequencing was performed to select epitopes for incorporation into genetic constructs. A mixed lymphocyte culture assay was used (i) to assess the proliferative response in both directions for all possible stimulator-responder pairs (90 reactions) and (ii) to assess the tolerogenic properties of the generated transfected DCs (5 reactions). Results: A significant increase in the amounts of FoxP3+ CD4+CD25+ cells and in IL-10 production was shown in culture of donor mononuclear cells after co-cultivation with the responder's dendritic cells transfected with donor-specific plasmids. The tolerogenic cultures generated using tolerogenic DCs transfected with MHC epitopes had a significantly greater ability to inhibit the proliferation of autologous MNCs in response to an allogeneic MHC stimulus. Conclusions: The produced DCs transfected with DNA constructs against HLA stimulating epitopes exhibited tolerogenic properties and may be used to develop antigen-specific tolerance. Thus, we proposed a perspective approach to the induction of antigen-specific tolerance, which should subsequently be studied for use in clinical practice.

AB - Background: A search for efficient graft rejection modulation techniques for the promotion of durable engraftment remains to be a matter of close study all over the world. Despite the variety of immunosuppressive drugs, the schemes currently used show a lack of selectivity and have a number of side effects. Here we investigated an approach for the induction of antigen-specific tolerance in a human “stimulator-responder” model in vitro, using dendritic cells (DCs) transfected with designed DNA constructs encoding the stimulator's major histocompatibility complex (MHC) epitopes. Methods: The object of the study is peripheral blood mononuclear cells (PBMCs) from 10 healthy donors. To induce antigen-specific tolerance, personalized DNA constructs were created for five responder-stimulator pairs, based on the sequences of donors' and recipients' MHCs. DNA sequencing was performed to select epitopes for incorporation into genetic constructs. A mixed lymphocyte culture assay was used (i) to assess the proliferative response in both directions for all possible stimulator-responder pairs (90 reactions) and (ii) to assess the tolerogenic properties of the generated transfected DCs (5 reactions). Results: A significant increase in the amounts of FoxP3+ CD4+CD25+ cells and in IL-10 production was shown in culture of donor mononuclear cells after co-cultivation with the responder's dendritic cells transfected with donor-specific plasmids. The tolerogenic cultures generated using tolerogenic DCs transfected with MHC epitopes had a significantly greater ability to inhibit the proliferation of autologous MNCs in response to an allogeneic MHC stimulus. Conclusions: The produced DCs transfected with DNA constructs against HLA stimulating epitopes exhibited tolerogenic properties and may be used to develop antigen-specific tolerance. Thus, we proposed a perspective approach to the induction of antigen-specific tolerance, which should subsequently be studied for use in clinical practice.

KW - alloantigens

KW - DNA constructs

KW - HLA

KW - mediated immune suppression

KW - MHC

KW - mixed lymphocyte culture

KW - T regulatory cells

KW - tolerogenic dendritic cells

KW - Humans

KW - Dendritic Cells/metabolism

KW - Leukocytes, Mononuclear

KW - Epitopes/genetics

KW - Isoantigens/genetics

KW - Immune Tolerance/genetics

KW - T-Lymphocytes, Regulatory

UR - http://www.scopus.com/inward/record.url?scp=85132936182&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/08acfa01-705a-3cbe-9e21-9e6e35990d50/

U2 - 10.31083/j.fbl2706170

DO - 10.31083/j.fbl2706170

M3 - Article

C2 - 35748246

AN - SCOPUS:85132936182

VL - 27

JO - Frontiers in Bioscience - Landmark

JF - Frontiers in Bioscience - Landmark

SN - 2768-6701

IS - 6

M1 - 170

ER -