Federal State Budgetary Scientific Institution "Research Institute of Fundamental and Clinical Immunology" (RIFCI), 630099 Novosibirsk, Russia.
State Research Center of Virology and Biotechnology "Vector", 630559 Koltsovo, Russia.
Front Biosci (Landmark Ed). 2022 May 30;27(6):170. doi: 10.31083/j.fbl2706170.
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 , using dendritic cells (DCs) transfected with designed DNA constructs encoding the stimulator's major histocompatibility complex (MHC) epitopes.
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).
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.
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.
寻找有效的移植物排斥调节技术以促进持久的移植物植入仍然是全世界密切研究的课题。尽管有各种免疫抑制剂，但目前使用的方案缺乏选择性，并且有许多副作用。在这里，我们研究了一种在人类“刺激物-应答者”模型中诱导抗原特异性耐受的方法，使用转染了设计的 DNA 构建体的树突状细胞（DC），该构建体编码刺激物的主要组织相容性复合物（MHC）表位。
本研究的对象是来自 10 名健康供体的外周血单核细胞（PBMC）。为了诱导抗原特异性耐受，根据供体和受者 MHC 的序列，为五个应答器-刺激器对创建了个性化的 DNA 构建体。进行 DNA 测序以选择要纳入遗传构建体的表位。使用混合淋巴细胞培养测定法（i）评估所有可能的刺激物-应答器对（90 个反应）的双向增殖反应，以及（ii）评估生成的转染 DC 的耐受性（5 个反应）。
与用供体特异性质粒转染的应答者树突状细胞共培养后，供体单核细胞培养物中的 FoxP3+CD4+CD25+细胞数量和 IL-10 产生明显增加。使用转染 MHC 表位的耐受性 DC 生成的耐受性培养物具有更大的能力抑制自体 MNC 对同种异体 MHC 刺激的增殖。
转染针对 HLA 刺激表位的 DNA 构建体的产生的 DC 表现出耐受性，并可用于开发抗原特异性耐受。因此，我们提出了一种诱导抗原特异性耐受的有前途的方法，随后应在临床实践中进行研究。
