Peters Jorieke H, Hilbrands Luuk B, Koenen Hans J P M, Joosten Irma
Department of Bloodtransfusion and Transplantation Immunology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
PLoS One. 2008 May 21;3(5):e2233. doi: 10.1371/journal.pone.0002233.
Regulatory T cell (Treg) based immunotherapy is a potential treatment for several immune disorders. By now, this approach proved successful in preclinical animal transplantation and auto-immunity models. In these models the success of Treg based immunotherapy crucially depends on the antigen-specificity of the infused Treg population. For the human setting, information is lacking on how to generate Treg with direct antigen-specificity ex vivo to be used for immunotherapy.
METHODOLOGY/PRINCIPAL FINDINGS: Here, we demonstrate that in as little as two stimulation cycles with HLA mismatched allogeneic stimulator cells and T cell growth factors a very high degree of alloantigen-specificity was reached in magnetic bead isolated human CD4(pos)CD25(high) Treg. Efficient increases in cell numbers were obtained. Primary allogeneic stimulation appeared a prerequisite in the generation of alloantigen-specific Treg, while secondary allogeneic or polyclonal stimulation with anti-CD3 plus anti-CD28 monoclonal antibodies enriched alloantigen-specificity and cell yield to a similar extent.
CONCLUSIONS/SIGNIFICANCE: The ex vivo expansion protocol that we describe will very likely increase the success of clinical Treg-based immunotherapy, and will help to induce tolerance to selected antigens, while minimizing general immune suppression. This approach is of particular interest for recipients of HLA mismatched transplants.
基于调节性T细胞（Treg）的免疫疗法是治疗多种免疫紊乱疾病的一种潜在方法。到目前为止，这种方法在临床前动物移植和自身免疫模型中已被证明是成功的。在这些模型中，基于Treg的免疫疗法的成功关键取决于注入的Treg群体的抗原特异性。对于人体情况，目前尚缺乏关于如何在体外产生具有直接抗原特异性的Treg用于免疫治疗的信息。
方法/主要发现：在此，我们证明，在与HLA不匹配的同种异体刺激细胞和T细胞生长因子进行短短两个刺激周期后，磁珠分离的人CD4阳性CD25高表达Treg就可达到非常高的同种异体抗原特异性程度。细胞数量得到了有效增加。初次同种异体刺激似乎是产生同种异体抗原特异性Treg的一个先决条件，而用抗CD3加抗CD28单克隆抗体进行二次同种异体或多克隆刺激在相似程度上富集了同种异体抗原特异性和细胞产量。
结论/意义：我们所描述的体外扩增方案很可能会提高基于Treg的临床免疫疗法的成功率，并有助于诱导对选定抗原的耐受性，同时将一般免疫抑制降至最低。这种方法对于HLA不匹配移植的受者尤其有意义。
