Kwon Minsuk, Choi Young Joon, Sa Moa, Park Su-Hyung, Shin Eui-Cheol
Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon 34141, Korea.
Biomedical Science and Engineering Interdisciplinary Program, Korea Advanced Institute of Science and Technology, Daejeon 34141, Korea.
Immune Netw. 2018 Dec 19;18(6):e45. doi: 10.4110/in.2018.18.e45. eCollection 2018 Dec.
Immune checkpoint inhibitors (ICIs), such as anti-PD-1 and anti-PD-L1 Abs, have shown efficacy for the treatment of various cancers. Although research has actively sought to develop new ICIs and immunomodulators, no efficient assay system is available to evaluate their functional activities. In the present study, we established a two-round MLR with human PBMCs for evaluation of the T cell-activating capacity of anti-PD-1 and other immunomodulators. We initially performed conventional MLR for this purpose. However, anti-PD-1 blocking Abs could not increase the proliferation of allo-reactive T cells in conventional MLR because PD-L1 and PD-L2 cells disappeared gradually during MLR. Therefore, we re-applied the same stimulator PBMCs to the allo-stimulated responder cells as a second-round MLR on day 6 when anti-PD-1 or immunomodulators were also added. In this two-round MLR, the proliferation of allo-reactive T cells was enhanced by anti-PD-1 in a dose-dependent manner or by immunomodulators, such as lenalidomide and galunisertib, a TGF-β receptor-1 inhibitor. Proliferation was further increased by the combination of immunomodulators with anti-PD-1. Here, we established a modified two-round MLR method with human PBMCs for evaluation of the functional activities of anti-PD-1 and immunomodulators.
免疫检查点抑制剂（ICIs），如抗PD-1和抗PD-L1抗体，已显示出对各种癌症的治疗效果。尽管研究一直在积极寻求开发新的ICIs和免疫调节剂，但目前尚无有效的检测系统来评估它们的功能活性。在本研究中，我们建立了一种用人外周血单核细胞（PBMCs）进行的两轮混合淋巴细胞反应（MLR），以评估抗PD-1和其他免疫调节剂的T细胞激活能力。我们最初为此目的进行了传统的MLR。然而，抗PD-1阻断抗体在传统MLR中不能增加同种异体反应性T细胞的增殖，因为在MLR过程中PD-L1和PD-L2细胞逐渐消失。因此，在第6天，当加入抗PD-1或免疫调节剂时，我们将相同的刺激PBMCs重新应用于同种异体刺激的反应细胞，作为第二轮MLR。在这种两轮MLR中，抗PD-1以剂量依赖的方式增强同种异体反应性T细胞的增殖，或由免疫调节剂如来那度胺和TGF-β受体-1抑制剂加鲁尼西替增强。免疫调节剂与抗PD-1联合使用可进一步增加增殖。在此，我们建立了一种用人PBMCs的改良两轮MLR方法，用于评估抗PD-1和免疫调节剂的功能活性。
