PO.IM01.10 · 免疫学
由单克隆抗体包被的靶细胞活化的NK细胞可增强双特异性抗体诱导的T细胞增殖、活化和细胞毒性
NK cells activated by monoclonal antibody-coated target cells enhance bispecific antibody-induced T cell proliferation, activation and cytotoxicity
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：我们此前证明，T细胞辅助可增强由单克隆抗体（mAb）包被的靶细胞所活化的自然杀伤（NK）细胞的活力和细胞毒潜能。在此，我们评估反之是否也成立，即由mAb包被的NK细胞是否能增强双特异性抗体（bsAb）重定向T细胞的增殖、活化和细胞毒潜能。
方法：将Raji淋巴瘤细胞与去除了NK细胞的正常供者外周血单个核细胞以5:1的效靶比混合。以受控浓度（占总效应细胞的0%、5%或20%）加回自体NK细胞。共培养物用抗CD3×CD19双特异性抗体（blinatumomab）、抗CD20单克隆抗体（rituximab）或二者进行处理。在第2天和第4天更换培养基和抗体。5天后，评估T细胞增殖（Ki67）和细胞毒分子（包括granzyme B）的产生，同时以剩余Raji细胞数作为细胞毒性的量度。
结果：当存在较多数量的NK细胞时，blinatumomab与rituximab联合处理可增强T细胞增殖和活化。NK细胞的存在也增强了对Raji细胞的清除。NK细胞对单药blinatumomab或rituximab所致T细胞增殖和活化的影响较不明显。其他mAb与bsAb联合组合也观察到类似结果。
结论：由mAb包被的肿瘤细胞所活化的NK细胞可促进bsAb重定向T细胞的增殖、活化和细胞毒能力。这些数据提供了证据，表明由mAb活化的NK细胞与由bsAb活化的T细胞具有协同作用并相互提供交叉辅助。这为评估涉及同时给予mAb和bsAb的治疗策略提供了额外依据，此类策略可实现对NK细胞（通过mAb）和T细胞（通过bsAb）的同时共同衔接。T细胞数（×10³）无抗体 Rituximab Blinatumomab Rituximab与Blinatumomab；0% NK 54±13 57±16 56±62 77±74；5% NK 56±20 68±34 60±64 153±137；20% NK 67±39 92±34 81±86 212±147。T细胞Granzyme B；0% NK 96±204 128±351 2328±1534 3660±1277；5% NK 265±500 1412±834 4508±2754 11409±5077；20% NK 1043±1491 6882±2830 10876±3252 27083±9834。剩余Raji细胞（×10³）；0% NK 314±72 230±82 184±26 145±27；5% NK 259±84 93±80 174±20 50±79；20% NK 241±68 21±35 124±11 9±15。
查看英文原文 English abstract
Background: We previously demonstrated that T cell help can enhance the viability and cytotoxic potential of Natural Killer (NK) cells activated by monoclonal antibody (mAb)-coated target cells. Here we evaluated whether the reverse is also true, and that NK cells coated by mAb can enhance proliferation, activation, and the cytotoxic potential of bispecific antibody (bsAb) retargeted T cells.
Methods: Raji lymphoma cells and normal donor peripheral blood mononuclear cells depleted of NK cells were mixed at an effector to target ratio of 5:1. Autologous NK cells were added back in controlled concentrations (0%, 5% or 20% of total effector cells). Co-cultures were treated with anti-CD3×CD19 bispecific antibody (blinatumomab), anti-CD20 monoclonal antibody (rituximab) or both. Media and antibodies were refreshed on days 2 and 4. After 5 days, T cell proliferation (Ki67) and production of cytotoxic molecules (including granzyme B) was assessed as was the number of remaining Raji cells as a measure of cytotoxicity.
Results: Treatment with the combination of blinatumomab and rituximab enhanced T cell proliferation and activation when larger numbers of NK cells were present. The presence of NK cells also enhanced elimination of Raji cells. The impact of NK cells on T cell proliferation and activation in response to single agent blinatumomab or rituximab was less pronounced. Similar results were seen with other mAb and bsAb combinations.
Conclusion: NK cells, activated by mAb-coated tumor cells, promote the proliferation, activation and cytotoxic capacity of bsAb-retargeted T cells. These data provide evidence that NK cells activated by mAb and T cells activated by bsAb are synergistic and provide cross help to each other. This provides additional rationale for evaluating therapeutic strategies involving concurrent administration of mAb and bsAb that allows for simultaneous co-engagement of NK cells (via mAbs) and T cells (via bsAbs). Number T cells (×10 3 ) No Ab Rituximab Blinatumomab
Rituximab & Blinatumomab 0% NK 54±13 57±16 56±62 77±74 5% NK 56±20 68±34 60±64 153±137 20% NK 67±39 92±34 81±86 212±147 T cell Granzyme B 0% NK 96±204 128±351 2328±1534 3660±1277 5% NK 265±500 1412±834 4508±2754 11409±5077 20% NK 1043±1491 6882±2830 10876±3252 27083±9834 Remaining Raji cells (×10 3 ) 0% NK 314±72 230±82 184±26 145±27 5% NK 259±84 93±80 174±20 50±79 20% NK 241±68 21±35 124±11 9±15
利益披露 Disclosure
R. Amani, None..
J. Arora, None..
G. J. Weiner, None.