肿瘤细胞治疗研究
英文原题：High yield killing of lymphoma cells by anti-CD22 CAR-NK cell therapy.
High yield killing of lymphoma cells by anti-CD22 CAR-NK cell therapy.
嵌合抗原受体 (CARs) 为通过免疫系统靶向 B 细胞恶性肿瘤提供了一种有前景的新方法。
嵌合抗原受体（CAR）为通过免疫系统靶向B细胞恶性肿瘤提供了有前景的新方法。尽管靶向CD19和CD22的CAR-T细胞已在血液系统恶性肿瘤中证实有效，其制备仍然高度复杂。与T细胞不同，NK细胞可不依赖抗原特异性清除靶细胞，同时避免移植物抗宿主病（GvHD）。CAR-NK细胞被认为比CAR-T细胞更安全，因为其生命周期较短且产生的毒性细胞因子较少。NK-92细胞在体外具有无限增殖能力，可作为制备CAR工程化NK细胞的来源。我们发现，由抗CD22单克隆抗体m971构建的CAR特异性靶向CD22近膜表位，其抗白血病活性优于采用其他结合结构域构建的CAR。为进一步增强NK细胞的抗白血病能力，我们采用慢病毒转导制备m971-CD28-CD3 NK-92细胞。CD22在B细胞淋巴瘤中高表达。为评估靶向CD22的潜力，我们选用Raji细胞作为CD22阳性细胞。本研究旨在探讨CD22作为CAR-NK-92疗法治疗B细胞淋巴瘤的潜在靶点。我们首先在体外制备并验证表达CD22结合CAR的m971-CD28-CD3 NK-92细胞，通过流式细胞术分析CAR表达；使用7AAD流式细胞术检测其对淋巴瘤靶细胞系的细胞毒性，并通过ELISA评估CAR-NK-92细胞接触靶细胞后的细胞因子生成。m971-CD28-CD3 NK-92细胞成功表达CD22特异性CAR，可有效裂解表达CD22的淋巴瘤细胞系；与靶细胞共培养后产生大量IFN-γ和GM-CSF等细胞因子，而IL-6水平较低。结果表明，转移m971-CD28-CD3 NK-92细胞有望成为治疗B细胞淋巴瘤的免疫疗法。
Chimeric antigen receptors (CARs) offer a promising new approach for targeting B cell malignancies through the immune system. Despite the proven effectiveness of CAR T cells targeting CD19 and CD22 in hematological malignancies, it is imperative to note that their production remains a highly complex process. Unlike T cells, NK cells eliminate targets in a non-antigen-specific manner while avoiding graft vs. host disease (GvHD). CAR-NK cells are considered safer than CAR-T cells because they have a shorter lifespan and produce less toxic cytokines. Due to their unlimited ability to proliferate in vitro, NK-92 cells can be used as a source for CAR-engineered NK cells. We found that CARs created from the m971 anti-CD22 mAb, which specifically targets a proximal CD22 epitope, were more effective at anti-leukemic activity compared to those made with other binding domains. To further enhance the anti-leukemic capacity of NK cells, we used lentiviral transduction to generate the m971-CD28-CD3 NK-92. CD22 is highly expressed in B cell lymphoma. To evaluate the potential of targeting CD22, Raji cells were selected as CD22-positive cells. Our study aimed to investigate CD22 as a potential target for CAR-NK-92 therapy in the treatment of B cell lymphoma. We first generated m971-CD28-CD3 NK-92 that expressed a CAR for binding CD22 in vitro. Flow cytometric analysis was used to evaluate the expression of CAR. The 7AAD determined the cytotoxicity of the m971-CD28-CD3 NK-92 towards target lymphoma cell lines by flow cytometry assay. The ELISA assay evaluated cytokine production in CAR NK-92 cells in response to target cells. The m971-CD28-CD3 NK-92 cells have successfully expressed the CD22-specific CAR. m971-CD28-CD3 NK-92 cells efficiently lysed CD22-expressing lymphoma cell lines and produced large amounts of cytokines such as IFN- and GM-CSF but a lower level of IL-6 after coculturing with target cells. Based on our results, it is evident that transferring m971-CD28-CD3 NK-92 cells could be a promising immunotherapy for B cell lymphoma.
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