Terrén Iñigo, Astarloa-Pando Gabirel, Amarilla-Irusta Ainhoa, Borrego Francisco
Biocruces Bizkaia Health Research Institute, Immunopathology Group, Barakaldo, Spain.
Biocruces Bizkaia Health Research Institute, Immunopathology Group, Barakaldo, Spain; Ikerbasque, Basque Foundation for Science, Bilbao, Spain.
Methods Cell Biol. 2023;173:33-48. doi: 10.1016/bs.mcb.2022.02.002. Epub 2022 Mar 15.
Natural killer (NK) cells are part of the innate immune system, the classic cytotoxic population of innate lymphoid cells (ILCs). They can directly kill virus-infected or tumor cells through different mechanisms without prior sensitization using their lytic functions in response to different signals (target cell ligands and/or inflammatory cytokines) and secreting cytokines, such as interferon gamma (IFNγ) and tumor necrosis factor (TNF). NK cells use antibody-dependent cell-mediated cytotoxicity (ADCC) to recognize and kill cells expressing target antigens when they are antibody coated. Redirected cytotoxicity is a technique used to target cells that do not per se activate NK cells. Here, we use redirected degranulation, a surrogate technique that correlates with redirected lysis. The P815 cell line (mouse mastocytoma) express fragment crystallizable gamma receptor II (FcγRII) and therefore could bind the Fc portion of mouse IgG antibodies, which through their fragment antigen-binding (Fab) may recognize NK cells activating receptors leading to target cell lysis. This technique could be used to determine the inhibitory or activating capacity of different receptors or isoforms and in immunotherapy using T cell and NK cell activators.
自然杀伤（NK）细胞是先天性免疫系统的一部分，是先天性淋巴细胞（ILC）中的经典细胞毒性群体。它们可以通过不同机制直接杀伤病毒感染或肿瘤细胞，无需预先致敏，利用其裂解功能响应不同信号（靶细胞配体和/或炎性细胞因子）并分泌细胞因子，如干扰素γ（IFNγ）和肿瘤坏死因子（TNF）。当细胞被抗体包被时，NK细胞利用抗体依赖性细胞介导的细胞毒性（ADCC）识别并杀伤表达靶抗原的细胞。重定向细胞毒性是一种用于靶向本身不会激活NK细胞的细胞的技术。在这里，我们使用重定向脱颗粒，这是一种与重定向裂解相关的替代技术。P815细胞系（小鼠肥大细胞瘤）表达可结晶片段γ受体II（FcγRII），因此可以结合小鼠IgG抗体的Fc部分，这些抗体通过其抗原结合片段（Fab）可以识别NK细胞激活受体，导致靶细胞裂解。该技术可用于确定不同受体或异构体的抑制或激活能力，以及用于使用T细胞和NK细胞激活剂的免疫治疗。
