Antigenic peptides play a central role in immune surveillance in cancer, infectious disease, autoimmunity, and allergy. The identification and isolation of antigenic peptides for T cell immune response are crucial for successful personalized adoptive immune cell therapy. The mainly methods includes gene sequencing and bioinformatic analysis. The antigenic peptides which identified by analysis and artificially synthesized still need antigen presenting cell (APC) to deliver to T cells. However, high costs and lengthy process times have limited its application in clinical practice. In order to overcome it, this study attempted to directly capture antigenic peptide-major histocompatibility complex (MHC) class I (pMHCs) from cell lysates using streptavidin Dynabeads and biotin-labeled antibodies, then the pMHCs was co-cultured with tumor infiltrating lymphocytes (TILs) of the same tissue origin. The results indicated that the captured pMHCs were able to enrich the tumor antigen-specific CD8⁺ T cells, and also effectively induce proliferation and cytotoxic responses of CD8⁺ T cells. This study provided a novel approach for obtaining tumor antigenic pMHCs, which could enrich antigen-specific CD8⁺ T cells, and could also function as artificial APCs (aAPCs) to stimulate proliferation and activation of T cells. Notably, these pMHCs can stimulate the proliferation of stem-like memory T cells. In conclusion, this study describes a time-saving and low-cost method to isolate tumour antigen peptide MHC complexs, helping tumor antigen-specific T cell enrichment, activation, and proliferation.

抗原肽在癌症、传染病、自身免疫和过敏的免疫监视中发挥着核心作用。T 细胞免疫反应的抗原肽的鉴定和分离对于成功的个性化过继免疫细胞治疗至关重要。主要方法包括基因测序和生物信息分析。通过分析鉴定并人工合成的抗原肽仍需要抗原呈递细胞（APC）来递送至T细胞。然而，高昂的成本和漫长的流程限制了其在临床实践中的应用。为了克服这个问题，本研究尝试使用链霉亲和素Dynabeads和生物素标记抗体从细胞裂解物中直接捕获抗原肽主要组织相容性复合物（MHC）I类（pMHC），然后将pMHC与肿瘤浸润淋巴细胞共培养。 TIL）具有相同的组织来源。结果表明，捕获的pMHC能够富集肿瘤抗原特异性CD8 ⁺ T细胞，并有效诱导CD8 ⁺ T细胞的增殖和细胞毒反应。这项研究提供了一种获得肿瘤抗原pMHC的新方法，它可以富集抗原特异性CD8 ⁺ T细胞，并且还可以作为人工APC（aAPC）来刺激T细胞的增殖和活化。值得注意的是，这些 pMHC 可以刺激干细胞样记忆 T 细胞的增殖。总之，本研究描述了一种省时、低成本的分离肿瘤抗原肽MHC复合物的方法，有助于肿瘤抗原特异性T细胞的富集、激活和增殖。