Gober Joshua G, Capietto Aude-Hélène, Hoshyar Reyhane, Darwish Martine, Vandlen Richard, Linehan Jonathan L, Delamarre Lélia, ElSohly Adel M
Department of Protein Chemistry, Genentech Inc, South San Francisco, CA, USA.
Cancer Immunology Department, Genentech Inc, South San Francisco, CA, USA.
iScience. 2025 Mar 13;28(4):112212. doi: 10.1016/j.isci.2025.112212. eCollection 2025 Apr 18.
Recent studies suggest that CD4 T cells can exert potent anti-tumor effects and improve immunotherapy efficacy by aiding CD8 T cells. However, characterizing the mechanism of CD4 T cells' anti-tumor activity has been challenging due to inaccurate major histocompatibility complex class II (MHC-II) peptide prediction algorithms and the lack of high-quality reagents for immune monitoring. To address this, we developed MHC2-substitution of CLIP and analytical LCMS evaluation (MHC2-SCALE), a streamlined approach combining affinity optimized class II-associated invariant chain peptide (CLIP) exchange technology, high throughput 2D-LCMS analysis, and rapid generation of peptide-bound MHC-II monomers for subsequent multimer assembly. We validated MHC-II peptide candidates predicted by the immune epitope database (IEDB) algorithm, as well as uncovered many true and immunogenic MHC-II binders that were not predicted by IEDB. Thus, MHC2-SCALE expands the opportunities for discovering, tracking, and phenotyping antigen-specific CD4 T cells in preclinical and clinical settings, thereby improving therapies for cancer, autoimmunity, or infectious diseases.
最近的研究表明，CD4 T细胞可通过辅助CD8 T细胞发挥强大的抗肿瘤作用并提高免疫治疗效果。然而，由于主要组织相容性复合体II类（MHC-II）肽预测算法不准确以及缺乏用于免疫监测的高质量试剂，表征CD4 T细胞抗肿瘤活性的机制一直具有挑战性。为了解决这一问题，我们开发了CLIP的MHC2替代和分析LCMS评估（MHC2-SCALE），这是一种简化方法，结合了亲和力优化的II类相关恒定链肽（CLIP）交换技术、高通量二维LCMS分析以及快速生成肽结合的MHC-II单体以用于后续多聚体组装。我们验证了免疫表位数据库（IEDB）算法预测的MHC-II肽候选物，并发现了许多IEDB未预测到的真实且具有免疫原性的MHC-II结合物。因此，MHC2-SCALE扩展了在临床前和临床环境中发现、追踪和鉴定抗原特异性CD4 T细胞的机会，从而改善癌症、自身免疫性疾病或传染病的治疗方法。
