Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad de México, México.
CONACyT-Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad de México, México.
Front Immunol. 2020 Oct 2;11:577815. doi: 10.3389/fimmu.2020.577815. eCollection 2020.
T cells recognizing epitopes on the surface of mycobacteria-infected macrophages can impart protection, but with associated risk for reactivation to lung pathology. We aimed to identify antibodies specific to such epitopes, which carry potentials for development toward novel therapeutic constructs. Since epitopes presented in the context of major histocompatibility complex alleles are rarely recognized by naturally produced antibodies, we used a phage display library for the identification of monoclonal human single domain antibody producing clones. The selected 2C clone displayed T cell receptor-like recognition of an HLA-A*0201 bound KLVANNTRL peptide from the Ag85B antigen, which is known to be an immunodominant epitope for human T cells. The specificity of the selected domain antibody was demonstrated by solid phase immunoassay and by immunofluorescent surface staining of peptide loaded cells of the T2 cell line. The antibody affinity binding was determined by biolayer interferometry. Our results validated the used technologies as suitable for the generation of antibodies against epitopes on the surface of infected cells. The potential approaches forward the development of antibody in immunotherapy of tuberculosis have been outlined in the discussion.
T 细胞识别分枝杆菌感染的巨噬细胞表面的表位可以提供保护，但也存在与肺病理学再激活相关的风险。我们旨在识别针对这些表位的抗体，这些抗体具有开发新型治疗性构建体的潜力。由于主要组织相容性复合体等位基因背景下呈现的表位很少被天然产生的抗体识别，因此我们使用噬菌体展示文库来鉴定产生单域抗体的单克隆人克隆。所选的 2C 克隆显示出对 HLA-A*0201 结合的 KLVANNTRL 肽的 T 细胞受体样识别，该肽是 Ag85B 抗原的免疫优势表位，已知是人类 T 细胞的免疫优势表位。所选的结构域抗体的特异性通过固相免疫测定和肽负载 T2 细胞系细胞的免疫荧光表面染色来证明。抗体亲和力结合通过生物层干涉测量法确定。我们的结果验证了所使用的技术适用于针对感染细胞表面表位产生抗体。讨论中概述了在结核病免疫治疗中开发抗体的潜在方法。
