Ramachandra Lakshmi, Boom W Henry, Harding Clifford V
Department of Pediatrics, Division of Infectious Diseases, Case Western Reserve University and Rainbow Babies and Children's Hospital University, Cleveland, OH, USA.
Methods Mol Biol. 2008;445:353-77. doi: 10.1007/978-1-59745-157-4_23.
Phagocytic antigen-presenting cells (APCs) are involved in innate and adaptive immune responses to bacteria. Adaptive responses to bacteria involve processing of bacterial antigens for presentation by class II major histocompatibility complex (MHC II) molecules and class I MHC (MHC I) molecules to stimulate CD4(+) and CD8(+) T cells, respectively. To examine the role of phagosomes in processing of antigens for presentation by MHC II molecules to CD4(+) T cells, phagosomes have been biochemically and functionally analyzed by a variety of techniques that include flow analysis (flow organellometry), SDS-PAGE/Western blotting, and an antigen-presenting organelle assay. Using these techniques, we have demonstrated that phagosomes containing latex beads or Mycobacterium tuberculosis (MTB) contain components of the MHC II processing pathway and support the formation of peptide-MHC II complexes.
吞噬性抗原呈递细胞(APC)参与对细菌的固有免疫和适应性免疫反应。对细菌的适应性反应涉及处理细菌抗原,以便分别通过II类主要组织相容性复合体(MHC II)分子和I类MHC(MHC I)分子呈递,从而刺激CD4(+)和CD8(+) T细胞。为了研究吞噬体在处理抗原以供MHC II分子呈递给CD4(+) T细胞过程中的作用,已通过多种技术对吞噬体进行了生物化学和功能分析,这些技术包括流式分析(流式细胞器测定法)、SDS-PAGE/蛋白质免疫印迹法以及抗原呈递细胞器测定法。使用这些技术,我们已经证明,含有乳胶珠或结核分枝杆菌(MTB)的吞噬体含有MHC II加工途径的成分,并支持肽-MHC II复合物的形成。