Nehete P N, Gambhira R, Nehete B P, Sastry K Jagannadha
Departments of Veterinary Sciences and Molecular Pathology, M.D. Anderson Cancer Center, The University of Texas, 650 Cool Water Drive, Bastrop, TX 78602, USA.
J Med Primatol. 2003 Apr;32(2):67-73. doi: 10.1034/j.1600-0684.2003.00011.x.
Detection and enumeration of functional antigen-specific T cells is important for understanding the breadth of cell-mediated immunity to infections and experimental vaccines. We tested the utility of dendritic cells (DC), the professional antigen presenting cells, in the enzyme-linked immunosorbent spot-forming cell assay (ELISPOT) for efficient monitoring of antigen-specific immunity in rhesus macaques vaccinated with an HIV envelope peptide-cocktail. Compared with direct antigen-specific stimulation of peripheral blood mononuclear cells, the DC-ELISPOT protocol involving co-culturing of macaque T cells with autologous DC pulsed with the various peptides from the vaccine cocktail yielded up to 18-fold higher numbers of interferon-gamma producing cells without increasing the background. Importantly, use of DC in the analyses revealed immune responses in vaccinated macaques that were otherwise undetectable. Similar data were obtained when recall responses to purified protein derivative were analyzed by the DC-ELISPOT method using blood samples from human volunteers. These data establish the importance of DC in improving detection sensitivity and eliminating false negative results, both essential for efficient monitoring of antigen-specific cellular immune responses.
检测和计数功能性抗原特异性T细胞对于理解细胞介导的抗感染免疫及实验疫苗免疫的广度至关重要。我们测试了树突状细胞（DC）（专业抗原呈递细胞）在酶联免疫斑点形成细胞测定（ELISPOT）中用于有效监测接种HIV包膜肽鸡尾酒的恒河猴抗原特异性免疫的效用。与外周血单核细胞的直接抗原特异性刺激相比，DC-ELISPOT方案（涉及将猕猴T细胞与用疫苗鸡尾酒中的各种肽脉冲处理的自体DC共培养）产生的产生γ干扰素的细胞数量增加了18倍，且未增加背景。重要的是，在分析中使用DC揭示了接种疫苗的猕猴中原本无法检测到的免疫反应。当使用来自人类志愿者的血样通过DC-ELISPOT方法分析对纯化蛋白衍生物的回忆反应时，也获得了类似的数据。这些数据证明了DC在提高检测灵敏度和消除假阴性结果方面的重要性，这两者对于有效监测抗原特异性细胞免疫反应都是必不可少的。
