SARS coronavirus nucleocapsid immunodominant T-cell epitope cluster is common to both exogenous recombinant and endogenous DNA-encoded immunogens.
SARS coronavirus nucleocapsid immunodominant T-cell epitope cluster is common to both exogenous recombinant and endogenous DNA-encoded immunogens.
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DOI：
10.1016/j.virol.2005.11.042
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发表时间：
2006-03-30
期刊：
影响因子：
3.7
通讯作者：
August TJ
中科院分区：
文献类型：
--
作者：
Gupta V;Tabiin TM;Sun K;Chandrasekaran A;Anwar A;Yang K;Chikhlikar P;Salmon J;Brusic V;Marques ET;Kellathur SN;August TJ
疫苗免疫原的T细胞表位应答与病原体抗原的T细胞表位应答之间的对应对于疫苗效力至关重要。在本研究中，我们分析了小鼠对三种不同形式的SARS冠状病毒核衣壳（N）的免疫应答谱：（1）外源性重组蛋白（N-GST）与弗氏佐剂;（2）编码未修饰的N的DNA作为内源性胞质蛋白（pN）;（3）编码N的DNA作为靶向溶酶体MHC II区室的LAMP-1嵌合体（p-LAMP-N）。共聚焦显微镜和免疫电子显微镜都记录了转染细胞中LAMP/N嵌合体的溶酶体运输。免疫小鼠的反应明显不同。最强的T细胞IFN-γ和CTL应答是对LAMP-N嵌合体，其次是pN免疫原。相比之下，N-GST引起强的T细胞IL-4，但最小的IFN-γ应答和更大的抗体应答。然而，尽管存在这些差异，对三种免疫原中的每一种的免疫显性T细胞ELISpot应答都是由相同的N肽引起的，其中最大的应答是由一簇五种重叠肽N76-114产生的，其中每一种肽含有对I类和除N76-93外的II类等位基因都具有高结合分数的九聚体H2 d结合结构域。这些结果表明，N的加工和呈递，无论是外源性的还是内源性的，导致共同的免疫显性表位，支持修饰的抗原递送和运输形式的有用性，特别是LAMP嵌合体作为候选疫苗。然而，T细胞反应的概况明显不同。对N蛋白加佐剂的显著Th-2和体液应答与对LAMP-N嵌合体的平衡IFN-γ和IL-4应答和强记忆CTL应答形成对比。
Correspondence between the T-cell epitope responses of vaccine immunogens and those of pathogen antigens is critical to vaccine efficacy. In the present study, we analyzed the spectrum of immune responses of mice to three different forms of the SARS coronavirus nucleocapsid (N): (1) exogenous recombinant protein (N-GST) with Freund's adjuvant; (2) DNA encoding unmodified N as an endogenous cytoplasmic protein (pN); and (3) DNA encoding N as a LAMP-1 chimera targeted to the lysosomal MHC II compartment (p-LAMP-N). Lysosomal trafficking of the LAMP/N chimera in transfected cells was documented by both confocal and immunoelectron microscopy. The responses of the immunized mice differed markedly. The strongest T-cell IFN-γ and CTL responses were to the LAMP-N chimera followed by the pN immunogen. In contrast, N-GST elicited strong T cell IL-4 but minimal IFN-γ responses and a much greater antibody response. Despite these differences, however, the immunodominant T-cell ELISpot responses to each of the three immunogens were elicited by the same N peptides, with the greatest responses being generated by a cluster of five overlapping peptides, N76–114, each of which contained nonameric H2d binding domains with high binding scores for both class I and, except for N76–93, class II alleles. These results demonstrate that processing and presentation of N, whether exogenously or endogenously derived, resulted in common immunodominant epitopes, supporting the usefulness of modified antigen delivery and trafficking forms and, in particular, LAMP chimeras as vaccine candidates. Nevertheless, the profiles of T-cell responses were distinctly different. The pronounced Th-2 and humoral response to N protein plus adjuvant are in contrast to the balanced IFN-γ and IL-4 responses and strong memory CTL responses to the LAMP-N chimera.
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