Identification of vaccine candidate peptides in the NcSRS2 surface protein of Neospora caninum by using CD4+ cytotoxic T lymphocytes and gamma interferon-secreting T lymphocytes of infected holstein cattle.
Identification of vaccine candidate peptides in the NcSRS2 surface protein of Neospora caninum by using CD4+ cytotoxic T lymphocytes and gamma interferon-secreting T lymphocytes of infected holstein cattle.
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使用感染荷斯坦牛的 CD4 细胞毒性 T 淋巴细胞和γ干扰素分泌 T 淋巴细胞鉴定犬新孢子虫 NcSRS2 表面蛋白中的疫苗候选肽。
DOI：
10.1128/iai.73.3.1321-1329.2005
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发表时间：
2005
期刊：
影响因子：
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通讯作者：
Baszler,TimothyV
中科院分区：
文献类型：
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作者：
Staska,LaurenM;Davies,ChristopherJ;Brown,WendyC;McGuire,TravisC;Suarez,CarlosE;Park,JooYoun;Mathison,BruceA;Abbott,JeffreyR;Baszler,TimothyV
以前，我们的实验室表明，荷斯坦牛实验感染犬新孢子虫发展寄生虫特异性CD 4+细胞毒性T淋巴细胞（CTL），通过穿孔素-颗粒酶途径裂解感染的自体靶细胞。为了鉴定诱导牛CTL和辅助性T淋巴细胞应答的特异性寄生虫抗原以用于针对牛新孢子虫病的疫苗开发，靶向了速殖子主要表面蛋白NcSAG 1和NcSRS 2。在全速殖子抗原扩增的牛T淋巴细胞系中，重组NcSRS 2诱导了有效的记忆性CD 4+和CD 8 + T淋巴细胞活化，如增殖和γ干扰素（IFN-γ）分泌所示，而重组NcSAG 1诱导了最小的记忆反应。随后，通过使用覆盖整个NcSRS 2序列的重叠肽，对NcSRS 2的T淋巴细胞表位承载肽进行作图。四个实验感染牛与六个不同的主要组织相容性复合体（MHC）II类单倍型的免疫细胞的来源，用于识别NcSRS 2肽提出的荷斯坦MHC单倍型。通过使用rNcSRS 2刺激的短期T淋巴细胞系的IFN-γ分泌、外周血单个核细胞的IFN-γ酶联免疫斑点（ELISPOT）测定和rNcSRS 2刺激的效应细胞的51 Cr释放细胞毒性测定来定位NcSRS 2肽。FourN.犬感染的荷斯坦牛产生NcSRS 2肽特异性T淋巴细胞，通过IFN-γ ELISPOT在外周血中离体检测，并通过测量T淋巴细胞IFN-γ产生和细胞毒性在体外检测。NcSRS 2跨越氨基酸133至155的免疫显性区域被来自四头牛的CD 4 +T淋巴细胞识别。这些发现支持了亚单位N的研究。在不同MHC基因型牛中诱导NcSRS 2肽特异性CTL和IFN-γ分泌性T淋巴细胞的含有NcSRS 2基因序列或肽的犬疫苗。
Previously, our laboratory showed that Holstein cattle experimentally infected withNeospora caninumdevelop parasite-specific CD4+cytotoxic T lymphocytes (CTL) that lyse infected, autologous target cells through a perforin-granzyme pathway. To identify specific parasite antigens inducing bovine CTL and helper T-lymphocyte responses for vaccine development against bovine neosporosis, the tachyzoite major surface proteins NcSAG1 and NcSRS2 were targeted. In whole tachyzoite antigen-expanded bovine T-lymphocyte lines, recombinant NcSRS2 induced potent memory CD4+- and CD8+-T-lymphocyte activation, as indicated by proliferation and gamma interferon (IFN-γ) secretion, while recombinant NcSAG1 induced a minimal memory response. Subsequently, T-lymphocyte epitope-bearing peptides of NcSRS2 were mapped by using overlapping peptides covering the entire NcSRS2 sequence. Four experimentally infected cattle with six different major histocompatibility complex (MHC) class II haplotypes were the source of immune cells used to identify NcSRS2 peptides presented by Holstein MHC haplotypes. NcSRS2 peptides were mapped by using IFN-γ secretion by rNcSRS2-stimulated, short-term T-lymphocyte cell lines, IFN-γ enzyme-linked immunospot (ELISPOT) assay with peripheral blood mononuclear cells, and51Cr release cytotoxicity assay of rNcSRS2-stimulated effector cells. FourN. caninum-infected Holstein cattle developed NcSRS2 peptide-specific T lymphocytes detected ex vivo in peripheral blood by IFN-γ ELISPOT and in vitro by measuring T-lymphocyte IFN-γ production and cytotoxicity. An immunodominant region of NcSRS2 spanning amino acids 133 to 155 was recognized by CD4+T lymphocytes from the four cattle. These findings support investigation of subunitN. caninumvaccines incorporating NcSRS2 gene sequences or peptides for induction of NcSRS2 peptide-specific CTL and IFN-γ-secreting T lymphocytes in cattle with varied MHC genotypes.