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首页> 外文期刊>Infection and immunity >Identification of two Th1 cell epitopes on the Babesia bovis-encoded 77-kilodalton merozoite protein (Bb-1) by use of truncated recombinant fusion proteins.
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Identification of two Th1 cell epitopes on the Babesia bovis-encoded 77-kilodalton merozoite protein (Bb-1) by use of truncated recombinant fusion proteins.

机译：使用截短的重组融合蛋白鉴定牛肝菌编码的77-千达尔顿裂殖子蛋白（Bb-1）上的两个Th1细胞表位。

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Previous studies have demonstrated the serologic and T-cell immunogenicity for cattle of a recombinant form of the apical complex-associated 77-kDa merozite protein of Babesia bovis, designated Bb-1. The present study characterizes the immunogenic epitopes of the Bb-1 protein. A series of recombinant truncated fusion proteins spanning the majority of the Bb-1 protein were expressed in Escherichia coli, and their reactivities with bovine peripheral blood mononuclear cells and T-cell clones derived from B. bovis-immune cattle and with rabbit antibodies were determined. Lymphocytes from two immune cattle were preferentially stimulated by the N-terminal half of the Bb-1 protein (amino acids 23 to 266, termed Bb-1A), localizing the T-cell epitopes to the Bb-1A portion of the molecule. CD4+ T-cell clones derived by stimulation with the intact Bb-1 fusion protein were used to identify two T-cell epitopes in the Bb-1A protein, consisting of amino acids SVVLLSAFSGN VWANEAEVSQVVK and FSDVDKTKSTEKT (residues 23 to 46 and 82 to 94). In contrast, rabbit antiserum raised against the intact fusion protein reacted only with the C-terminal half of the protein (amino acids 267 to 499, termed Bb-1B), which contained 28 tandem repeats of the tetrapeptide PAEK or PAET. Biological assays and Northern (RNA) blot analyses for cytokines revealed that following activation with concanavalin A, T-cell clones reactive against the two Bb-1A epitopes produced interleukin-2, gamma interferon, and tumor necrosis factors beta and alpha, but not interleukin-4, suggesting that the Bb-1 antigen preferentially stimulates the Th1 subset of CD4+ T cells in cattle. The studies described here report for the first time the characterization, by cytokine production, of the Th1 subset of bovine T cells and show that, as in mice, protozoal antigens can induce Th1 cells in ruminants. This first demonstration of B. bovis-encoded Th1 cell epitopes provides a rationale for incorporation of all or part of the Bb-1 protein into a recombinant vaccine.
机译：先前的研究已经证明了牛复数种根尖菌复合体的77 kDa蛇纹石蛋白的重组形式对牛的血清学和T细胞免疫原性，命名为Bb-1。本研究表征了Bb-​​1蛋白的免疫原性表位。在大肠杆菌中表达了一系列跨越大部分Bb-1蛋白的重组截短融合蛋白，并确定了它们与牛外周血单核细胞和牛免疫牛原性牛的T细胞克隆以及兔抗体的反应性。 。 Bb-1蛋白的N末端一半（氨基酸23至266，称为Bb-1A）优先刺激来自两只免疫牛的淋巴细胞，从而将T细胞表位定位在分子的Bb-1A部分。通过完整Bb-1融合蛋白刺激获得的CD4 + T细胞克隆用于鉴定Bb-1A蛋白中的两个T细胞表位，由氨基酸SVVLLSAFSGN VWANEAEVSQVVK和FSDVDKTKSTEKT组成（残基23至46和82至94）。 。相反，针对完整融合蛋白的兔抗血清仅与该蛋白的C端一半反应（氨基酸267至499，称为Bb-1B），其中包含28个串联的四肽PAEK或PAET重复序列。针对细胞因子的生物学分析和Northern（RNA）印迹分析显示，在伴刀豆球蛋白A激活后，对两个Bb-1A表位有反应性的T细胞克隆产生白介素2，γ干扰素和肿瘤坏死因子β和α，但不产生白细胞介素。 -4，表明Bb-1抗原优先刺激牛CD4 + T细胞的Th1子集。此处描述的研究首次报道了通过细胞因子产生牛T细胞Th1亚型的特征，并表明，与小鼠一样，原生动物抗原可以在反刍动物中诱导Th1细胞。牛双歧杆菌编码的Th1细胞表位的第一个证明为将全部或部分Bb-1蛋白掺入重组疫苗提供了理论依据。

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