Pogonka T, Brattig N, Nde P N, Adam R, Seeber F, Zipfel P F, Titanji V P, Lucius R
Lehrstuhl für Molekulare Parasitologie, Humboldt-Universität zu Berlin, Germany.
Clin Immunol Immunopathol. 1997 Oct;85(1):56-66. doi: 10.1006/clin.1997.4400.
The localization of T and B cell epitopes on a well characterized 33-kDa protein of the filarial nematode Onchocerca volvulus (Ov33) was studied using peripheral blood mononuclear cells (PBMC) and sera from a total of 52 onchocerciasis patients with the generalized form of infection. A proportion of the PBMC samples proliferated in response to recombinant Ov33-GST fusion protein and to fusion free Ov33-6xHis. Proliferative responses of patient PBMC to seven truncated Ov33-6xHis polypeptides and to three synthetic peptides revealed at least one major and two minor T cell epitopes in the protein. The dominant T cell stimulating domain was localized between amino acids 113 and 143. ELISA studies with the Ov33-GST fusion protein revealed that patient sera contained Ov33-specific IgG1, IgG4, IgE, and IgM antibodies. Analysis of the IgG4 response with 10 truncated Ov33 polypeptides identified four B cell stimulating domains in the N-terminal, central, and C-terminal region of the molecule. The B cell domain recognized by the majority of sera was localized between amino acids 113 and 143. The data indicate that this region of the protein is the major T and B cell stimulating domain of Ov33 and might be relevant for vaccine development and for improved immunodiagnosis of onchocerciasis.
利用来自52例全身性盘尾丝虫病感染患者的外周血单个核细胞(PBMC)和血清,研究了盘尾丝虫33 kDa特征明确蛋白(Ov33)上T细胞和B细胞表位的定位。一部分PBMC样本对重组Ov33 - GST融合蛋白和无融合的Ov33 - 6xHis产生增殖反应。患者PBMC对7种截短的Ov33 - 6xHis多肽和3种合成肽的增殖反应显示该蛋白中至少有1个主要T细胞表位和2个次要T细胞表位。主要的T细胞刺激域定位于氨基酸113和143之间。用Ov33 - GST融合蛋白进行的ELISA研究表明,患者血清中含有Ov33特异性IgG1、IgG4、IgE和IgM抗体。用10种截短的Ov33多肽分析IgG4反应,在该分子的N端、中央和C端区域鉴定出4个B细胞刺激域。大多数血清识别的B细胞域定位于氨基酸113和143之间。数据表明,该蛋白的这一区域是Ov33主要的T细胞和B细胞刺激域,可能与疫苗开发及改进盘尾丝虫病的免疫诊断有关。