Erttmann Klaus D, Kleensang André, Schneider Erik, Hammerschmidt Sven, Büttner Dietrich W, Gallin Michaela
Bernhard Nocht Institute for Tropical Medecine, Bernhard-Nocht-Str.74, D-20359 Hamburg, Germany.
Biochim Biophys Acta. 2005 Jun 30;1741(1-2):85-94. doi: 10.1016/j.bbadis.2004.12.010. Epub 2005 Jan 5.
In the search for Onchocerca volvulus antigens possibly involved in protection against human onchocerciasis, partial amino acid sequence analysis of one of the O. volvulus antigens of the serologically identified proteins showed a close relationship to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein family. Subsequent adult worm cDNA library screening and cloning produced a clone of 1650 bp. An open reading frame spans over 1020 bp encoding for a protein of 340 amino acids with an apparent molecular weight of 38000. Comparison of the complete amino acid sequence identified this protein as a member of the GAPDH protein family. The recombinantly expressed protein shows GAPDH enzymatic activity as well as plasminogen-binding capacity. DNA sequence analysis of the corresponding gene revealed the presence of two introns. Using immunohistology Ov-GAPDH was observed in microfilariae, infective larvae, and adult male and female worms. Most striking was the labelling of the musculature of the body wall. Labelling was also observed in the pseudocoeloma cavity and in a subset of cell nuclei, suggesting additional, non-glycolytic functions of the Ov-GAPDH. Gene gun immunization with the DNA-construct in cattle led to specific humoral immune responses. Thus, the protective potential of the DNA-construct of Ov-GAPDH can be evaluated in vaccination trials using animal models such as the cattle/Onchocerca ochengi model.
在寻找可能参与抵御人类盘尾丝虫病的盘尾丝虫抗原的过程中，对血清学鉴定蛋白质中一种盘尾丝虫抗原进行的部分氨基酸序列分析显示，它与甘油醛-3-磷酸脱氢酶（GAPDH）蛋白家族关系密切。随后对成虫cDNA文库进行筛选和克隆，得到了一个1650 bp的克隆。一个开放阅读框跨越1020 bp，编码一个由340个氨基酸组成的蛋白质，其表观分子量为38000。完整氨基酸序列的比较确定该蛋白质为GAPDH蛋白家族的一员。重组表达的蛋白质显示出GAPDH酶活性以及纤溶酶原结合能力。相应基因的DNA序列分析揭示存在两个内含子。通过免疫组织学方法在微丝蚴、感染性幼虫以及成虫雌雄虫体中均观察到了盘尾丝虫GAPDH（Ov-GAPDH）。最显著的是体壁肌肉组织的标记。在假体腔以及一部分细胞核中也观察到了标记，这表明Ov-GAPDH具有额外的非糖酵解功能。用该DNA构建体对牛进行基因枪免疫可引发特异性体液免疫反应。因此，可在使用牛/奥氏盘尾丝虫模型等动物模型的疫苗试验中评估Ov-GAPDH DNA构建体的保护潜力。
