Paxton W A, Yazdanbakhsh M, Kurniawan A, Partono F, Maizels R M, Selkirk M E
Department of Biochemistry, Wellcome Centre for Parasitic Infections, Imperial College of Science, Technology and Medicine, London, United Kingdom.
Infect Immun. 1993 Jul;61(7):2827-33. doi: 10.1128/iai.61.7.2827-2833.1993.
We have isolated and sequenced clones encoding the repeated subunit of the surface-associated glycoprotein gp15/400 from the two nematode species predominantly responsible for lymphatic filariasis in humans: Brugia malayi and Wuchereria bancrofti. The amino acid sequence of the 15-kDa subunit, derived from the nucleotide sequence of the gene fragment from B. malayi, is identical to that previously reported for B. pahangi, whereas the derived W. bancrofti protein sequence differs in only 7 of 132 residues. The identity of the protein in the two Brugia species allowed us to use a recombinant from B. pahangi to examine the serological response of adult Indonesian subjects infected with B. malayi. The polymerase chain reaction-amplified subunit was expressed in Escherichia coli via the pDS56/RBS11 plasmid and purified by nickel-chelating chromatography. A significant proportion of individuals produced antigen-specific immunoglobulin E (IgE). This was most pronounced in the individuals with elephantiasis, with 14 of 15 showing elevated titers and a mean of 3.2 ng of specific IgE ml-1. Only 2 of 15 microfilaremic individuals possessed elevated titers of specific IgE, with a mean of 0.045 ng ml-1 for the group as a whole. Asymptomatic amicrofilaremic residents showed approximately equal numbers of responders (defined as having a value in the radioimmunoassay greater than two standard deviations above controls) and nonresponders, with a group mean of 1.2 ng of antigen-specific IgE ml-1.
我们已经从两种主要导致人类淋巴丝虫病的线虫中分离并测序了编码表面相关糖蛋白gp15/400重复亚基的克隆:马来布鲁线虫和班氏吴策线虫。从马来布鲁线虫基因片段的核苷酸序列推导而来的15 kDa亚基的氨基酸序列,与之前报道的彭亨布鲁线虫的序列相同,而推导得到的班氏吴策线虫蛋白质序列在132个残基中只有7个不同。两种布鲁线虫中该蛋白质的一致性使我们能够使用来自彭亨布鲁线虫的重组体来检测感染马来布鲁线虫的成年印度尼西亚受试者的血清学反应。通过聚合酶链反应扩增的亚基通过pDS56/RBS11质粒在大肠杆菌中表达,并通过镍螯合色谱法纯化。相当一部分个体产生了抗原特异性免疫球蛋白E(IgE)。这在患有象皮病的个体中最为明显,15名中有14名显示滴度升高,特异性IgE的平均水平为3.2 ng/ml。15名微丝蚴血症个体中只有2名特异性IgE滴度升高,整个组的平均水平为0.045 ng/ml。无症状无微丝蚴血症居民中反应者(定义为放射免疫测定值比对照高两个标准差以上)和无反应者的数量大致相等,组平均抗原特异性IgE水平为1.2 ng/ml。