Vos G J, Gardiner P R
International Laboratory for Research on Animal Diseases, Nairobi, Kenya.
Parasitology. 1990 Feb;100 Pt 1:93-100. doi: 10.1017/s0031182000060157.
Sera from goats and cattle that were infected with two Trypanosoma vivax clones (ILDat 1.2 and ILDat 2.1) derived from different stocks were analysed for antibody activity against the variable surface glycoproteins (VSGs) of the infecting clones by enzyme-linked immune assays (ELISA) and immune lysis. To obtain purified VSG, lysed trypanosomes were separated on dodecyl sulphate-polyacrylamide gels. The gels were copper stained and the VSG protein band was excised from the gel. After destaining, the proteins were electroeluted from the gel slices and used as antigens in ELISA. High titres of IgM and IgG1 antibodies and lytic antibodies against the VSG of the infecting clone were detected. The IgG1 response appeared about 4 days later than the IgM response. IgG2 antibodies were only detected in goats and cattle that were infected with ILDat 1.2. Two goats and two calves that were infected with ILDat 1.2 showed recurrent peaks in lytic activity and of IgM and IgG1 antibody activity to the VSG of the infecting variable antigenic type (VAT). Two goats that were infected with ILDat 2.1 showed a similar pattern, but in two other goats there was a recurrent peak only in the IgM class. Recurrent peaks of antibody activity to the VSG of ILDat 1.2 and ILDat 2.1 were not detected in the sera of goats that had been inoculated with irradiated trypanosomes or that had been infected with an unrelated T. vivax clone. The recurrence of antibody peaks against the VSG of infecting VATs suggests that trypanosomes with completely or partially identical surface determinants reappear during T. vivax infection of ruminants.
对感染了源自不同毒株的两个间日疟原虫克隆(ILDat 1.2和ILDat 2.1)的山羊和牛的血清,通过酶联免疫测定(ELISA)和免疫裂解分析其针对感染克隆可变表面糖蛋白(VSG)的抗体活性。为获得纯化的VSG,将裂解的锥虫在十二烷基硫酸钠 - 聚丙烯酰胺凝胶上分离。凝胶进行铜染色,然后从凝胶中切下VSG蛋白条带。脱色后,从凝胶切片中电洗脱蛋白质,并用作ELISA中的抗原。检测到针对感染克隆VSG的高滴度IgM和IgG1抗体以及裂解抗体。IgG1反应比IgM反应出现约晚4天。仅在感染ILDat 1.2的山羊和牛中检测到IgG2抗体。两只感染ILDat 1.2的山羊和两只小牛对感染的可变抗原型(VAT)的VSG的裂解活性以及IgM和IgG1抗体活性出现反复峰值。两只感染ILDat 2.1的山羊表现出类似模式,但另外两只山羊仅在IgM类别中出现反复峰值。在接种了经辐照的锥虫或感染了无关间日疟原虫克隆的山羊血清中,未检测到针对ILDat 1.2和ILDat 2.1的VSG的抗体活性反复峰值。针对感染VAT的VSG的抗体峰值反复出现表明,在反刍动物间日疟原虫感染期间,具有完全或部分相同表面决定簇的锥虫会再次出现。