Chatterjee B P, Santra A, Karmakar P R, Mazumder D N
Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Calcutta, India.
Trop Med Int Health. 1996 Oct;1(5):633-9. doi: 10.1111/j.1365-3156.1996.tb00088.x.
The excretory/secretory (ES) antigen(s) of Ascaris lumbricoides was fractionated into 10 fractions by gel chromatography on a Suparose 12 column in FPLC. Of these, the third fraction (Al III), showing binding activity with both IgE and IgG antibodies of A. lumbricoides infected patients' sera, was further resolved into 2 fractions (Al IIIa and Al IIIb) on passage through a Mono Q column. Al IIIb was found to be the most potent antigen due to its high binding affinity with IgE and IgG antibodies of Ascaris infected patients as evidenced by ELISA inhibition. Although a two to five-fold increase of serum IgE level was observed in all helminthic parasite infected patients studied compared to control subjects, Al IIIb specific IgE was detected in sera of all Ascaris infected and only 40% of hookworm infected patients. When Al IIIb was tested by ELISA with sera of control subjects, Ascaris, hookworm, Strongyloides and Trichuris infected patients, strong binding was observed with the IgE and also IgG of all the Ascaris infected patients; however, it cross-reacted with IgG in 50% of hookworm, 28.6% of Trichuris trichura and 22.2% of Strongyloides infected patients' sera; but with IgE only in 40% of hookworm infected patients' sera. Further study showed specific detection of IgG4 in all the serum samples of 65 Ascaris infected patients when Al IIIb antigen was allowed to react with different subclasses of IgG by ELISA, giving a sensitivity of 100%. Reactivities of Al IIIb with IgG1 and IgG3 were only 47.6 and 11.8% respectively and there was no reactivity with IgG2 subclass. No IgG4 reactivity against Al IIIb was observed in the sera of hookworm, Trichuris or Strongyloides infected patients and was similar to that observed with control subjects showing the 100% specificity of the test system. This study may therefore be regarded as a novel technique for serodiagnosis of ascariasis by measuring Ascaris specific IgG4.
通过在快速蛋白质液相色谱（FPLC）中使用Superose 12柱进行凝胶色谱法，将蛔虫的排泄/分泌（ES）抗原分离为10个组分。其中，第三组分（Al III）与蛔虫感染患者血清中的IgE和IgG抗体均表现出结合活性，在通过Mono Q柱后进一步分离为2个组分（Al IIIa和Al IIIb）。通过ELISA抑制试验证明，Al IIIb因其与蛔虫感染患者的IgE和IgG抗体具有高结合亲和力，而被发现是最有效的抗原。尽管与对照组相比，在所有研究的蠕虫寄生虫感染患者中均观察到血清IgE水平有两到五倍的升高，但仅在所有蛔虫感染患者的血清以及40%的钩虫感染患者的血清中检测到Al IIIb特异性IgE。当用对照组、蛔虫、钩虫、粪类圆线虫和鞭虫感染患者的血清通过ELISA检测Al IIIb时，在所有蛔虫感染患者的IgE和IgG中均观察到强结合；然而，它与50%的钩虫、28.6%的 Trichuris trichura和22.2%的粪类圆线虫感染患者的血清中的IgG发生交叉反应；但仅在40%的钩虫感染患者的血清中与IgE发生反应。进一步研究表明，当通过ELISA使Al IIIb抗原与IgG的不同亚类反应时，在65例蛔虫感染患者的所有血清样本中均特异性检测到IgG4，灵敏度为100%。Al IIIb与IgG1和IgG3的反应性分别仅为47.6%和11.8%，与IgG2亚类无反应性。在钩虫、鞭虫或粪类圆线虫感染患者的血清中未观察到针对Al IIIb的IgG4反应性，且与对照组观察到的情况相似，表明该检测系统具有100%的特异性。因此，本研究可被视为一种通过测量蛔虫特异性IgG4进行蛔虫病血清诊断的新技术。
