بررسي سرم بيماران مبتلا به واژنيت حاد و مزمن از نظر IgE و IgG اختصاص نسبت به كانديدا آلبيكانس با روش ايمونوبلاتينگ

Immunoblotting analysis of sera from patients with acute and chronic vaginitis for IgE and IgG antibiodies against Candida albicans

Background and purpose : Candida albicans is an environmental microflora of vagina. The circumstance of response to antigenic component of Candida albicans is variable in different patients, and probably plays an important role in course of vaginitis. The purpose of this study was to determine IgE and IgG antibodies to Candida albicans in sera from patients with acute and chronic candida and non-candidal vaginitis. Materials and methods : In this study, Candida albicans was grown on Medium. Then yeast cells were carefully harvested from the surface of the solid medium. After disruption of the yeast cells, samples were centrifuged at 25000 rpm. Supernatants were collected and used as a crude extract. Protein components of the crude extracts were separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and transferred to nitrocellulose filter. The component vaginitis (n-80, 20 patients from each groups) and visualized by Enzyme-labeled anti-human IgE and IgG antibodies in chromogenic substrate. Results : in SDS-PAGE a total of 15 different protein bands (13-75 Kda) were observed in crude extract of C.albicans, however in immunoblotting none of protein bands gave response with IgE antibodies. On the contrary, protein bands with molecular weight of 25 and 52 KDs gave strong response with IgG antibodies in 100% of sera from chronic candidal vaginitis patients. Whereas, acute non-candidal vaginitis patients did not react with any protein bands. 100% of sera from acute candidal vaginitis patients gave strong response only with 47 Kda protein bands and 55% of chronic non-candidal vaginitis patients gave weak response with 47 Kda protein bands. Conclusion : Our results showed that anti-Candida IgE to 25 Kda protein bands and 47 Kda protein band was strongly related to chronic and acute course of candidal vaginitis, respectively.