G.vaginalis'in sekiz epidemiyolojik tipinin imminodominant yapısal proteinleri ve adherans özelliklerinin belirlenmesi
Determination of imminodominant structural proteins and adherence properties of eight biotypes of gardnerella vaginalis
Yazar: FETHİYE KÖSEAHMET KOLAYLI
Danışman: PROF. DR. MURAT ERTÜRK
Karadeniz Teknik Üniversitesi / Sağlık Bilimleri Enstitüsü / Mikrobiyoloji Ana Bilim Dalı
Derece: Doktora, 1999
Konu: Mikrobiyoloji = Microbiology
Dizin: Blotting-western = Blotting-western ; Gardnerella vaginalis = Gardnerella vaginalis

Özet:
Bu çalışmada G.vaginalis'in sekiz epidemiyolojik biyotipinin SDS-PAGE ile protein profilleri çıkarılıp, antiserumlarla yapılan immünoblotlarda yapısal proteinlerin antijenik ve immunodominant özellikleri araştırıldı. Ayrıca herbir biyotipin vajinal epitel hücrelere adher olma kapasiteleri arasında fark olup olmadığı incelendi. Jel elektroforez profilleri değerlendirildiğinde biyotiplerin protein yoğunluktan ve sayılarında farklılıklar olduğu; ancak, bu farklılığın biyotipleri ayırt etmeye olanak sağlayacak düzeyde anlamlı olmadığı belirlendi. Western blot çalışmalarında, jel elektroforezinde açıkça belirlenemeyen, fakat antiserumlarla güçlü reaksiyon veren, MA'sı 41kDa proteinin diğer proteinlere göre immunodominant olduğu belirlendi. Antijenik yapı bakımından Biyotipler 1, 2 ve 5'in benzer reaksiyonlar verdiği; Biyotip 4, 7 ve 8'in antijenik özelliklerinin diğer biyotiplere oranla daha az olduğu belirlendi. Vajinal epitel hücrelerine adhezyonda biyotipler arasında istatistiksel olarak fark belirlendi. Sık izole edilen biyotiplerin adherens indekslerinin de yüksek olduğu görüldü.

Abstract:
The main purpose of this study was to determine immunodominant structural proteins of G. vaginalis biyotypes isolated from vaginal samples of women. The structural proteins of eight biotypes were first analysed by SDS-polyacrylamide gel electrophoresis, and subsequently blotted onto nitrocelulose membranes by electrophoresis. The antigenic proteins were then identified by reacting each of biotype proteins with homolog antiserum obtained by subsequent live bacteia injections into rabbits. In addition, the adhesion of the biotypes to the vaginal epithelial cells were studied with a view to identify whether there was any correlation between biotypes isolated more frequently from bacterial vaginosis cases and those that are more antijenic and serologically similar biotypes. SDS-PAGE analysis identified to many strongly expressed proteins, but no major differences were found between the number and/or the density of individual structural proteins as determined by coomassie blue stained gels. On the other hand, the immonoblotting studies indicated that among all the major structural proteins of G. vaginalis a protein band with an apparent molecular weight of 41 kDa was the most reactive with homolog as well as heterolog antisera. Although, this immunodominant protein was not readily identified in coomassie blue stained gels, it is strongly detected by the biotype spesific antiserum, and found to be cross reactive, albeit with weak reactions with some of the biotype antisera. Furthermore, the antigenic properties of the Biotypes 1, 2 and 5 were found to be strong and equally cross reactive while the biotypes 4, 7 and 8 were much less antigenic than the other biotypes. It was also shown that there was statistically significant differences between the biotypes in terms of adherence to the vaginal epithelial cells, and that those biotypes isolated more frequently in the bacterial vaginosis cases had higher adherence indexes.