Diversity among abortion strains of Chlamydia psittaci demonstrated by inclusion morphology, polypeptide profiles and monoclonal antibodies

Polymorphic outer-membrane proteins of Chlamydophila abortus are glycosylated

“…They can be separated on 20 i 20 cm gels, as shown previously (Vretou et al, 1996), and can be identified by two-dimensional electrophoresis (Giannikopoulou et al, 1997). In the present study we have established mAbs JA6C7 and EB3G2 as markers for two proteins, POMPs 90 and 91B, on the basis of antigenic profiles in one-and two-dimensional gels.…”

“…Further immunoblotting analysis with a panel of specific monoclonal antibodies (mAbs) revealed the presence of a triplet at 90 kDa and a doublet at 85 kDa, instead of a single protein (Souriau et al, 1994 ;Vretou et al, 1996). Consequently, the term ' antigen family at 90 kDa' was introduced ; it was suggested that the observed pleiomorphy might represent the differential trimming of a single protein or correspond to discrete products from different genes (Vretou et al, 1996). Screening of a λ gt11gDNA expression library with affinity-purified antibodies against the 90 kDa proteins identified two gene fragments (Longbottom et al, 1996).…”

“…Chlamydiae were grown in McCoy cells (ATCC, Manassas, VA, USA) and were monitored regularly for mycoplasma contamination with 4h,6-diamidine-2h-phenylindole dihydrochloride (DAPI, Boehringer Mannheim). Elementary bodies (EBs) were purified by centrifugation through a discontinuous gradient of gastrografin, as described previously (Vretou et al, 1996). Metabolic labelling of chlamydiae.…”

“…mAbs. The anti-POMP mAbs 181 and 8976\073, and mAb 4H9 against the POMP-like protein at 105 kDa, have been described previously, as has mAb 188 against the major outer-membrane protein (MOMP) (Vretou et al, 1996 ;Giannikopoulou et al, 1997). mAb ABF8, an IgG3, which recognizes EF-Tu (heat-labile elongation factor) in Western blots after two-dimensional electrophoresis, was produced in this laboratory.…”

Chlamydial abortion of ewes and goats. I. Current aspects on etiology, pathogenesis, immune mechanisms and epizootiology of disease

“…4 Ωστόσο, διάφορες μεταξύ των στελεχών έχουν βρεθεί όταν αυτά ελέγχθηκαν με διασταυρωμένες αντι δράσεις σε μοντέλο ποντικού. 3940 Οι διάφορες σε ορισμένα στελέχη επιβεβαιώθηκαν και in vitro με την εξέταση της μορφολογίας των εγκλείστων, την αντίδραση με μονο-κλωνικά αντισώματα, την ηλεκτροφόρηση των πρωτεϊνών τους 41 καθώς και με την εφαρμογή της PCR-RFLP στο δια-γονιδιακό τμήμα του 16S και 23S rRNA τους (προσωπική εμπειρία). Εξάλλου, με την εφαρμογή της PCR-AFLP, στελέχη της C abortus ομαδοποιήθηκαν ανάλογα με τη γε ωγραφική τους προέλευση και πιθανόν τον ξενιστή (πρό βατο, αίγα, βοοειδή) και τον ιστό απομόνωσης τους (απο βολής από εντερικά).…”