title: Characterization of the antigenic and functional domains of a Mycoplasma synoviae variant vlhA gene.
abstract: The Mycoplasma synoviae haemagglutinin gene, vlhA, encodes two major immunodominant and surface-exposed membrane proteins, MSPB and MSPA. Both products are antigenically variable but only MSPA mediates binding to erythrocytes. Previously we have shown that M. synoviae type strain WVU 1853 could express a variant vlhA gene, referred to as MS2/28.1, with a considerably shorter and divergent MSPA region. A finding that prompted detailed characterization of its antigenic and functional properties. Here we mutagenized each of the six opal codons of the variant MS2/28.1 vlhA member into tryptophan, thus allowing its expression in Escherichia coli as well as its cleavage products, MSPB and MSPA. In addition, we expressed 5 contiguous regions of MS2/28.1 extending from the last part of MSPB to the C-terminus of MSPA. Colony immunostaining with region-specific antisera mapped antigenic variation to the N-terminal half of MS2/28.1 MSPA. No haemagglutinating activity was observed for MSPB, but consistent haemadsorption inhibition was mapped to the region extending from amino acid 325 to 344. Inhibition of both haemagglutination and haemadsorption activities were obtained with sera directed against the C-terminal region of MSPA, with the highest titers (1/320 and 1/160, respectively) being recorded for its last 60 residues. Importantly, antibodies to this region also yielded the highest metabolic inhibition titer of 1/1280. Overall, aside from mapping the functional domains of a M. synoviae highly divergent haemagglutinin gene, this study shows that the C-terminal half of its MSPA region induced the highest titers of antibodies inhibiting haemagglutination, haemadsorption, and metabolism.
authors.affiliation: Laboratoire des Mycoplasmes, Institut Pasteur de Tunis, 13, Place Pasteur B.P. 74, 1002 Tunis-Belvédère, Tunis, Tunisia. awatef.bejaoui@yahoo.fr
authors.forename: Awatef Béjaoui
authors.initials: AB
authors.lastname: Khiari
authors.forename: Boutheina Ben Abdelmoumen
authors.initials: BB
authors.lastname: Mardassi
mesh_terms: D000818:Animals; D000940:Antigenic Variation; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D002645:Chickens / Q000276:immunology / Q000382:microbiology; D005260:Female; D006382:Hemadsorption; D006383:Hemadsorption Inhibition Tests; D006385:Hemagglutination Inhibition Tests; D006386:Hemagglutination Tests; D006388:Hemagglutinins / Q000235:genetics* / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D037102:Lectins / Q000235:genetics* / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D016297:Mutagenesis, Site-Directed; D045805:Mycoplasma synoviae / Q000235:genetics* / Q000276:immunology; D011817:Rabbits
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D006388:Hemagglutinins; D007106:Immune Sera; D037102:Lectins; D008565:Membrane Proteins; C406455:VlhA protein, Mycoplasma synoviae