Bovine IgG1 antibodies against Mycobacterium avium subsp. paratuberculosis protein p34-cx improve association of bacteria and macrophages.
Paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis (Map), is a chronic granulomatous enteric disease in cattle. Among molecular components of Map, protein p34 was identified as specific and immunodominant for bovine B cells. In order to determine if specific antibodies could influence the course of Map pathogenesis, the interaction between bacteria and bovine macrophages was studied. Bovine polyclonal antibodies from 3 calves vaccinated with protein p34-cx, 6 calves vaccinated with heat-killed Map, 8 naturally infected, and 3 healthy calves -as negative controls- were used. Specific anti-Map, -p34-cx and -PPA-3 antibodies were evaluated and isotype characterized. Infected and Map vaccinated animals showed similar IgG1 and IgG2 response against Map whole bacteria. When p34-cx was used as the antigen, mainly IgG1 and IgG3 were detected in infected and only IgG1 in p34-cx vaccinated animals. Bovine polyclonal antibodies from three animals of each category were isolated and affinity purified through Map and p34-cx columns. The effect of these antibodies in association with Map and a transformed bovine peritoneal macrophage's cell line (Bov-Mac) as well as activation of NF-kappaB transcription factor was studied. Our results show that association of Map significantly increased in vitro after pretreatment with bovine anti-Map or anti-p34-cx antibodies obtained from vaccinated or infected cattle when compared with those of controls. Improved activation of NF-kappaB was detected in macrophages that ingested Map opsonized with either anti-Map or anti-p34-cx specific antibodies of infected or vaccinated calves, suggesting that both anti-Map and IgG1 anti-p34-cx antibodies support Map-macrophage interactions.
Mundo
Silvia L
SL
Cátedra de Inmunología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Argentina. smundo@fvet.uba.ar
Fontanals
Adriana M
AM
García
Mariana
M
Durrieu
María
M
Alvarez
Elida
E
Gentilini
Elida R
ER
Hajos
Silvia E
SE
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology*; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000276:immunology*; D001428:Bacterial Vaccines / Q000276:immunology*; D002417:Cattle; D002418:Cattle Diseases / Q000097:blood / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D007132:Immunoglobulin Isotypes; D008264:Macrophages / Q000276:immunology* / Q000382:microbiology; D017737:Macrophages, Peritoneal / Q000276:immunology / Q000382:microbiology; D016927:Mycobacterium avium subsp. paratuberculosis / Q000276:immunology*; D016328:NF-kappa B / Q000378:metabolism; D010283:Paratuberculosis / Q000097:blood / Q000276:immunology
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D016328:NF-kappa B