Quantitative assessment by flow cytometry of T-lymphocytes producing antigen-specific gamma-interferon in Brucella immune cattle.
Peripheral blood mononuclear cells (PBMC) isolated from cattle infected with Brucella secreted gamma-interferon (IFN-gamma) after antigen-specific stimulation with Brucellergene, which is a mixture of cytoplasmic proteins of rough Brucella melitensis B115. Following the depletion of the monocyte-macrophages from the PBMC, the enriched lymphocyte populations stimulated with Brucellergene did not produce IFN-gamma. Two-colour immunofluorescence staining of intracellular IFN-gamma and bovine cell surface molecules identified the cells producing IFN-gamma among the PBMC stimulated with Brucellergene. Moreover, this method could be used to estimate the number of T-cells specifically producing IFN-gamma. For a given animal, there is a significant correlation (p < 0.05) between the production of IFN measured by an ELISA of the supernatant of whole blood stimulated with Brucellergene and the number of T-cells producing IFN-gamma after in vitro stimulation with Brucellergene. The development of the immunofluorescence staining technique provides a new tool for analysing and for measuring the T-cell immune response in cattle.
Weynants
V
V
Laboratoire d'Immunologie-Microbiologie, U.R. Biologie Moléculaire, Facultés Universitaires Notre-Dame de la Paix, Namur, Belgium.
Walravens
K
K
Didembourg
C
C
Flanagan
P
P
Godfroid
J
J
Letesson
J J
JJ
D000818:Animals; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000276:immunology; D002002:Brucella / Q000276:immunology; D002004:Brucella Vaccine / Q000276:immunology*; D002003:Brucella abortus / Q000276:immunology; D002007:Brucellosis, Bovine / Q000276:immunology* / Q000517:prevention & control; D002417:Cattle; D002469:Cell Separation / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary; D005260:Female; D005434:Flow Cytometry / Q000379:methods / Q000662:veterinary; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*
D000942:Antigens, Bacterial; D002004:Brucella Vaccine; C005641:brucellin; D007371:Interferon-gamma