Detection of cellular immunity to rabies antigens in human vaccinees.
A nonradioactive multi-parameter flow cytometry assay was developed to identify antigen-specific lymphocytes in human subjects previously vaccinated against rabies virus and was subsequently compared to the standard tritiated thymidine method. A cell tracking dye, carboxyfluorescein succinimidyl ester, was used in combination with surface label for CD4 and CD8 cells in order to determine the response of lymphocytes to killed rabies virus in an antigen recall assay. The rabies virus-specific lymphocyte response was compared to the humoral immune response in each of ten vaccinated and five non-vaccinated subjects. Lymphocyte responses to rabies virus were observed in all ten vaccinated subjects; some noted as early as 3 days after stimulation while others were not until 7 days after stimulation. There was good agreement between the proliferation index of the CFSE assay and the simulation index of the [3H]thymidine assay (kappa statistic=0.73). An inverse relationship was detected between the level of rabies virus neutralizing antibody (RVNA) and the lymphocyte response to inactivated rabies virus in the vaccinated subjects. The association between cytokines production and level of humoral and cellular response was investigated in four representative subjects. Two vaccinated subjects with high proliferation indices and low RVNA titers produced Th1 type cytokines to rabies virus stimulation, whereas two vaccinated individuals with low proliferation indices and high RVNA titers responses did not produce these cytokines.
Moore
Susan M
SM
Department of Diagnostic Medicine, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas 66506, USA. smoore@vet.ksu.edu
Wilkerson
Melinda J
MJ
Davis
Rolan D
RD
Wyatt
Carol R
CR
Briggs
Deborah J
DJ
D000328:Adult; D000914:Antibodies, Viral / Q000276:immunology*; D000917:Antibody Formation; D015704:CD4 Antigens / Q000276:immunology; D016827:CD8 Antigens / Q000276:immunology; D002460:Cell Line; D049109:Cell Proliferation; D016207:Cytokines / Q000096:biosynthesis; D005260:Female; D005434:Flow Cytometry; D005452:Fluoresceins; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D006801:Humans; D007111:Immunity, Cellular; D008297:Male; D008875:Middle Aged; D011819:Rabies Vaccines / Q000276:immunology*; D011820:Rabies virus / Q000276:immunology*; D013388:Succinimides; D014611:Vaccination; D014764:Viral Proteins / Q000276:immunology*
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D000914:Antibodies, Viral; D015704:CD4 Antigens; D016827:CD8 Antigens; D016207:Cytokines; D005452:Fluoresceins; D005456:Fluorescent Dyes; D011819:Rabies Vaccines; D013388:Succinimides; D014764:Viral Proteins