Detection of Cellular Immunity to Rabies Antigens in Human Vaccinees
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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.
KEY WORDSCarboxyfluoroscein succinimidyl ester in vitro proliferation flow cytometry rabies vaccine response cytokines
We would like to thank the rabies vaccinated and non-vaccinated individuals who participated in this study. We are indebted to Tammy Koopman and Mehrdad Ameri for their technical assistance and review of the manuscript and to Teri Ricke for her help with the RFFIT testing.
- 4.Kuwert EK, Barsenbach J, Werner J, Marcus I, Scheiermann N, Grosse-Wilde H et al.: Early/high and late/low responders among HDCS vaccinees. In Cell Culture Rabies Vaccines and Their Protective Effect in Man, EK Kuwert, RJ Wiktor, H Koprowski (eds). Geneva, International Green Cross, 1981, pp 160–168Google Scholar
- 9.Lafon M: Immunology. In Rabies, AC Jackson, WH Wunner (eds). San Diego, CA, Academic Press, 2002, pp 351–371Google Scholar
- 16.Habel K: Habel test for potency. In Laboratory Techniques in Rabies, FX Meslin, MM Kaplan, H Koprowski (eds). Geneva, World Health Organization, 1996, pp 369-373Google Scholar
- 17.Centers for Disease Control and Prevention: Recommendations of the Advisory Committee on Immunization Practices (ACIP). 48: 1–21. United States, Morbidity and Mortality Weekly Report, 1999Google Scholar
- 18.World Health Organization: World Health Organization Expert Committee on Rabies: Eighth Report. 824. Geneva, WHO Press, WHO Technical Report Series, 1992Google Scholar
- 19.Altman DG: Practical Statistics for Medical Research. London, Chapman and Hall, 1991Google Scholar
- 22.Wells AD, Gudmundsdottir H, Turka LA: Following the fate of individual T cells throughout activation and clonal expansion. Signals from T cell receptor and CD28 differentially regulate the induction and duration of a proliferative response. J Clin Invest 100:3173–3183, 1997PubMedCrossRefGoogle Scholar