Clinical & Experimental Immunology · 1998 · 16 citations · 22 references
Life SciencesImmunologyMicrobiologyImmunology Of Infectious DiseasesMolecular VirologyVirologyAntiviral ImmunityCytokine BiologyImmune Responses To InfectionRespiratory InfectionsT Cell BiologyViral InfectionsAntiviral Cytokine ResponsesCellular Immune Response To VirusesCytokine MaturationInflammatory Response To Viral InfectionInflammatory Responses To Viral InfectionInterferon Gamma ProductionIntracellular ExpressionViral Infection Of Immune Cells
We examined changes in the cytokine profile of T cells induced by in vitro infection with RSV. Isolated mononuclear cells from 27 healthy adults and six infants were infected with RSV at a concentration of 3 MOI (multiplicity of infection). After 48 h cells were restimulated with phorbol ester and ionomycin in the presence of monensin for 5 h. The intracellular expression of viral antigen, the cytokines IL-2, IL-4, IL-5, interferon-gamma (IFN-gamma), and the expression of surface markers were assessed by immunofluorescent staining and flow cytometry. There was a significant (P<0.001) rise of IL-5 expression in RSV-infected cultures in comparison with uninfected cultures from the same individuals, whereas there were no changes in the expression of the other lymphokines. The increase in IL-5 generation depended on viable infectious RSV rather than inactivated virus. RSV infection as well as IL-5 production in T cells were confined to the CD8 subpopulation. However, there was no simultaneous expression of RSV antigen and IL-5. Purified T cells did not show any increase in IL-5 generation. However, when the rate of RSV infection was enhanced in monocytes by means of a specific monoclonal antibody, co-cultured T cells displayed an increase of IL-5 production compared with samples with ordinary low rate RSV infection. It is therefore likely that the increased commitment of lymphocytes to produce IL-5 after RSV infection in vitro is mediated by monocytes or other antigen-presenting cells.
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Detection of intracellular cytokines by flow cytometry
Thomas Jung, Christoph Heusser, Christine Neumann et al. · Journal of Immunological Methods · 1993 · 988 citations
R A Seder, Ricardo T. Gazzinelli, Ali Sher et al. · Proceedings of the National Academy of Sciences · 1993 · 981 citations