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Determination of cytokine co‐expression in individual splenic CD4 + and CD8 + T cells from influenza virus‐immune mice
Author(s) -
R. Falchetti,
Paolo Di Francesco,
Giulia Lanzilli,
Roberta Gaziano,
I. A. Casalinuovo,
Anna Teresa Palamara,
Giampietro Ravagnan,
Enrico Garaci
Publication year - 1998
Publication title -
immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.297
H-Index - 133
eISSN - 1365-2567
pISSN - 0019-2805
DOI - 10.1046/j.1365-2567.1998.00608.x
Subject(s) - biology , immune system , spleen , cd8 , cytokine , cytotoxic t cell , virus , interferon gamma , influenza a virus , microbiology and biotechnology , interleukin 4 , t cell , t lymphocyte , interleukin 21 , interleukin 2 , immunology , virology , in vitro , biochemistry
We have studied the patterns of interleukin‐2 (IL‐2), IL‐4 and interferon‐γ (IFN‐γ) co‐expression displayed by individual splenic CD4 + and CD8 + T cells in response to influenza virus immunization. Unseparated spleen cells obtained from mice intraperitoneally (i.p.) injected with A/PR8 (H1N1) influenza virus (PR8) were cultured for 24 hr in the presence of ultraviolet‐inactivated PR8. As controls, cultures of both naive spleen cells stimulated with PR8 or of immune cells lacking the inactivated virus were used. The frequencies of CD4 + and CD8 + T cells expressing IL‐2, IL‐4 and IFN‐γ were determined by three‐colour flow cytometric analysis of fixed and saponin‐permeabilized cells fluorescent‐stained for either CD4 or CD8 surface molecules and for one of the following combinations of two intracellular cytokines: IL‐2/IL‐4, IL‐2/IFN‐γ and IL‐4/IFN‐γ. The results showed that immunization with influenza virus induces in both CD4 + and CD8 + T cells a heterogeneity of cytokine response patterns that do not follow the type 1/type 2 polarized response model, but with substantial differences between the two populations. In fact, the analysis of the phenotypes of virus‐immune CD8 + T cells revealed similar significant proportions of cells either expressing any one of the three cytokines or co‐expressing combinations of them (i.e. IL‐4/IL‐2, IL‐4/IFN‐γ and IL‐2/IFN‐γ), whereas immune CD4 + T cells were seen to express almost exclusively a single cytokine per cell. The observed patterns of cytokine production suggest that influenza virus immunization induces the expression of a type 0 cytokine pattern at both population and single cell levels in CD8 + T cells and exclusively at the population level in CD4 + T cells.