Concurrent production of interleukin-2, interleukin-10, and gamma interferon in the regional lymph nodes of mice with influenza pneumonia.
Cytokine production has been assessed at the single-cell level (ELISPOT assay) for freshly isolated mediastinal lymph node cells from C57BL/6 mice with primary, nonfatal influenza pneumonia. The mediastinal lymph node populations were also secondarily stimulated in vitro, and culture supernatants were assayed by enzyme-linked immunosorbent assay. Both approaches showed minimal evidence of protein secretion for interleukin-4 (IL-4), IL-5, and tumor necrosis factor, while IL-2, IL-10, and gamma interferon (IFN-gamma) were prominent throughout the response. The numbers of IL-2- and IFN-gamma-producing cells were maximal at 7 days after infection, while the total counts for cells secreting IL-10 were fairly constant from day 3 to 7. The cultures that were stimulated with virus in vitro showed in inverse relationship between IL-10 and IFN-gamma production, with IL-10 peaking on day 3 and IFN-gamma peaking on day 7. Lymphocytes secreting IL-2, IL-10, and/or IFN-gamma were present in CD4+ and CD8+ populations separated by fluorescence-activated cell sorting, although the CD8+ T cells produced less cytokine and were at a relatively lower frequency. Addition of recombinant IL-10 to the virus-stimulated cultures decreased the amount of IFN-gamma that could be detected, while incorporation of a monoclonal antibody to IL-10 had the opposite effect. A neutralization experiment also indicated that IL-2 was the principal mediator of lymphocyte proliferation. These experiments thus show that the developing T-cell response in the regional lymph nodes of mice with influenza cannot be rigidly categorized on the basis of a TH1 or TH2 phenotype and suggest possible regulatory mechanisms.
Sarawar
S R
SR
Department of Immunology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105.
Doherty
P C
PC
D000818:Animals; D016207:Cytokines / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D009980:Influenza A virus; D007371:Interferon-gamma / Q000096:biosynthesis*; D016753:Interleukin-10 / Q000096:biosynthesis; D007376:Interleukin-2 / Q000096:biosynthesis; D007378:Interleukins / Q000096:biosynthesis*; D008198:Lymph Nodes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008213:Lymphocyte Activation; D008482:Mediastinum / Q000033:anatomy & histology / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D009500:Neutralization Tests; D009976:Orthomyxoviridae Infections / Q000378:metabolism*; D010641:Phenotype; D011024:Pneumonia, Viral / Q000378:metabolism*; D013047:Specific Pathogen-Free Organisms; D016176:T-Lymphocyte Subsets / Q000378:metabolism
D016207:Cytokines; D007376:Interleukin-2; D007378:Interleukins; D016753:Interleukin-10; D007371:Interferon-gamma