Antigen-dependent cytokine mRNA expression by individual rhesus macaque T helper cells by flow cytometry.
Specific patterns of cytokine secretion by CD4(+) T helper (Th) cells determine the nature of immune effector responses. Using a multiparameter, flow cytometric fluorescent in situ hybridization (FISH) assay that detected cytoplasmic mRNA within intact cells, we assessed antigen-specific cytokine expression in rhesus macaque Th cells. In the peripheral lymphocytes of immunized rhesus macaques, FISH detected antigen-induced cytokine gene expression in single Th cells. Analysis of simultaneous cytokine expression by single cells demonstrated that the recall immune response consisted of Th cells expressing either a Th1 (IL-2(+)/IFN-gamma(+)) or a Th2 (IL-4(+)/IL-6(+)) cytokine pattern. In addition to the classic Th subsets, Th cells expressing only one of two Th1 or Th2 defining cytokines were common following antigen restimulation. The data gathered with the FISH assay suggest that, in primates, the immune response to recall antigens consists of nonclassic Th cells, as well as a mixture of polarized Th1 and Th2 T cells.
Arredondo
J
J
California Regional Primate Research Center, School of Veterinary Medicine, Davis 95616, USA.
Xusheng Lü
F
F
Villinger
F
F
McChesney
M B
MB
McGhee
J R
JR
Miller
C J
CJ
D000818:Animals; D000941:Antigens / Q000276:immunology; D016207:Cytokines / Q000235:genetics / Q000302:isolation & purification*; D005434:Flow Cytometry / Q000379:methods; D006433:Hemocyanins / Q000276:immunology; D007114:Immunization; D017404:In Situ Hybridization, Fluorescence; D007371:Interferon-gamma / Q000235:genetics / Q000302:isolation & purification; D007376:Interleukin-2 / Q000235:genetics / Q000302:isolation & purification; D015847:Interleukin-4 / Q000235:genetics / Q000302:isolation & purification; D015850:Interleukin-6 / Q000235:genetics / Q000302:isolation & purification; D008253:Macaca mulatta; D012333:RNA, Messenger / Q000302:isolation & purification*; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology
D000941:Antigens; D016207:Cytokines; D007376:Interleukin-2; D015850:Interleukin-6; D012333:RNA, Messenger; D015847:Interleukin-4; D007371:Interferon-gamma; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin