title: Flow cytometric assessment of human peripheral blood mononuclear cells in response to a coccidioidal antigen.
abstract: The in vitro responses of peripheral blood mononuclear cells (PBMC) from healthy immune and non-immune donors were assessed by flow cytometry after incubation with the coccidioidal antigen toluene spherule lysate (TSL). After 120 h of incubation with 100 microg ml(-1) of TSL, expression of the activation markers CD69, CD25 and human leukocyte antigen-DR were all significantly increased in CD3+ lymphocytes from immune donors compared to non-immune donors (P < 0.03 for all). No differences in the surface expression of the costimulatory molecules CD28, CD152 or CD154 was seen between immune and non-immune donors after either 24 or 120 h of TSL incubation, nor were differences detected in the expression of the B7 ligands CD80 or CD86 on CD14+ monocytes. The percent of CD3+ lymphocytes expressing intracellular interferon-gamma (IFN-gamma) was significantly increased in immune compared to non-immune donors and was further increased by the addition of 10 ng ml(-1) of human recombinant interleukin (IL)-12 (P < 0.05 for both). Both CD4+ and CD8+ lymphocytes contributed to IFN-gamma production. These data indicate that coccidioidal antigen stimulation of lymphocytes from healthy immune donors leads to specific expression of activation molecules and production of intracellular IFN-gamma. Addition of IL-12 leads to a significant recruitment of cells producing IFN-gamma among immune donors.
authors.affiliation: University of Arizona and the Medical and Research Services of the Tucson Veterans Affairs Medical Center, 85723, USA. nampel@u.arizona.edu
authors.forename: N M
authors.initials: NM
authors.lastname: Ampel
authors.forename: L
authors.initials: L
authors.lastname: Christian
mesh_terms: D000069594:Abatacept; D015703:Antigens, CD / Q000032:analysis; D000943:Antigens, Differentiation / Q000032:analysis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D000946:Antigens, Fungal / Q000494:pharmacology*; D018106:CD28 Antigens / Q000032:analysis; D017252:CD3 Complex / Q000032:analysis; D023201:CD40 Ligand; D060908:CTLA-4 Antigen; D002478:Cells, Cultured; D003045:Coccidioides / Q000276:immunology*; D003047:Coccidioidomycosis / Q000097:blood / Q000276:immunology / Q000382:microbiology; D016207:Cytokines / Q000032:analysis; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D007111:Immunity, Cellular; D018796:Immunoconjugates; D007371:Interferon-gamma / Q000032:analysis; D018664:Interleukin-12 / Q000494:pharmacology; D037181:Lectins, C-Type; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000032:analysis; D015375:Receptors, Interleukin-2 / Q000032:analysis; D013997:Time Factors; D000099192:CD69 Antigens
chemical_list: D000069594:Abatacept; D015703:Antigens, CD; D000943:Antigens, Differentiation; D000945:Antigens, Differentiation, T-Lymphocyte; D000946:Antigens, Fungal; D018106:CD28 Antigens; D017252:CD3 Complex; D023201:CD40 Ligand; D060908:CTLA-4 Antigen; D016207:Cytokines; D006684:HLA-DR Antigens; D018796:Immunoconjugates; D007371:Interferon-gamma; D018664:Interleukin-12; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D015375:Receptors, Interleukin-2; D000099192:CD69 Antigens; C556706:CTLA4 protein, human