Phenotypic characterization of human CD4+ regulatory T cells obtained from cutaneous dinitrochlorobenzene-induced delayed type hypersensitivity reactions.
In this study, we describe the generation and characterization of cloned human CD4+ T lymphocyte populations that have infiltrated into cutaneous, 2,4-dinitrochlorobenzene-induced delayed type hypersensitivity reactions in healthy human subjects. It is shown that, in addition to T helper type 1 clones, elevated numbers of regulatory T clones, producing high levels of interleukin-10 and interleukin-5, but no measurable interleukin-4, were isolated from delayed type hypersensitivity reactions in four of six donors. A subsequent challenge with 2,4-dinitrochlorobenzene of two donors from whom only few interleukin-10-producing T cell clones had been generated after primary challenge, resulted in a decrease in the frequency of T helper type 1 clones and a strong increase in the number of interleukin-10-producing T helper type 2 and regulatory T clones. Culture supernatants from the latter cells, activated with anti-CD3 and anti-CD28 monoclonal antibody, inhibited alloantigen-mediated T cell proliferation which was, partly dependent on interleukin-10, and independent of transforming growth factor-beta. In addition, dendritic cells generated in vitro in the presence of these culture supernatants were impaired in their ability to induce alloantigen-induced proliferative responses. Differential expression of transcripts for the T1/ST2 molecule enabled a phenotypic distinction between resting regulatory T cells and T helper type 2 cells, but not between regulatory T cells and T helper type 1 cells. This experimental model provides a useful tool to isolate human inflammatory and anti-inflammatory T cell subpopulations and, furthermore, enables the study of the kinetics of their appearance into delayed type hypersensitivity reactions.
Lécart
S
S
INSERM U454, CHU Arnaud de Villeneuve, Montpellier, France.
Boulay
V
V
Raison-Peyron
N
N
Bousquet
J
J
Meunier
L
L
Yssel
H
H
Pène
J
J
D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology; D002478:Cells, Cultured; D017931:DNA Primers; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology; D004137:Dinitrochlorobenzene / Q000008:administration & dosage*; D015870:Gene Expression / Q000276:immunology; D006241:Haptens / Q000008:administration & dosage; D006801:Humans; D006968:Hypersensitivity, Delayed / Q000139:chemically induced / Q000276:immunology*; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000235:genetics / Q000276:immunology; D000072179:Interleukin-1 Receptor-Like 1 Protein; D016753:Interleukin-10 / Q000235:genetics; D015847:Interleukin-4 / Q000235:genetics / Q000276:immunology; D007509:Irritants / Q000008:administration & dosage*; D007519:Isoantigens / Q000235:genetics / Q000276:immunology; D008565:Membrane Proteins; D009000:Monocytes / Q000166:cytology / Q000276:immunology; D011506:Proteins / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D011956:Receptors, Cell Surface; D018123:Receptors, Interleukin / Q000235:genetics / Q000276:immunology; D053707:Receptors, Interleukin-12; D018418:Th2 Cells / Q000166:cytology / Q000276:immunology
D017931:DNA Primers; D004137:Dinitrochlorobenzene; D006241:Haptens; C079241:IL1RL1 protein, human; D000072179:Interleukin-1 Receptor-Like 1 Protein; D007509:Irritants; D007519:Isoantigens; D008565:Membrane Proteins; D011506:Proteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D018123:Receptors, Interleukin; D053707:Receptors, Interleukin-12; D016753:Interleukin-10; D015847:Interleukin-4; D007371:Interferon-gamma