Production of hapten-specific T cell hybridomas and their use to study the effect of ultraviolet B irradiation on the development of contact hypersensitivity.
We produced a series of T cell hybridomas that produce IL-2 when cultured with syngeneic APC coupled to FITC or TNP. These hybridomas are hapten specific and Ia restricted. The hybridomas were used to detect hapten-bearing APC in draining lymph nodes of mice sensitized with trinitrochlorobenzene or FITC in vivo. Hapten-bearing APC capable of stimulating the hybridomas were detectable in draining lymph nodes of hapten-painted mice within 3 h after sensitization. The ability of lymph node APC to stimulate the hybridomas peaked at 24 h and declined by 48 h. The dendritic cell subpopulation was the subpopulation of cells that were found in the regional lymph nodes of hapten-painted animals that were capable of stimulating the hybridomas to produce IL-2. Prior treatment of the skin with low dose UVB irradiation before epicutaneous application of contact sensitizers significantly reduced the capacity of hapten-bearing APC to stimulate the hybridomas. This observation was corroborated by results obtained from flow microfluorometry analysis of lymph node cells from FITC-sensitized mice. Lymph node dendritic cells obtained from FITC-painted mice contain a brightly staining group of cells by flow microfluorometry analysis. Lymph node dendritic cells from FITC-painted, UVB-irradiated mice did not contain this brightly staining population. These results indicate that low dose, local UVB irradiation may affect APC migration and/or function. We believe that these hybridomas will prove to be useful tools in the study of the development and regulation of contact hypersensitivity.
Bigby
M
M
Department of Pathology, Massachusetts General Hospital, Boston.
Vargas
R
R
Sy
M S
MS
D000818:Animals; D000938:Antigen-Presenting Cells / Q000032:analysis; D003877:Dermatitis, Contact / Q000209:etiology / Q000276:immunology*; D004817:Epidermis / Q000528:radiation effects; D000939:Epitopes / Q000276:immunology*; D005260:Female; D006241:Haptens / Q000276:immunology*; D006825:Hybridomas / Q000032:analysis / Q000276:immunology / Q000528:radiation effects*; D007801:Langerhans Cells / Q000528:radiation effects; D008198:Lymph Nodes; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D013601:T-Lymphocytes / Q000276:immunology / Q000528:radiation effects*; D014301:Trinitrobenzenes / Q000276:immunology; D014466:Ultraviolet Rays
D000939:Epitopes; D006241:Haptens; D014301:Trinitrobenzenes