Interleukin-3 and encephalitogenic activity of SJL/J myelin basic protein-specific T cell lines.
Myelin basic protein (MBP)-specific SJL/J T cells were cultured in normal growth medium or growth medium supplemented with 10% culture supernatant from WEHI-3 cells, a source of interleukin-3 (IL-3), or with recombinant IL-3. T cell lines cultured with IL-3 supplementation were more encephalitogenic compared to parallel lines cultured without this supplement. There was little difference in antigen-specific proliferative response or expression of cell surface markers CD3, CD4, CD8, IL-2R, or alpha/beta TCR in the parallel lines. Supernatant fluids from antigen-stimulated T cells from each cycle were tested for the presence of IL-2, IL-3, IL-4, granulocyte macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF alpha/beta) and transforming growth factor-beta (TGF beta). No significant difference in IL-2, IL-4, GM-CSF, TNF alpha/beta, or TGF beta levels were seen when supplemented and unsupplemented cultures were compared. Supernatant culture fluids contained an activity that was highly stimulatory for the IL-3-dependent mouse mast cell line, MC/9. This activity was attributable to a combination of at least three factors that varied in relative concentrations throughout the course of the experiments. Based on neutralization by monoclonal antibodies, MC/9 stimulating activity in early passage lines was attributable entirely to IL-3 and GM-CSF. The fraction of the MC/9 stimulatory activity that could be neutralized by monoclonal antibody to IL-3 decreased with increasing stimulation cycle while the fraction neutralized by anti-GM-CSF antibodies remained relatively constant. At the time that the lines lost encephalitogenicity, the activity neutralizable by anti-IL-3 had dropped to low levels in the culture supernatants; however, MC/9 stimulatory activity remained present in the supernatants. This was due to GM-CSF and a third unidentified factor.
Zhao
M L
ML
Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226.
Xia
J Q
JQ
Fritz
R B
RB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000209:etiology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000502:physiology; D007376:Interleukin-2 / Q000032:analysis / Q000502:physiology; D007377:Interleukin-3 / Q000502:physiology*; D051379:Mice; D004676:Myelin Basic Protein / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000502:physiology
D000911:Antibodies, Monoclonal; D007376:Interleukin-2; D007377:Interleukin-3; D004676:Myelin Basic Protein; D014409:Tumor Necrosis Factor-alpha; D016178:Granulocyte-Macrophage Colony-Stimulating Factor