Production of interleukin 3 and gamma-interferon by an antigen-specific mouse suppressor T cell clone.
An interleukin 2 (IL 2)-dependent, keyhole limpet hemocyanin (KLH)-specific, mouse suppressor T cell clone, 3D10, was found to produce interleukin 3 (IL 3) and gamma-interferon (IFN-gamma) in response to T cell mitogens Con A and PHA. Different from KLH-specific suppressor factor (TsF) that was spontaneously released into the medium when cultured in IL 2-containing conditioned medium, the production of IL 3 and IFN-gamma was induced by mitogenic stimuli. IL 3, IFN-gamma, and TsF were separable by gel filtration through a Sephadex G-100 column, being recovered in fractions of m.w. 25,000 to 30,000, 45,000 to 50,000 and 60,000 to 70,000, respectively. On the other hand, minimum size of IL 3 and IFN-gamma were shown to be about 25,000 and 20,000, respectively, by determining the lymphokine activities contained in the extracts from slices of SDS gels. These results indicate that IFN-gamma was present as a homodimer or hetero-complex with another carrier protein(s), whereas IL 3 was present as a monomeric form. A highly positive correlation (a correlation coefficient r = 0.96) between the titers of IL 3 and IFN-gamma produced by seven subclones derived from 3D10 was obtained, suggesting that IL 3 and IFN-gamma are induced by a process with a common mechanism. 3D10 also produced IL 3 and IFN-gamma when cultured with its specific antigen, KLH, in the presence of antigen-presenting cells. When Con A-stimulated 3D10 cells were labeled with L-[35S]methionine, we found that at least three proteins, with m.w. of 35,000, 25,000, and 20,000, were specifically released into medium by the stimulation. The latter two may be IL 3 and IFN-gamma described above, respectively, because of the similarities in m.w.
Koyasu
S
S
Nakauchi
H
H
Kitamura
K
K
Yonehara
S
S
Okumura
K
K
Tada
T
T
Yahara
I
I
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D002850:Chromatography, Gel; D002999:Clone Cells / Q000276:immunology / Q000378:metabolism; D003208:Concanavalin A / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D005260:Female; D000949:Histocompatibility Antigens Class II; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000302:isolation & purification; D007377:Interleukin-3; D008222:Lymphokines / Q000096:biosynthesis* / Q000302:isolation & purification; D008407:Mast Cells / Q000276:immunology; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D008811:Mice, Inbred DBA; D008819:Mice, Nude; D010641:Phenotype; D010835:Phytohemagglutinins / Q000494:pharmacology; D051381:Rats; D011917:Rats, Inbred Lew; D013491:Suppressor Factors, Immunologic; D050378:T-Lymphocytes, Regulatory / Q000145:classification / Q000276:immunology / Q000378:metabolism*
D000939:Epitopes; D000949:Histocompatibility Antigens Class II; D007377:Interleukin-3; D008222:Lymphokines; D010835:Phytohemagglutinins; D013491:Suppressor Factors, Immunologic; D003208:Concanavalin A; D007371:Interferon-gamma; D008715:Methionine