Purification of a factor which provides a costimulatory signal for gamma interferon production.
A protein factor which induces high levels of gamma interferon (IFN-gamma) in resting splenic nonadherent cells was isolated from the sera of mice with generalized inflammation caused by endotoxic shock. The factor was highly purified by ammonium sulfate precipitation followed by ion-exchange column chromatography on DEAE-Sepharose, molecular sieving on Ultrogel AcA 44, and hydrophobic column chromatography with phenyl-Sepharose. It was further purified to apparent homogeneity by polyacrylamide gel electrophoresis. It induced IFN-gamma production in a dose-dependent manner in the presence of interleukin-2, monoclonal anti-CD3 antibody (anti-CD3 MAb), or concanavalin A (ConA) in spleen cells deprived of plastic plate- and nylon wool-adherent cells. Anti-CD3 MAb induced the highest level of production of the three. The factor, interleukin-2, anti-CD3 MAb, or ConA alone induced a trace of or no detectable IFN-gamma in these cells. The factor also exhibited an accessory function during proliferation in these cells in the presence of a suboptimal dose of ConA. However, the factor failed to stimulate IFN-gamma production when staphylococcal enterotoxin A, a superantigenic T-cell mitogen, was employed. Treatment with pronase or heat abolished these activities. These studies confirm the existence of a soluble protein factor which is able to exhibit a novel accessory function in IFN-gamma production in resting T or natural killer cells. It will be of interest to compare this factor with the recently cloned human natural killer stimulatory factor (NKSF/IL-12).
Nakamura
K
K
Department of Bacteriology, Hyogo College of Medicine, Japan.
Okamura
H
H
Nagata
K
K
Komatsu
T
T
Tamura
T
T
D000818:Animals; D017252:CD3 Complex / Q000502:physiology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003208:Concanavalin A / Q000494:pharmacology; D004306:Dose-Response Relationship, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D004768:Enterotoxins / Q000494:pharmacology; D006358:Hot Temperature / Q000009:adverse effects; D007371:Interferon-gamma / Q000096:biosynthesis*; D018664:Interleukin-12; D007376:Interleukin-2 / Q000494:pharmacology; D007378:Interleukins / Q000276:immunology; D007694:Killer Cells, Natural / Q000276:immunology; D051379:Mice; D008809:Mice, Inbred C3H; D011402:Pronase / Q000494:pharmacology; D013601:T-Lymphocytes / Q000276:immunology
D017252:CD3 Complex; D004768:Enterotoxins; D007376:Interleukin-2; D007378:Interleukins; D003208:Concanavalin A; D018664:Interleukin-12; C026066:enterotoxin A, Staphylococcal; D007371:Interferon-gamma; D011402:Pronase