Induction of interleukin-6 in murine bone marrow-derived macrophages stimulated by the Mycoplasma arthritidis mitogen MAS.
Cell-free supernatant of cultures from Mycoplasma arthritidis (MAS) functions as an extremely potent T-cell mitogen for human and murine lymphocytes. The T-cell response is dependent on the presence of accessory cells, presenting the intact E2 molecule on the cell surface. Until now, pure MAS protein has not been available. We developed a new multi-step method for MAS purification. The main steps in this protocol are ammonium sulfate precipitation, anion exchange and hydroxyapatite chromatography followed by gel filtration. With this efficient protocol we obtained fractions of extremely potent mitogenic properties, the purification rate was about 5 x 10(5). Although this protease-sensitive mitogenic activity was highly enriched, we failed to detect the protein by sensitive staining methods of SDS-PAGE. In previous studies, we showed that MAS induces the synthesis of interferon gamma in human and murine lymphocyte cultures. Here we demonstrate that MAS induces interleukin-6 (IL-6) in murine bone-marrow derived macrophage cultures. Since IL-6 is also induced by endotoxin, we used C3H/HeJ mice, which are known to be LPS-nonresponders, in all our studies.
Homfeld
J
J
Institute of Virus Research, German Cancer Research Center, Heidelberg.
Homfeld
A
A
Nicklas
W
W
Rink
L
L
Weyland
A
A
Kirchner
H
H
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D000941:Antigens; D000942:Antigens, Bacterial; D001853:Bone Marrow / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000276:immunology; D008264:Macrophages / Q000187:drug effects*; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008934:Mitogens / Q000302:isolation & purification / Q000494:pharmacology*; D011506:Proteins; D012313:RNA / Q000032:analysis / Q000302:isolation & purification; D018089:Superantigens; D013601:T-Lymphocytes / Q000276:immunology*
D000941:Antigens; D000942:Antigens, Bacterial; D015850:Interleukin-6; D008934:Mitogens; C052996:Mycoplasma arthritidis mitogen; D011506:Proteins; D018089:Superantigens; D012313:RNA