Generation of functional murine macrophage lines employing a helper-free and replication-defective SV40-retrovirus: cytokine-dependent growth.
By using a helper-free and replication-defective recombinant retrovirus encoding the SV40 early antigens (MV40), we have established continuous macrophage (M phi) lines. All of the lines were nonproducer M phi's with differentiated M phi functions such as phagocytosis, cytotoxicity, and IL-1 and TNF production. To determine the effects of several cytokines on growth of mature M phi's, the responsiveness of these established M phi lines to various cytokines was investigated in methylcellulose culture. Their response patterns to several cytokines alone and in combination were different, implying that there might be mature M phi subpopulations with distinct growth profiles regulated by several cytokines. On the other hand, all of the lines efficiently yielded a number of colonies in response to interleukin-4 (IL-4) alone. Moreover, IL-4 cooperated with interleukin-3 (IL-3) to enhance colony formation of all the lines. A similarly synergistic effect was observed in combination of IL-4 and macrophage-colony stimulating factor (M-CSF) in almost all the lines. Similar results were obtained with colony formation of fresh thioglycolate-induced M phi's. These observations suggested that IL-4 was involved in growth of mature M phi's. Our present results suggest that the helper-free and replication-defective MV40 is of use to obtain continuous and functional cell lines from primary M phi's.
Ogawa
Y
Y
Biomaterial Research Institute Co., Ltd., Kanagawa, Japan.
Ohno
N
N
Ito
M
M
Iizuka
M
M
Kobayashi
S
S
Sudo
T
T
D000818:Animals; D000957:Antigens, Viral, Tumor / Q000235:genetics; D015139:Blotting, Southern; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology; D016207:Cytokines / Q000502:physiology*; D003601:Cytotoxicity Tests, Immunologic; D006378:Helper Viruses / Q000502:physiology*; D008264:Macrophages / Q000166:cytology*; D051379:Mice; D011233:Precipitin Tests; D012190:Retroviridae / Q000472:pathogenicity*; D013539:Simian virus 40 / Q000472:pathogenicity*; D014412:Tumor Virus Infections / Q000503:physiopathology*; D016662:Virus Integration / Q000502:physiology; D014779:Virus Replication
D000957:Antigens, Viral, Tumor; D016207:Cytokines