caa a22 4500 469073543 CHVBK 20180323132928.0 cr unu---uuuuu 170328e19920101xx s 000 0 eng 10.1007/BF01741048 doi (NATIONALLICENCE)springer-10.1007/BF01741048 Ryoyama Kazuo Department of Experimental Therapeutics, Cancer Research Institute, Kanazawa University, 13-1 Takara-machi, 920, Kanazawa, Japan aut Effector molecules from antitumor macrophages induced with OK-432 and cyclophosphamide [Elektronische Daten] [Kazuo Ryoyama] Summary: The present study was designed to determine whether antitumor activity of macrophages induced with OK-432 and cyclophosphamide was mainly dependent on their ability to produce a soluble factor, that is,l-arginine-dependent nitric oxide as measured by nitrite concentration. Nitrite production by peritoneal macrophages from NIH Swiss mice pretreated with OK-432 (125 KE/kg) i.p. twice at 1-week intervals and with cyclophosphamide (200 mg/kg) i.p. 2 days before the second OK-432 treatment, increased with time for 24 h, and proportionally depended on macrophage numbers. Nitrite production was inhibited by actinomycin D and puromycin but not by mitomycin C.N G-Monomethyl-l-arginine, a specific competitive inhibitor ofl-arginine-dependent nitric oxide synthesis, also inhibited production. There was a close correlation between nitrite production and antitumor activity in macrophages from mice pretreated with either OK-432 and cyclophosphamide, OK-432, or thioglycolate broth. OK-432 increased both nitrite production and antitumor activities when added to the macrophage from mice pretreated with OK-432 but not with thioglycolate broth. Both activities of macrophages from mice pretreated with OK-432 and cyclophosphamide were enhanced with increasing concentrations ofl-arginine (0.125-1 mM) in the culture medium.d-Arginine, however, did not substitute forl-arginine. Neither activity was affected by contact between the macrophage and the EL4 cell. The macrophage showed antitumor activity through a membrane filter though the activity was greatly reduced. This antitumor activity of macrophages through a membrane was also inhibited byN G-Monomethyl-l-arginine, and increased by OK-432. However, conditioned media, obtained by culturing macrophages induced with OK-432 and cyclophosphamide, inhibited growth of EL4 cells. This activity was carried out by dialysable and non-dialysable factors. One of the dialysable factors was nitrite, an oxidized product of nitric oxide. The antitumor activity of non-dialysable factors was heat-stable and production of factors was increased byN G-Monomethyl-l-arginine and OK-432. Also, non-dialysable factors increased both antitumor and nitrite production activities of OK-432-elicited macrophages, when incubated with factors. Such activity of factors was also heat-stable. The production of factors increased with incubation time of macrophages, and was not inhibited byN G-Monomethyl-l-arginine. These results indicate that in vitro antitumor activity of macrophages induced with OK-432 and cyclophosphamide was mainly dependent onl-arginine-dependent nitric oxide, and that macrophageassociated soluble factors other than nitric oxide were also needed to inhibit fully tumor growth in vitro. Springer-Verlag, 1992 Macrophages nationallicence Nitric oxide nationallicence TNF nationallicence OK-432 nationallicence Cyclophosphamide nationallicence Cancer Immunology, Immunotherapy Springer-Verlag 35/1(1992-01-01), 7-13 0340-7004 35:1<7 1992 35 262 https://doi.org/10.1007/BF01741048 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01741048 text/html Onlinezugriff via DOI NATIONALLICENCE 100 1- Ryoyama Kazuo Department of Experimental Therapeutics, Cancer Research Institute, Kanazawa University, 13-1 Takara-machi, 920, Kanazawa, Japan aut NATIONALLICENCE 773 0- Cancer Immunology, Immunotherapy Springer-Verlag 35/1(1992-01-01), 7-13 0340-7004 35:1<7 1992 35 262 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer