Molecular mechanisms underlying IFN-gamma-mediated tumor growth inhibition induced during tumor immunotherapy with rIL-12.
The present study investigates the molecular mechanisms by which IFN-gamma produced as a result of in vivo IL-12 administration exerts its anti-tumor effects. rIL-12 was administered three or five times into mice bearing CSA1M fibrosarcoma, OV-HM ovarian carcinoma or MCH-1-A1 fibrosarcoma. This regimen induced complete regression of CSA1M and OV-HM tumors but only transient growth inhibition of MCH-1-A1 tumors. The anti-tumor effects of IL-12 were associated with enhanced induction of IFN-gamma because these effects were abrogated by pretreatment of hosts with anti-IFN-gamma antibody. Exposure in vitro of the three types of tumor cells to rRFN-gamma resulted in moderate to potent inhibition of tumor cell growth. IFN-gamma stimulated the expression of mRNAs for an inducible type of NO synthase (iNOS) in CSA1M cells and indoleamine 2,3-dioxygenase (IDO), an enzyme capable of degrading tryptophan, in OH-HM cells, but induced only marginal levels of these mRNAs in MCH-1-A1 cells. In association with iNOS gene expression, IFN-gamma-stimulated CSA1M cells produced a large amount of NO which functioned to inhibit their own growth in vitro. Although OV-HM and MCH-1A1 cells did not produce NO, they also exhibited NO susceptibility. Whereas the tumor masses from IL-12-treated CSA1M-bearing or OV-HM-bearing mice induced higher levels of iNOS (for CSA1M) or IDO and iNOS (for OV-HM) mRNAs, the MCH-1-A1 tumor mass expressed lower levels of iNOS mRNA alone. Moreover, massive infiltration of CD4(+) and CD8(+) T cells and Mac-1(+) cells was seen only in the CSA1M and OV-HM tumors. Thus, these results indicate that IFN-gamma produced after IL-12 treatment induces the expression of various genes with potential to modulate tumor cell growth by acting directly on tumor cells or stimulating tumor-infiltrating lymphoid cells and that the effectiveness of IL-12 therapy is associated with the operation of these mechanisms.
Yu
W G
WG
Biomedical Research Center, Osaka University, Medical School, 2-2, Yamada-oka, Suita, Osaka 565, Japan.
Yamamoto
N
N
Takenaka
H
H
Mu
J
J
Tai
X G
XG
Zou
J P
JP
Ogawa
M
M
Tsutsui
T
T
Wijesuriya
R
R
Yoshida
R
R
Herrmann
S
S
Fujiwara
H
H
Hamaoka
T
T
D000818:Animals; D002277:Carcinoma / Q000188:drug therapy / Q000201:enzymology; D002634:Chemotaxis, Leukocyte; D005260:Female; D005354:Fibrosarcoma / Q000188:drug therapy / Q000201:enzymology; D005786:Gene Expression Regulation / Q000276:immunology; D006084:Graft Rejection; D006131:Growth Inhibitors / Q000494:pharmacology*; D016233:Immunotherapy, Active; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase; D007371:Interferon-gamma / Q000494:pharmacology*; D018664:Interleukin-12 / Q000494:pharmacology* / Q000627:therapeutic use*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D009569:Nitric Oxide / Q000502:physiology; D019001:Nitric Oxide Synthase / Q000032:analysis; D010051:Ovarian Neoplasms / Q000188:drug therapy / Q000201:enzymology; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins / Q000494:pharmacology; D014366:Tryptophan Oxygenase / Q000032:analysis; D014407:Tumor Cells, Cultured
D006131:Growth Inhibitors; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase; D012333:RNA, Messenger; D011994:Recombinant Proteins; D018664:Interleukin-12; D009569:Nitric Oxide; D007371:Interferon-gamma; D014366:Tryptophan Oxygenase; D019001:Nitric Oxide Synthase