Inoculation of human interleukin-17 gene-transfected Meth-A fibrosarcoma cells induces T cell-dependent tumor-specific immunity in mice.
OBJECTIVE
The biological activities of interleukin-17 (IL-17), a newly cloned cytokine, have not been fully elucidated. The present study was designed to assess the in vitro and in vivo effect of transfecting the IL-17 gene into tumor cells.


METHODS
A complementary DNA (cDNA) encoding human IL-17 (hIL-17) was obtained by polymerase chain reaction amplification from the human CD4+ T cell cDNA library and inserted into the plasmid pRc/cytomegalovirus to construct an expression vector for the hIL-17 gene. Murine Meth-A fibrosarcoma cells were transfected with the hIL-17 gene using the lipofectin method. The hIL-17 gene-expressing clone (Meth-A/IL-17) was selected and analyzed for cytokine expression by Northern blot.


RESULTS
There was no significant difference in the in vitro proliferation rate among parent Meth-A, cells transfected with vector alone and Meth-A/IL-17 cells. When the tumor cells were transplanted subcutaneously into BALB/c nude (nu+/nu+) mice, there was no difference in in vivo growth rates among the three cell lines. Challenge with tumor cells in conventional BALB/c mice, however, resulted in the rejection of Meth-A/IL-17 cells, but the other two lines did grow. After immunization with Meth-A/IL-17 cells, the mice were rechallenged by parent Meth-A or syngeneic MOPC-104E plasmacytoma cells; the immunized mice rejected the Meth-A cells, but not the MOPC-104E cells. Injecting the anti-thy 1,2 (CD90), anti-CD4 or anti-CD8 monoclonal antibody into conventional BALB/c mice resulted in the resumption of in vivo growth of Meth-A/IL-17 cells, but injecting the anti-asialo GM1 antibody did not. Furthermore, flow cytometric analysis demonstrated a significant increase in the expression of major histocompatibility complex (MHC) class I and class II antigens and lymphocyte function-associated antigen-1 on Meth-A/IL-17 cells.


CONCLUSIONS
Meth-A cells transfected with the hIL-17 gene can induce tumor-specific antitumor immunity by augmenting the expression of MHC class I and II antigens, and both CD4+ and CD8+ T cells may play important roles in inducing antitumor immunity, suggesting the possibility of developing a tumor vaccine incorporating IL-17-transfected tumor cells.
Hirahara
N
N
First Department of Surgery, Shimane Medical University, Izumo, Shimane, Japan.
Nio
Y
Y
Sasaki
S
S
Minari
Y
Y
Takamura
M
M
Iguchi
C
C
Dong
M
M
Yamasawa
K
K
Tamura
K
K
D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D015152:Blotting, Northern; D002455:Cell Division; D004797:Enzyme-Linked Immunosorbent Assay; D005354:Fibrosarcoma / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007111:Immunity, Cellular; D007150:Immunohistochemistry; D007167:Immunotherapy / Q000379:methods*; D020381:Interleukin-17 / Q000235:genetics* / Q000276:immunology*; D007694:Killer Cells, Natural / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
D000951:Antigens, Neoplasm; D020381:Interleukin-17; C032934:Meth A sarcoma-associated rejection antigen, mouse