Induction of antitumor response by in vivo allogeneic major histocompatibility complex gene transfer using electroporation.
BACKGROUND
Most malignancies lack tumor-associated antigens, which are recognized by T-lymphocytes through major histocompatibility complex (MHC) molecules, and seem to evade host immunological defense. In this study, we investigated whether allogeneic MHC gene transfer into the low-antigenic tumor might induce cell-mediated immunity, and provide an immunotherapeutic effect in vivo.


METHODS
Mammary adenocarcinoma cells (MAT B III) originated from an F344 rat (RT1A(l)) were transfected with a plasmid DNA encoding RT1A(a) (pcMRT1A) in vitro. Cytolytic T-cell response was then evaluated using F344 splenocytes. Furthermore, pcMRT1A-liposome complex was injected into the MAT B III tumors grown in F344 rats, followed by a series of eight electrical pulses with a electroporator, every other d for total of four times. The tumor size and survival of animals were evaluated thereafter.


RESULTS
The expression of RT1A(a) on the tumor cells induced cytolytic T-cell response in vitro. Intratumor injection of pcMRT1A-liposome complex followed by in vivo electroporation markedly generated biological response, including increased interferon-gamma (IFN-gamma) expression and a large number of infiltrated cells in the tumor. Inhibited tumor growth, even complete tumor regression, was observed, resulting in prolonged survival time.


CONCLUSIONS
Direct transfer of allogeneic MHC gene into the tumor by in vivo electroporation could induce a cell-mediated immune response against the introduced antigens, resulting in local IFN-gamma production. This initial response may bring about the subsequent immunological response even to the unmodified tumor cells through cytokines such as IFN-gamma, resulting in the marked tumor regression.
Shimizu
Hiroaki
H
Department of General Surgery, Graduate School of Medicine, Chiba University, Chiba, Japan. h-shimizu@umin.ac.jp
Nukui
Yuji
Y
Mitsuhashi
Noboru
N
Kimura
Fumio
F
Yoshidome
Hiroyuki
H
Ohtsuka
Masayuki
M
Kato
Atsushi
A
Miyazaki
Masaru
M
D000818:Animals; D018274:Electroporation / Q000379:methods*; D005260:Female; D018014:Gene Transfer Techniques; D008285:Major Histocompatibility Complex / Q000235:genetics*; D015674:Mammary Neoplasms, Animal / Q000235:genetics* / Q000401:mortality / Q000628:therapy*; D009419:Nerve Tissue Proteins / Q000235:genetics; D051381:Rats; D011916:Rats, Inbred F344; D016019:Survival Analysis; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection
D009419:Nerve Tissue Proteins; C477690:Snx27 protein, rat