Gene transfer of the costimulatory molecules B7-1 and B7-2 into human multiple myeloma cells by recombinant adeno-associated virus enhances the cytolytic T cell response.
Gene transfer of the costimulatory molecules B7-1 and B7-2 induces a potent antitumor immune response in a variety of tumor models. B cell neoplasms including multiple myeloma (MM) often show little or no expression of B7 antigens; they are therefore a potential target for this approach. To increase the expression of human B7 genes in MM cells, both genes and the neomycin phosphotransferase gene were packaged into recombinant adeno-associated virus vectors (rAAV). The resulting recombinant viruses rAAV/B7-1, rAAV/B7-2 and rAAV/Neo were used to transduce the MM cell lines LP-1 and RPMI 8226. This allowed the transduction of up to 80% of LP-1 cells 4 days after infection with purified rAAV particles. The response of human allogeneic T cells to rAAV/B7-1 and rAAV/B7-2 transduced, gamma-irradiated LP-1 cells was assessed by [3H]thymidine incorporation, by RT-PCR-based detection of immunostimulatory cytokine transcripts and by ELISA quantification of cytokines in the supernatant. Stimulation of T cells with rAAV/B7-1 or rAAV/B7-2 transduced LP-1 cells resulted in an up to 10-fold increase of T cell proliferation when compared with LP-1 cells transduced with rAAV/Neo. Similar results were obtained with RPMI 8226 cells. Both rAAV/B7-1 and rAAV/B7-2 transduced LP-1 cells stimulated the T cell secretion of IL-2 and IFN-gamma. Furthermore, [51Cr] release assays showed that rAAV/B7-1 or rAAV/B7-2 transduced LP-1 cells induced a cytolytic T cell (CTL) response, in contrast to LP-1 cells transduced with rAAV/Neo. In all assays, the effects of rAAV/B7-1 and rAAV/B7-2 were similar. Taken together, the results show that rAAV-mediated transfer of B7 genes into MM cell lines is able to enhance the antitumor T cell response and to elicit a cytolytic T cell response.
Wendtner
C M
CM
Medizinische Klinik, Klinikum Innenstadt, Germany.
Nolte
A
A
Mangold
E
E
Buhmann
R
R
Maass
G
G
Chiorini
J A
JA
Winnacker
E L
EL
Emmerich
B
B
Kotin
R M
RM
Hallek
M
M
D015703:Antigens, CD / Q000235:genetics*; D018122:B7-1 Antigen / Q000235:genetics*; D051940:B7-2 Antigen; D002478:Cells, Cultured; D003131:Combined Modality Therapy; D000229:Dependovirus; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007167:Immunotherapy; D007376:Interleukin-2 / Q000378:metabolism; D019868:Kanamycin Kinase; D008213:Lymphocyte Activation; D008562:Membrane Glycoproteins / Q000235:genetics*; D009101:Multiple Myeloma / Q000628:therapy*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
D015703:Antigens, CD; D018122:B7-1 Antigen; D051940:B7-2 Antigen; C497964:CD86 protein, human; D007376:Interleukin-2; D008562:Membrane Glycoproteins; D017853:Phosphotransferases (Alcohol Group Acceptor); D019868:Kanamycin Kinase