Expression and purification of prostate-specific membrane antigen in the baculovirus expression system and recognition by prostate-specific membrane antigen-specific T cells.
Antigen-specific immunotherapy of cancer depends on a consistent source of well-defined protein antigen. Production of recombinant protein offers the obvious solution to this problem but few comparisons of recombinant and native proteins in cellular immune assays have been reported. We report expression of a putative immunotherapy antigen, prostate-specific membrane antigen (PSMA), in insect cells using a baculovirus vector. T cells stimulated with recombinant PSMA or native PSMA derived from the LNCaP cell line recognized both native PSMA and recombinant, baculoviral PSMA. These data indicate that PSMA produced in Sf9 cells is immunologically cross-reactive with native PSMA and therefore suitable for immunotherapy as it is recognized by both cellular and humoral immune responses.
Northwest Biotherapeutics, L.L.C., WA 98125, USA.
P A
PA
Lodge
R A
RA
Childs
S J
SJ
Monahan
J G
JG
McLean
A
A
Sehgal
A L
AL
Boynton
M L
ML
Salgaller
G P
GP
Murphy
D000917:Antibody Formation; D016367:Baculoviridae / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D017252:CD3 Complex / Q000032:analysis / Q000276:immunology; D015704:CD4 Antigens / Q000032:analysis / Q000276:immunology; D016827:CD8 Antigens / Q000032:analysis / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology; D005822:Genetic Vectors; D006801:Humans; D007111:Immunity, Cellular; D007167:Immunotherapy / Q000379:methods; D008297:Male; D017430:Prostate-Specific Antigen / Q000276:immunology* / Q000302:isolation & purification*; D011471:Prostatic Neoplasms / Q000276:immunology* / Q000628:therapy; D014176:Protein Biosynthesis; D011995:Recombination, Genetic; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000276:immunology*; D014407:Tumor Cells, Cultured
D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; D017430:Prostate-Specific Antigen