Exogenous antigen targeted to FcgammaRI on myeloid cells is presented in association with MHC class I.
Vaccine therapy is attractive for prostate cancer patients because the tumor is slow growing (allowing time to augment host responses) and occurs in an older population less likely to tolerate more toxic treatments. We have constructed an expression vector based on a monoclonal antibody (mAb) that targets the high affinity receptor for IgG (FcgammaRI, CD64) which is exclusively expressed on myeloid cells including dendritic cells (DC). The heavy chain of mAb H22 CH2 and CH3 domains were removed and replaced with the gene for prostate specific antigen (PSA). Using that vector, we have constructed and purified FPH22.PSA, a fusion protein that targets PSA to FcgammaRI on antigen presenting cells (APC). This fusion protein has an apparent molecular mass of 80-83 kDa, binds to FcgammaRI with high affinity and expresses PSA. We demonstrate that FPH22.PSA targeted PSA was internalized and processed by the human myeloid THP-1 cell line resulting in presentation of MHC class I-associated PSA peptides and lysis of THP-1 by PSA-specific human CTL. Moreover, pretreatment of THP-1 cells with antibodies to block either FcgammaRI or MHC class I, blocked lysis indicating that targeting to FcgammaRI results in presentation of exogenous antigen on MHC class I molecules. These data demonstrate that FPH22.PSA was processed in such a manner by the myeloid cell line to allow for presentation of immunodominant peptides in MHC class I molecules and suggests that uptake of antigen via FcgammaRI results in cross-priming.
Department of Microbiology, HB7556, Dartmouth Medical School, 1 Medical Center Drive, Lebanon, NH 03756, USA. pkw@dartmouth.edu
P K
PK
Wallace
K Y
KY
Tsang
J
J
Goldstein
P
P
Correale
T M
TM
Jarry
J
J
Schlom
P M
PM
Guyre
M S
MS
Ernstoff
M W
MW
Fanger
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D017951:Antigen Presentation; D019496:Cancer Vaccines / Q000276:immunology; D003713:Dendritic Cells / Q000502:physiology*; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D017430:Prostate-Specific Antigen / Q000276:immunology* / Q000378:metabolism; D017452:Receptors, IgG / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D019496:Cancer Vaccines; D015395:Histocompatibility Antigens Class I; D007140:Immunoglobulin Fab Fragments; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins; D017430:Prostate-Specific Antigen