MHC class II-independent, Vbeta-specific activation of T cells by superantigen mutants fused to anti-tumor Fab fragments: implications for use in treatment of human colon carcinoma.
Genetically engineered fusion proteins of the super-antigen staphylococcal enterotoxin A (SEA) and tumor-reactive monoclonal antibodies, C215Fab-SEA and C242Fab-SEA, have been generated and shown to be effective in mediating superantigen-antibody directed cellular cytotoxicity against human carcinoma cells expressing the CA215 or CA242 antigens in an MHC class II-independent manner. In an attempt to reduce the in vivo toxicity of superantigen administration, alanine substitution mutations in SEA at residues F47 and D227 that affect SEA binding to class II molecules have been created and genetically linked to C215Fab or C242Fab. The purpose of this study was to determine whether these Fab-SEA mutant fusion proteins, that have low MHC class II binding affinities, were still able to stimulate human T cells in a Vbeta-specific manner in the presence or absence of MHC class II molecules. The SEA wt- and SEA-D227A-based fusion proteins shared the ability to activate V beta5. 2-, Vbeta6-, Vbeta7-, Vbeta9- and Vbeta18-bearing T cells, whereas Fab-SEA-F47A protein activated only Vbeta6- and Vbeta7-bearing T cells. The fusion of Fab fragments onto SEA wt, SEA-F47A or SEA-D227A had no effect on the Vbeta specificity of these superantigens. Fab fusion proteins containing either SEA wt or SEA mutants were presented, in the absence of class II molecules, by CHO cells transfected with CA215 and CD80 and all induced the expansion of only Vbeta6-, Vbeta7- and Vbeta 18-bearing T cells. Fab-SEA mutant fusion proteins may provide attenuated therapeutic agents that, while still able to specifically target high affinity T cells for MHC class II-independent local tumor killing, will not induce excessive systemic toxicity.
Newton
D W
DW
Departments of Surgery, Microbiology and Immunology, University of Tennessee-Memphis, Memphis, TN 38163, USA.
Dohlsten
M
M
Lando
P A
PA
Kalland
T
T
Olsson
C
C
Kotb
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D016466:CHO Cells; D003110:Colonic Neoplasms / Q000628:therapy*; D006224:Cricetinae; D004768:Enterotoxins / Q000235:genetics / Q000276:immunology*; D015804:HLA-DR4 Antigen / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology*; D008213:Lymphocyte Activation; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013211:Staphylococcus aureus / Q000276:immunology*; D018089:Superantigens / Q000235:genetics / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*
D000911:Antibodies, Monoclonal; D004768:Enterotoxins; D015804:HLA-DR4 Antigen; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011993:Recombinant Fusion Proteins; D018089:Superantigens; C026066:enterotoxin A, Staphylococcal