A two-component modular approach for enhancing T-cell activation utilizing a unique anti-FcgammaRI-streptavidin construct and microspheres coated with biotinylated-antigen.
The professional antigen presenting cell (APC) plays an essential role in the initiation and propagation of the acquired immune response. Thus, much work has been done in designing strategies that target vaccine antigen (Ag) to APC. Utilizing recombinant DNA technology, we have created a unique two-component system that delivers biotinylated Ag to the Fc gamma receptor type I (FcgammaRI) on APC. Our studies demonstrate that we can successfully engineer FcgammaRI-specific targeting element proteins that simultaneously bind both biotin and recognize FcgammaRI. Additionally, we are able to engineer biotinylated Ag, which form functional elements when adsorbed onto latex microspheres. Furthermore, the targeting and functional element components bind to each other and successfully form two-component immunogens. T-cell activation in response to targeted Ag-laden microspheres is 10- to 100-fold greater than the response to the non-targeted Ag-laden microspheres. This enhancement is 100- to 1000-fold greater than the responses generated to soluble Ag. Thus, our results suggest that specific targeting of Ag-laden microspheres to FcgammaRI may significantly enhance the adjuvant properties of microparticulate delivery systems. Further development of this system may help to elucidate the mechanisms involved in generating enhanced responses to APC-targeted vaccines and significantly advance vaccine technology.
Walsh
Mary C
MC
Center for Immunology and Microbial Disease, MC-151, Albany Medical College, 47 New Scotland Avenue, Albany, NY 12201, USA.
Banas
Jeffrey A
JA
Mudzinski
Stanley P
SP
Preissler
Mark T
MT
Graziano
Robert F
RF
Gosselin
Edmund J
EJ
D000938:Antigen-Presenting Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D000941:Antigens; D019921:Biotinylation / Q000379:methods; D002450:Cell Communication / Q000502:physiology; D002478:Cells, Cultured; D004338:Drug Combinations; D016503:Drug Delivery Systems / Q000379:methods*; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects*; D008863:Microspheres; D011786:Quality Control; D017452:Receptors, IgG / Q000008:administration & dosage* / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D019809:Streptavidin / Q000008:administration & dosage* / Q000276:immunology / Q000378:metabolism; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D020298:U937 Cells
D000941:Antigens; D004338:Drug Combinations; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins; D019809:Streptavidin