Detection of HLA class II-dependent T helper antigen using antigen phage display |
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| Authors: | Somasundaram R Satyamoorthy K Caputo L Yssel H Herlyn D |
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| Affiliation: | The Wistar Institute, Philadelphia, PA 19104, USA. |
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| Abstract: | Major histocompatibility complex (MHC) class II-dependent antigens not only activate CD4+ T helper (Th) cells, but also cytolytic T lymphocytes and effector cells of the innate immune system. These antigens therefore are candidate vaccines against cancer and infectious agents. We have developed a novel approach using a model antigen, tetanus toxoid (TT), which provides the basis for the establishment of a novel strategy of cloning Th antigens. In the TT model system, a cDNA library encoding part of the TT light chain which contained a TT-associated Th epitope recognized by TT-specific Th clones was displayed on a phage vector (TT-phage) and presented to TT-specific Th cells by autologous Epstein-Barr virus-transformed B cells (APC). These TT-phages were able to specifically activate two different TT-specific CD4+ Th cell lines as demonstrated both in [3H]thymidine incorporation and cytokine release assays. Th cell stimulation by TT-phages was significant at a ratio of one TT-phage in 50 irrelevant phages. The described approach provides the basis for the development of a novel strategy of cloning MHC class II-dependent Th antigens, using available Th cells. This strategy has several potential advantages over existing antigen cloning methods or biochemical peptide isolation. |
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| Keywords: | phage class II antigens tetanus toxoid antigen cloning antigen presentation |
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