Peptide fine specificity of anti-glycoprotein 100 CTL is preserved following transfer of engineered TCR alpha beta genes into primary human T lymphocytes
TCR with known antitumor reactivity can be genetically introduced into primary human T lymphocytes and provide promising tools for immunogene therapy of tumors. We molecularly characterized two distinct TCRs specific for the same HLA-A2-restricted peptide derived from the melanocyte differentiation Ag gp100, yet exhibiting different stringencies in peptide requirements. The existence of these two distinct gp100-specific TCRs allowed us to study the preservation of peptide fine specificity of native TCRalphabeta when engineered for TCR gene transfer into human T lymphocytes. Retroviral transduction of primary human T lymphocytes with either one of the two sets of TCRalphabeta constructs enabled T lymphocytes to specifically kill and produce TNF-alpha when triggered by native gp100(pos)/HLA-A2(pos) tumor target cells as well as gp100 peptide-loaded HLA-A2(pos) tumor cells. Peptide titration studies revealed that the cytolytic efficiencies of the T lymphocyte transductants were in the same range as those of the parental CTL clones. Moreover, primary human T lymphocytes expressing either one of the two engineered gp100-specific TCRs show cytolytic activities in response to a large panel of peptide mutants that are identical with those of the parental CTL. The finding that two gp100-specific TCR, derived from two different CTL, can be functionally introduced into primary human T lymphocytes without loss of the Ag reactivity and peptide fine specificity, holds great promise for the application of TCR gene transfer in cancer immunotherapy.
|Keywords||Amino Acid Sequence, Amino Acid Substitution/genetics/immunology, Base Sequence, Cell Line, Cell Line, Transformed, Cells, Cultured, Clone Cells, Cytotoxicity Tests, Immunologic, Epitopes, T-Lymphocyte/genetics/*immunology, HLA-A2 Antigen/immunology, Humans, K562 Cells, Melanoma/immunology/therapy, Membrane Glycoproteins/genetics/*immunology, Molecular Sequence Data, Neoplasm Proteins/genetics/*immunology, Peptide Fragments/genetics/*immunology, Receptors, Antigen, T-Cell, alpha-beta/biosynthesis/*genetics/therapeutic use, Research Support, Non-U.S. Gov't, T-Lymphocyte Subsets/*immunology/metabolism, T-Lymphocytes, Cytotoxic/*immunology/metabolism, Transduction, Genetic/*methods, Transfection, Tumor Cells, Cultured|
Schaft, N., Willemsen, R.A., de Vries, J., Lankiewicz, B., Essers, B.W., Gratama, J.W., … Bolhuis, R.L.H.. (2003). Peptide fine specificity of anti-glycoprotein 100 CTL is preserved following transfer of engineered TCR alpha beta genes into primary human T lymphocytes. Journal of Immunology. Retrieved from http://hdl.handle.net/1765/10098