Survivin specific T-cell receptor targeting tumor but not T cells
A cell receptor and survivin technology, applied in the direction of tumor/cancer cells, receptors/cell surface antigens/cell surface determinants, cells modified by introducing foreign genetic material, etc., can solve severe toxicity and other problems
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[0180] The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventors to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of ordinary skill in the art should, in light of the present invention, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or a similar result without departing from the spirit and scope of the invention.
example 1
[0182] Generation of autologous survivin-specific T cell clones with selective antitumor effects
[0183] Use from HLA-A*02 + Peripheral blood samples collected from healthy donors, using survivin 95-104 Survivin 96-104 97M (termed LML; LMLGEFLKL; SEQ ID NO: 16), produces survivin restricted to HLA-A*0201 95-104 (referred to as ELT; ELTLGEFLKL; SEQ ID NO: 15) epitope specific CD8 + Cytotoxic T cells (CTLs) (Andersen et al., 2001).
[0184] After 3 antigen-specific stimulations, 3 of 5 CTL strains (from donors #2, 4, and 5) compared with LML (643±5, 49±1 and 96±7 SFC / 10 5 T cells) and ELT peptides (662±65, 45±6 and 86±9SFC / 10 5 T cells) specific response. By limiting dilution from the most responsive donor (#2) and generating individual T cell clones using multiple assays comparing survivin-specific and non-specific (irrelevant) clones, the optimal functional parent was identified. A joint force. Specifically, clone #24 showed the highest specificity (>99%) for LML-te...
example 2
[0190] Polyclonal T cells engineered to express survivin-specific TCRs are not self-mutilating
[0191] After replacement of the constant regions with the corresponding murine regions, the TCR α and β chains of clone #24 (designated s24-TCR hereafter) were cloned, codon-optimized and encoded into a retroviral vector ( figure 2 A). The TCR chain utilization and complementarity-determining regions are completely different from previously published self-mutilating TCRs (Tables 2 and 3).
[0192] Table 2. Survivin-specific TCR alpha chain utilization.
[0193]
[0194] Nomenclature according to the International Immunogenetics Information System website www.jmgt.org . The sequences of TCRs A66, A71 and A72 are published allotopically restricted survivin-specific TCRs with self-mutilating effects (Leisegang, 2010).
[0195] Table 3. Survivin-specific TCR beta chain utilization.
[0196]
[0197] Nomenclature according to the International Immunogenetics Information Sys...
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