Receptor gene for peptide cancer antigen-specific T cell
The invention provides the nucleotide sequence and amino acid sequence of the CDR3 domain of the T cell receptor (TCR) gene of a WT1-specific cytotoxic T cell (CTL) against WT1 protein. Also provided are a method for testing for and treating cancer using the nucleotide sequence and amino acid sequence, and a chip, primer set, kit, and device for testing for cancer comprising the nucleotide sequence and amino acid sequence.
1. A method of assaying a clonality of WT1-specific cytotoxic T-cells (CTLs), the method comprising:
(a) obtaining a sample from an HLA-A*0201-positive subject, wherein the sample is a tissue affected with a cancer which is generated from cells containing WT1, a body fluid, or a mucosal membrane;
(b) sorting a single WT1-specific CTL in the sample;
(c) amplifying a polynucleotide from the single WT1-specific CTL, the polynucleotide comprising:
(i) a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-756,
(ii) a complementary RNA sequence of the nucleotide sequence (i), or
(iii) a complementary DNA sequence of the nucleotide sequence (i);
or
hybridizing a polynucleotide in the single WT1-specific CTL with at least one polynucleotide comprising:
(i) a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-756,
(ii) a complementary RNA sequence of the nucleotide sequence (i), or
(iii) a complementary DNA sequence of the nucleotide sequence (i); and
(d) measuring the clonality of WT1-specific CTLs that comprise an identical nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-756, thereby characterizing the WT1-specific CTLs as having a multiplicity of clonality if the clonality is greater than 1,
and/or
measuring the number of types of WT1-specific CTLs having the multiplicity of clonality.
2. The method of claim 1 ,
wherein the at least one polynucleotide in the hybridizing is immobilized on a chip.
3. The method of claim 1 , wherein (c) comprises amplifying a polynucleotide from the single WT1-specific CTL.
4. The method of claim 1 , wherein (c) comprises hybridizing a polynucleotide in the single WT1-specific CTL.
5. The method of claim 1 , wherein (d) comprises measuring the clonality of WT1-specific CTLs that comprise an identical nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-756.
6. The method of claim 1 , wherein (d) comprises measuring the number of types of WT1-specific CTLs having the multiplicity of clonality.
7. A method for assaying a clonality of WT1-specific CTLs, comprising:
(a) obtaining a sample from an HLA-A*0201-positive subject, wherein the sample is a tissue affected with a cancer which is generated from cells containing WT1, a body fluid, or a mucosal membrane;
(b) sorting a single WT1-specific CTL in the sample;
(c) contacting a peptide from a single WT1-specific CTL with an antibody against a peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 757-1512; and
(d) measuring the clonality of WT1-specific CTLs that comprise an identical amino acid sequence selected from the group consisting of SEQ ID NOs: 757-1512, thereby characterizing the WT1-specific CTLs as having a multiplicity of clonality if the clonality is greater than 1;
and/or
measuring the number of types of WT1-specific CTLs that have a multiplicity of clonality.
8. The method of claim 7 , wherein the antibody in the contacting is immobilized on a chip.