# US 10,500,257 B2 · Method of selecting WT1 vaccine adaptive patient

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,500,257 B2
- Document ID: US10500257B2
- Status: Granted
- Kind: B2
- Application number: 10/562,486
- Filed: 2004-06-25
- Published: 2007-06-07
- Granted: 2019-12-10
- Earliest claimed: 2003-06-27
- Assignees: International Institute of Cancer Immunology, Inc.
- Applicants: Haruo Sugiyama
- Inventors: Haruo Sugiyama
- Web page: https://patentgrep.com/patent/US10500257B2

## CPC

- A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53)
- A61P 35/00: Antineoplastic agents
- G01N 33/505: {involving T-cells}
- G01N 33/5091: {for testing the pathological state of an organism}
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

A method of selecting patients highly responsive to WT1 vaccine and a therapeutic method for treating cancer involving said method of selection are provided, which method of selecting patients highly responsive to WT1 vaccine comprises: (a) isolating a biological sample containing CTL precursor cells from a test subject; (b) measuring the existence frequency or amount of WT1-specific CTL precursor cells in the biological sample of (a); and (c) deciding whether or not the measured value of (b) is high by comparison with that of healthy subject, and evaluating the responsiveness to WT1 vaccine.

## Claims

### Claim 1

A method of selecting a cancer patient highly responsive to WT1 vaccine and treating the patient, comprising:
(a) isolating PBMCs or blood containing CTL precursor cells from a cancer patient who has not been previously immunized with WT1 vaccine;
(b) contacting the PBMCs or blood of (a) with an HLA tetramer comprising a peptide, wherein the HLA tetramer is an HLA-A2 tetramer comprising a peptide comprising the amino acid sequence of SEQ ID NO: 4, or an HLA-A24 tetramer comprising a peptide comprising the amino acid sequence of SEQ ID NO: 2 or 3,
(c) determining the number of CD8+CD45RA+CD27− CTL precursor cells bound to any one of the HLA tetramers relative to the number of CD8+ CTL precursor cells bound to the same HLA tetramer, and
(d) administering a WT1 vaccine to a cancer patient who has a ratio of CD8+CD45RA+CD27− to CD8+ cells as determined in (c) that is significantly higher, as defined by a p value of <0.05, than the ratio of CD8+CD45RA+CD27− CTL precursor cells to CD8+ CTL precursor cells which bind the same tetramer as in (c) contained within the PBMCs or blood of a control subject who does not have cancer,
wherein the WT1 vaccine comprises at least one WT1-derived cancer antigen peptide selected from the group consisting of:
(SEQ ID NO: 2)
Cys Met Thr Trp Asn Gln Met Asn Leu,
(SEQ ID NO: 3)
Cys Tyr Thr Trp Asn Gln Met Asn Leu,
and
(SEQ ID NO: 4)
Arg Met Phe Pro Asn Ala Pro Tyr Leu.

### Claim 2 (depends on 1)

The method of claim 1, wherein the cancer patient has leukemia, myelodysplastic syndrome, multiple myeloma, malignant lymphoma or another hematopoietic malignancy.

### Claim 3 (depends on 1)

The method of claim 1, wherein the cancer patient has lung cancer or another solid cancer.

### Claim 4 (depends on 1)

The method of claim 1, wherein the biological sample is blood.

### Claim 5 (depends on 1)

The method of claim 1, wherein the biological sample comprises PBMCs isolated from blood.

### Claim 6 (depends on 1)

The method of claim 1, wherein the HLA tetramer is the HLA-A2 tetramer comprising the peptide comprising the amino acid sequence of SEQ ID NO: 4.

### Claim 7 (depends on 1)

The method of claim 1, wherein the HLA tetramer is the HLA-A2 tetramer comprising the peptide comprising the amino acid sequence of SEQ ID NO: 2 or 3.

### Claim 8 (depends on 1)

The method of claim 1, wherein (c) comprises:
contacting the PBMCs or blood with the HLA tetramer and with an anti-CD8 antibody, an anti-CD45RA antibody, and an anti-CD27 antibody,
detecting the number of CD8+CD45RA+CD27− CTL precursor cells bound to the HLA tetramer, and
detecting the number of CD8+ CTL precursor cells bound to the HLA tetramer.

### Claim 9 (depends on 1)

The method of claim 1, wherein (c) comprises flow cytometry.

### Claim 10 (depends on 1)

The method of claim 1, wherein in (d) the ratio of CD8+CD45RA+CD27− CTL precursor cells bound to the HLA tetramer in the cancer patient is 1.5 times or higher than the ratio of HLA tetramer-binding CD8+CD45RA+CD27− CTL precursor cells from the control subject who does not have cancer.

### Claim 11 (depends on 1)

The method of claim 1, wherein in (d) the ratio of CD8+CD45RA+CD27− CTL precursor cells bound to the HLA tetramer in the cancer patient is 2.0 times or higher than the ratio of HLA tetramer-binding CD8+CD45RA+CD27− CTL precursor cells from the control subject who does not have cancer.

### Claim 12

A method of selecting a cancer patient highly responsive to a WT1 vaccine and treating the selected patient, comprising:
(a) isolating a PBMCs or blood comprising CTL precursor cells from a cancer patient who has not been previously immunized with a WT1 vaccine;
(b) contacting the PBMCs or blood of (a) with an HLA tetramer comprising a peptide, wherein the HLA tetramer is an HLA-A2 tetramer comprising a peptide comprising the amino acid sequence of SEQ ID NO: 4, or an HLA-A24 tetramer comprising a peptide comprising the amino acid sequence of SEQ ID NO: 2 or 3,
(c) determining the number of CD8+CD45RA+CD27− CTL precursor cells bound to any one of the HLA tetramers relative to the number of CD8+ CTL precursor cells bound to the same HLA tetramer,
wherein the determining in (c) comprises:
contacting the PBMCs or blood with the HLA tetramer and with an anti-CD8 antibody, an anti-CD45RA antibody, and an anti-CD27 antibody, and
(d) identifying a cancer patient who has a ratio of CD8+CD45RA+CD27− to CD8+ cells as determined in (c) that is significantly higher, as defined by a p value of <0.05, than the ratio of CD8+CD45RA+CD27− CTL precursor cells to CD8+ CTL precursor cells which bind the same tetramer as in (c) contained within the PBMCs or blood of a control subject who does not have cancer, and
administering the WT1 vaccine to the identified cancer patient,
wherein the WT1 vaccine comprises at least one WT1-derived cancer antigen peptide selected from the group consisting of:
(SEQ ID NO: 2)
Cys Met Thr Trp Asn Gln Met Asn Leu,
(SEQ ID NO: 3)
Cys Tyr Thr Trp Asn Gln Met Asn Leu,
and
(SEQ ID NO: 4)
Arg Met Phe Pro Asn Ala Pro Tyr Leu.

### Claim 13 (depends on 1)

The method of claim 1, wherein
the cancer is hematopoietic malignancies, lung cancer, or breast cancer;
the HLA tetramer is the HLA-A24 tetramer comprising the peptide comprising the amino acid sequence of SEQ ID NO: 2 or 3; and
the WT1 vaccine comprises at least the WT1-derived cancer antigen peptide of SEQ ID NO: 2 or 3.

### Claim 14 (depends on 12)

The method of claim 12, wherein
the cancer is hematopoietic malignancies, lung cancer, or breast cancer;
the HLA tetramer is the HLA-A24 tetramer comprising the peptide comprising the amino acid sequence of SEQ ID NO: 2 or 3; and
the WT1 vaccine comprises at least the WT1-derived cancer antigen peptide of SEQ ID NO: 2 or 3.

### Claim 15 (depends on 13)

The method of claim 13, wherein
the cancer is acute myelocytic leukemia (AML), acute lymphatic leukemia (ALL), chronic myelocytic leukemia (CML), myelodysplastic syndrome (MDS), lung cancer, or breast cancer.

### Claim 16 (depends on 14)

The method of claim 14, wherein
the cancer is acute myelocytic leukemia (AML), acute lymphatic leukemia (ALL), chronic myelocytic leukemia (CML), myelodysplastic syndrome (MDS), lung cancer, or breast cancer.