Methods of preparing anti-human papillomavirus antigen T cells
Inventors
Hinrichs, Christian S. • Rosenberg, Steven A.
Assignees
US Department of Health and Human Services
Publication Number
US-11077182-B2
Publication Date
2021-08-03
Expiration Date
2034-07-14
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Abstract
Disclosed are methods of preparing an isolated population of human papillomavirus (HPV)-specific T cells comprise dividing an HPV-positive tumor sample into multiple fragments; separately culturing the multiple fragments; obtaining T cells from the cultured multiple fragments; testing the T cells for specific autologous HPV-positive tumor recognition; selecting the T cells that exhibit specific autologous HPV-positive tumor recognition; and expanding the number of selected T cells to produce a population of HPV-specific T cells for adoptive cell therapy. Related methods of treating or preventing cancer using the T cells are also disclosed.
Core Innovation
The invention provides methods of preparing isolated populations of human papillomavirus (HPV)-specific T cells by dividing an HPV-positive tumor sample into multiple fragments, separately culturing these fragments, obtaining T cells from the cultures, testing the T cells for specific autologous HPV-positive tumor recognition and/or HPV antigen recognition, selecting T cells that exhibit such specific recognition, and expanding the selected T cells to produce a population of HPV-specific T cells suitable for adoptive cell therapy.
The problem being addressed is the unmet need for additional treatments for cancers caused by HPV infection, such as uterine cervical cancer, which often have poor prognoses despite advances in chemotherapy and other treatments.
The inventive methods advantageously generate T cells at a grade and scale suitable for clinical use that recognize HPV antigens E6 and E7, which are constitutively expressed by cancer cells but not normal cells, thereby targeting cancer cell destruction while minimizing toxicity to normal cells. The methods include use of nonmyeloablative chemotherapy, enhancing applicability to patients ineligible for total body irradiation-based treatments, and have potential to successfully treat or prevent HPV-positive cancers that are resistant to chemotherapy, surgery, or radiation.
Claims Coverage
The patent includes one independent claim defining a method for treating or preventing HPV-positive cancer using cultured HPV-specific T cells obtained and expanded from fragmented tumor samples.
Culturing HPV-positive tumor fragments in presence of cytokines
The method comprises culturing multiple fragments of an HPV-positive tumor sample from the patient with at least one cytokine.
Obtaining and expanding HPV-specific T cells
T cells are obtained from the cultured fragments and expanded to produce an HPV-specific T-cell population using irradiated allogenic and/or autologous feeder cells, OKT3 antibody, and interleukin-2, without depleting CD4+ cells.
Optional second culturing step
An optional additional culturing step can be added to further expand selected T cells after the primary expansion.
Use of IL-2 cytokine and parameters for culturing and expansion
IL-2 is the preferred cytokine at 6000 IU concentration during culturing and expansion, with expansion occurring over about 10 to 14 days achieving 1000- to 3000-fold cell increase.
Tumor fragmentation methods
Prior to culturing, the HPV-positive tumor sample is obtained and fragmented mechanically or enzymatically.
Targeting specific HPV antigens and cancer types
The expanded T cells recognize HPV antigens HPV 16 E6, HPV 16 E7, HPV 18 E6, and HPV 18 E7, and recognize HPV 16-positive cervical cancer cells; applicable to cancers such as cervical cancer and head and neck squamous cell carcinoma.
Administration and dosing
Administering the expanded HPV-specific T cells to the patient in an amount effective to treat or prevent HPV-positive cancer, with doses ranging approximately from 1×10^10 to 13.7×10^10 cells, optionally with a pharmaceutically acceptable carrier such as saline or electrolyte solutions.
The claims cover a comprehensive method of preparing and expanding HPV-specific T cells from fragmented HPV-positive tumor samples using cytokine culture and specific expansion protocols, followed by administration of the expanded HPV-specific T cells to treat or prevent HPV-positive cancers, particularly cervical and head and neck cancers.
Stated Advantages
The methods generate HPV-specific T cells at clinical grade and scale suitable for adoptive cell therapy.
They produce T cells that specifically recognize constitutively expressed HPV antigens E6 and E7, which target cancer cells while minimizing or eliminating toxicity to normal cells.
Inclusion of nonmyeloablative chemotherapy expands the patient population eligible for treatment by avoiding total body irradiation, allowing treatment of patients with prior myeloablative treatments or comorbidities.
The methods may successfully treat or prevent HPV-positive cancers refractory to chemotherapy, surgery, or radiation.
Documented Applications
Preparation of HPV-specific T cells for adoptive cell therapy to treat or prevent HPV-positive cancers.
Treatment of HPV-associated cancers including cervical cancer, head and neck squamous cell carcinoma, and other HPV-positive cancers.
Use in patients undergoing nonmyeloablative lymphodepleting chemotherapy prior to T cell administration.
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