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-11338032-B2
Publication Date
2022-05-24
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 an isolated population of human papillomavirus (HPV)-specific T cells from HPV-positive tumor samples. The process includes dividing the tumor into multiple fragments, separately culturing these fragments in the presence of at least one cytokine (often IL-2), obtaining T cells from the cultures, testing the T cells for specific autologous HPV-positive tumor recognition and/or HPV antigen recognition, selecting recognizing T cells, and expanding their numbers to produce HPV-specific T cells suitable for adoptive cell therapy.
These methods enable the generation of T cells that specifically recognize HPV antigens E6 and E7, which are constitutively and exclusively expressed by HPV-associated cancer cells, minimizing damage to normal cells and thus reducing toxicity. The methods also allow for nonmyeloablative chemotherapy pretreatment, making them applicable to patients ineligible for total body irradiation due to prior therapies or comorbid conditions.
Furthermore, the inventive methods address the unmet need for new treatments for HPV-associated cancers, which often have poor prognosis despite existing chemotherapy and other therapies. The methods produce HPV-specific T cells at a grade and scale suitable for clinical use, and these cells have been shown capable of targeting HPV-positive cancer cells, including cancers resistant to other treatments such as chemotherapy, surgery, or radiation.
Claims Coverage
The patent presents one independent claim covering a method of treating or preventing HPV-positive cancer by culturing fragmented tumor samples to obtain and expand HPV-specific T cells, followed by administration to the patient.
Culturing multiple fragments of an HPV-positive tumor sample in cytokine presence
The method includes culturing multiple fragments of an HPV-positive tumor sample from the patient in the presence of at least one cytokine, such as IL-2 at 6000 IU.
Obtaining and expanding T cells using feeder cells, OKT3 antibody, and IL-2 without CD4+ depletion
T cells obtained from cultured fragments are expanded in a gas permeable container using irradiated allogeneic and/or autologous feeder cells combined with OKT3 antibody and interleukin-2, notably without depleting CD4+ cells.
Fragmentation of tumor samples by mechanical or enzymatic means
Prior to culturing, the tumor sample is fragmented mechanically or enzymatically to produce multiple tumor fragments for separate culture.
Recognition of specific HPV antigens by expanded T cells
The expanded HPV-specific T cell population recognizes HPV antigens, specifically HPV 16 E6 and E7 or HPV 18 E6 and E7.
Administration of nonmyeloablative lymphodepleting chemotherapy prior to T cell administration
The method may include administering nonmyeloablative lymphodepleting chemotherapy to the patient prior to administration of the expanded HPV-specific T cells.
Expanded T cells comprise multiple T cells with different HPV specificities and secrete interferon-gamma
The expanded T cell population includes multiple T cells with different HPV antigen specificities and secretes at least about 200 pg/mL of interferon-gamma.
Clinical parameters for expansion and administration
Expansion occurs over about 10 to 14 days with a 1000- to 3000-fold increase in T cells. The number of T cells administered ranges between about 1×10^10 to 13.7×10^10, optionally with a pharmaceutically acceptable carrier such as saline or electrolyte solutions.
Enrichment of HPV-specific T cells for CD4+ cells
The expansion can include enrichment for CD4+ T cells prior to administration to the patient.
Overall, the claims cover a method of producing clinically effective HPV-specific T cells by culturing divided tumor fragments with cytokines, expanding T cells with feeder cells combined with OKT3 antibody and IL-2 without CD4+ cell depletion, selecting for HPV antigen recognition, and administering the expanded T cells following lymphodepleting chemotherapy to treat or prevent HPV-positive cancers.
Stated Advantages
Ability to generate HPV-specific T cells at a clinical grade and scale appropriate for therapy.
Selective targeting of HPV antigens E6 and E7, expressed only on cancer cells, thus minimizing damage to normal cells and reducing toxicity.
Applicability to patients ineligible for total body irradiation due to prior myeloablative therapies or comorbid conditions via use of nonmyeloablative chemotherapy.
Effective treatment for HPV-positive cancers that are resistant to other therapies such as chemotherapy, surgery, or radiation.
Documented Applications
Adoptive cell therapy for treating or preventing HPV-positive cancers, including cervical cancer and head and neck squamous cell carcinoma.
Generation of HPV-specific T cells for use in therapeutic regimens involving administration following nonmyeloablative lymphodepleting chemotherapy.
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