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-11376318-B2
Publication Date
2022-07-05
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 relates to methods of preparing an isolated population 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 cultured fragments, testing for specific autologous HPV-positive tumor recognition or HPV antigen recognition, selecting such T cells, and expanding their number to produce a population of HPV-specific T cells for adoptive cell therapy. The methods also relate to treating or preventing cancer using these T cells.
The problem being addressed is the poor prognosis of many cancers, including HPV-associated cancers, despite advances in treatments such as chemotherapy. There exists an unmet need for additional treatments for cancer, particularly HPV-associated cancers.
The inventive methods enable generation of T cells at a grade and scale suitable for clinical use that specifically recognize HPV antigens E6 and E7, which are constitutively and specifically expressed by cancer cells but not by normal cells. This targeting potentially reduces toxicity by minimizing destruction of normal cells. The methods include culturing tumor fragments in the presence of only one cytokine (preferably IL-2), and expanding the selected T cells using OKT3 antibody and interleukin-2 with irradiated feeder cells. The invention also includes using nonmyeloablative lymphodepleting chemotherapy prior to T cell administration to treat patients not eligible for total body irradiation. The methods have shown efficacy in treating or preventing HPV-positive cancers that are refractory to existing treatments.
Claims Coverage
The patent includes a number of independent claims primarily related to a tumor infiltrating lymphocyte (TIL) composition and methods of treating HPV-positive cancers using said TILs.
HPV-specific TIL composition production method
A method comprising culturing multiple fragments of an HPV-positive tumor sample in the presence of at least one cytokine, obtaining T cells, expanding the number of T cells using irradiated allogeneic and/or autologous feeder cells combined with OKT3 antibody and interleukin-2 (IL-2), without depleting CD4+ cells, optionally followed by a second culturing step, to produce a therapeutic population of HPV-specific TILs comprising a pharmaceutically acceptable carrier.
Use of IL-2 at specific steps and concentrations
Culturing multiple tumor fragments in the presence of IL-2, preferably at about 6000 IU in the initial culturing step and in the expansion step.
Fragmentation of HPV-positive tumor sample
Obtaining the tumor sample and fragmenting it mechanically or enzymatically prior to culturing.
Therapeutic recognition specificity of expanded TILs
Expanded population of HPV-specific T cells recognizes HPV type 16-positive cervical cancer cells or head and neck squamous cell carcinoma and specifically recognizes HPV antigens including HPV 16 E6 and HPV 16 E7.
Administration method for treating HPV-positive cancer
Administering the TIL composition to a patient via intraarterial, intravenous, intraperitoneal, intrathecal, or intralymphatic routes, optionally following nonmyeloablative lymphodepleting chemotherapy.
Expansion protocol with specific fold increase and duration
Expanding HPV-specific T cells over about 10 to 14 days with a population increase of about 1000- to 3000-fold prior to administration.
Dose range of administered HPV-specific T cells
Administering from about 1×10^10 to about 13.7×10^10 HPV-specific T cells to the patient.
Composition features relating to T cell subpopulations and carrier
Composition can be enriched for CD4+ T cells; the pharmaceutically acceptable carrier includes saline, 5% dextrose in water, Ringer's lactate, electrolyte solutions, or PLASMA-LYTE A; and the initial culturing step is at least about 12 days.
The independent claims cover compositions of HPV-specific TIL expanded using defined culture and expansion protocols and pharmaceutical carriers, as well as methods for treating HPV-positive cancers using these TILs, emphasizing cytokine use, fragmentation methods, TIL reactivity, administration routes, expansion parameters, dosing, and cell population specifics.
Stated Advantages
The inventive methods generate T cells at a clinical grade and scale suitable for use.
The generated T cells specifically recognize HPV antigens E6 and E7, which are expressed by cancer cells but not by normal cells, minimizing toxicity.
The method can be used with nonmyeloablative chemotherapy to treat patients ineligible for total body irradiation.
The methods may successfully treat or prevent HPV-positive cancers resistant to existing therapies such as chemotherapy, surgery, or radiation.
Documented Applications
Treatment or prevention of HPV-positive cancers, including cervical cancer and head and neck squamous cell carcinomas.
Use of the expanded HPV-specific T cells for adoptive cell therapy in mammals, especially humans.
Treatment of cancers associated with HPV 16 or HPV 18 infections.
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