Means and methods for active cellular immunotherapy of cancer by using tumor cells killed by high hydrostatic pressure and dendritic cells
Inventors
BARTUNKOVÁ JIRINA, null • Bartů{hacek over (n)}ková, Ji{hacek over (r)}ina • ŠPÍŠEK RADEK, null • {hacek over (S)}PÍ{hacek over (S)}EK, Radek
Assignees
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Publication Number
US-10918704-B2
Publication Date
2021-02-16
Expiration Date
Abstract
Disclosed are pharmaceutical compositions for inducing an immune response against tumor cells comprising tumor cells which are made apoptotic by treatment with high hydrostatic pressure and dendritic cells, and methods for producing such compositions.
Core Innovation
The invention relates to a method for producing a mature loaded dendritic cell for cancer active cellular immunotherapy. The method obtains monocytes or immature dendritic cells from a patient, and culturing or providing immature dendritic cells that are further loaded with apoptotic tumor cells. The tumor cells are derived from the patient or from one or more tumor cell lines, and apoptosis is induced by applying high hydrostatic pressure (HHP) to the tumor cells.
Apoptosis induced by applying HHP of 100 MPa to 300 MPa for 10 min to 2 h produces tumor cells that are loaded in vitro onto immature dendritic cells. The loading is carried out by combining immature dendritic cells with apoptotic tumor cells at a ratio between about 1:1 to about 10:1, to obtain loaded dendritic cells. The loaded dendritic cells are then further matured in vitro by treating with Poly I:C or LPC to obtain loaded matured dendritic cells.
The disclosed approach is used to generate immunogenic cell death activity in the tumor cells without additional adjuvant, including release of HMGB1 and expression of immunogenic cell death markers. The loaded matured dendritic cells are characterized by maturation and activation markers such as CD86 and CD83 or HLA-DR, and by induction of phagocytosis. The document further reports that tumor-specific CD4/CD8 T-cell responses are induced while regulatory T-cell expansion is reduced, and that the anti-tumor response is stronger than immunogenic tumor cells alone.
Claims Coverage
The document contains two independent methods with overlapping core steps, each including HHP-induced apoptosis of tumor cells, in vitro loading onto immature dendritic cells at a defined ratio, and further in vitro maturation using Poly I:C or an alternative maturation agent. Across the independent claims, the main inventive features include HHP-driven generation of apoptotic tumor cells, in vitro loading of immature dendritic cells with those apoptotic tumor cells, and producing mature loaded dendritic cells by Poly I:C (or LPC/LPS) treatment.
HHP-induced apoptotic tumor cells loaded onto immature dendritic cells
Inducing apoptosis in tumor cells by applying high hydrostatic pressure (HHP) of 100 MPa to 300 MPa for 10 min to 2 h, and loading in vitro the immature dendritic cells with the apoptotic tumor cells to obtain loaded dendritic cells.
Defined immature dendritic cell to tumor cell ratio for loading
Combining the immature dendritic cells with the apoptotic tumor cells at a ratio between about 1:1 to about 10:1.
In vitro maturation of loaded dendritic cells with Poly I:C (and alternative agents)
Further maturing in vitro the loaded dendritic cells by treating the loaded dendritic cells with Poly I:C or LPC (or Poly I:C or LPS in the other independent claim) to obtain loaded matured dendritic cells.
GM-CSF and IL-4 culture to obtain immature dendritic cells from monocytes
Culturing monocytes in the presence of GM-CSF and IL-4 to obtain immature dendritic cells.
Overall, the claim coverage centers on producing mature loaded dendritic cells by generating HHP-apoptotic tumor cells, loading them onto immature dendritic cells at a specified ratio, and completing maturation in vitro using Poly I:C and an additional specified maturation agent (LPC or LPS). One independent claim further requires obtaining immature dendritic cells by culturing patient monocytes with GM-CSF and IL-4.
Stated Advantages
Induces robust tumor-specific CD4/CD8 T-cell responses.
Reduced regulatory T-cell expansion.
Stronger anti-tumor response (reported as about ~10-fold stronger) versus immunogenic tumor cells alone.
HHP-killed tumor cells become immunogenic without additional adjuvant.
Matured loaded dendritic cells show increased activation/maturation markers such as CD86 and/or HLA-DR.
Documented Applications
Cancer active cellular immunotherapy using mature loaded dendritic cells.
Prostate cancer, including reported prolongation of PSA doubling time and improved median survival in late-stage/castration-resistant metastatic prostate cancer, including with or alongside docetaxel.
Ovarian cancer.
Acute lymphoblastic leukemia (ALL).
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