Therapeutic pooled blood apoptotic cell preparations and uses thereof
Cell preparations comprising a pooled and enriched, mononuclear apoptotic cell population, and a method of preparing this cell preparation are described. The pooled mononuclear apoptotic cell preparation may be obtained from pooled, allogeneic white blood cell fractions that are pooled prior to or following induction of apoptosis. Further, described herein are methods of use of these pooled apoptotic cell preparations for treating an immune disease, an inflammatory disease, an autoimmune disease, or infertility in a subject. For example, a pooled apoptotic cell preparation may be used to treat graft versus host disease (GVHD) in an allogeneic subject.
1. A therapeutic irradiated and pooled, mononuclear, early apoptotic cell preparation comprising individual mononuclear-enriched cell populations in an early apoptotic state that are irradiated post induction of apoptosis;
and wherein said irradiated and pooled, mononuclear-enriched, early apoptotic cell preparation comprises
(a) a decreased number of non-quiescent non-apoptotic cells;
(b) a suppressed cellular activation of any living non-apoptotic cells; or
(c) a reduced proliferation of any living non-apoptotic cells;
or any combination thereof compared with a preparation comprising non-irradiated cell populations.
2. The therapeutic irradiated and pooled early apoptotic cell preparation of claim 1 , wherein said individual mononuclear-enriched populations are pooled prior to induction of apoptosis or post induction of apoptosis and are obtained from a single donor or from multiple donors.
3. The therapeutic irradiated and pooled early apoptotic cell preparation of claim 1 , wherein said pooled, individual, mononuclear cell populations comprise
(a) populations pooled independent of HLA matching of said individual mononuclear cell populations' HLA markers; or ; or
(b) at least one cell type selected from the group consisting of: lymphocytes, monocytes, dendritic cells, and natural killer cells; or
(c) cells comprising inactive T cell receptors or reduced immune activity; or
any combination thereof.
4. The therapeutic irradiated and pooled early apoptotic cell preparation of claim 1 , wherein said mononuclear cell populations comprise cells obtained from between about 2 and 25 units of blood.
5. The therapeutic irradiated and pooled early apoptotic cell preparation of claim 4 , wherein said blood comprises white blood cell (WBC) fractions from blood donations.
6. The therapeutic irradiated and pooled early apoptotic cell preparation of claim 1 , wherein said mononuclear cell populations comprise allogeneic cells from HLA matched or HLA unmatched sources, with respect to a recipient subject.
7. The therapeutic irradiated and pooled early apoptotic cell preparation of claim 1 , wherein said irradiation comprises gamma irradiation or UV irradiation.
8. The therapeutic irradiated and pooled early apoptotic cell preparation of claim 1 , wherein said irradiated, early apoptotic cell populations comprise a decreased percent of non-quiescent non-apoptotic cells per population compared with a non-irradiated cell population.
9. A pharmaceutical composition, comprising the therapeutic irradiated and pooled early apoptotic cell preparation of claim 1 .
10. A method of treating, preventing, ameliorating, inhibiting, or reducing the incidence of an immune disease, an autoimmune disease, a cytokine release syndrome (CRS), a cytokine storm, a cancer, or an inflammatory disease in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising an irradiated and pooled, mononuclear-enriched, early apoptotic cell preparation of claim 1 .
11. The method of claim 10 , wherein the immune disease is selected from the group comprising GVHD, arthritis, gout, or inflammatory bowel disease, and wherein said CRS or cytokine storm is the result of an infection.
12. The method of claim 11 , wherein said subject
(a) is suffering from a hematopoietic malignancy or sepsis,
(b) retains a graft-versus-tumor or graft-versus-leukemia (GVL) effect,
(c) is undergoing hematopoietic stem-cell transplantation (HSCT), or
(d) is undergoing solid organ transplantation.
13. The method of claim 12 , wherein the HSCT is allogeneic HSCT and said pharmaceutical composition comprises cells obtained from multiple allogeneic donors not HLA matched to said subject or to said donor.
14. The method of claims 10 , wherein the administering of the pharmaceutical composition is carried out up to 24 hours prior to said transplantation, at the same time as the transplantation, or is administered until 15 days following said transplantation.
15. The method of claim 10 , wherein said pharmaceutical composition is administered by intravenous injection.
16. A method for producing a pharmaceutical composition comprising an irradiated and pooled, mononuclear, early apoptotic cell preparation comprising pooled, individual, mononuclear cell populations in an early apoptotic state, said method comprising the following steps,
(a) obtaining individual, mononuclear-enriched cell populations of peripheral blood, wherein said populations are obtained from a single donor or multiple donors;
(b) freezing said mononuclear-enriched cell populations in a freezing medium comprising an anticoagulant;
(c) thawing said mononuclear-enriched cell populations;
(d) incubating said mononuclear-enriched cell populations in an apoptosis inducing incubation medium comprising methylprednisolone at a final concentration of about 10-100 μg/mL and an anticoagulant, wherein induction produces an early apoptotic cell population;
(e) resuspending said induced early apoptotic cell populations obtained in step (d) in an administration medium; and
(f) inactivating said early apoptotic cell populations from step (e), wherein said inactivation comprises irradiating said apoptotic cell populations from step (e), and wherein said irradiation comprises gamma irradiation or UV irradiation at about 20-60 Grey units (Gy); and
(g) pooling said cell populations, wherein said pooling occurs following any step (a) through (f);
wherein said method produces a pharmaceutical composition comprising an irradiated and pooled, mononuclear-enriched, early apoptotic cell preparation comprising pooled, individual, mononuclear cell populations in an early apoptotic state.
17. The method of claim 16 , wherein said inactivating step comprises
(a) decreasing the percent of non-quiescent non-apoptotic cells; or
(b) suppressing cellular activation of any living non-apoptotic cells; or
(c) reducing the proliferation of any living non-apoptotic cells; or
(d) suppressing or eliminating an immune response in said individual populations; or
(e) suppressing or eliminating cross-reactivity between said individual populations; or
(f) reducing or eliminating T-cell receptor activity in said individual populations;
or any combination thereof, within said irradiated and pooled, mononuclear, early apoptotic cell preparation as compared with non-inactivated cell populations, and wherein said produced pharmaceutical composition comprising said irradiated and pooled, mononuclear, early apoptotic cell preparation comprises a decreased percent of living non-apoptotic cells, a suppressed cellular activation of any living non-apoptotic cells, or a reduced proliferation of any living non-apoptotic cells, or any combination thereof compared with non-inactivated cell populations.
18. The method of claim 16 , wherein said obtaining said individual, mononuclear-enriched cell populations comprises obtaining white blood cell (WBC) fractions from multiple individual donors by leukapheresis.
19. The method of claim 18 , wherein said white blood cell (WBC) fractions comprise
(a) WBC fractions obtained from a blood bank; or
(b) at least one cell type selected from the group consisting of lymphocytes, monocytes, dendritic cells, and natural killer cells; or
(c) white blood cell (WBC) fractions collected from about 2 to 25 units of blood;
or any combination thereof.
20. The method of claim 16 , wherein said obtaining of said individual, mononuclear-enriched cell populations is not restricted by HLA matching said individual, mononuclear-enriched cell populations.
21. The method of claim 16 , wherein said incubating is for about 2-12 hours.
22. The method of claim 16 , wherein said individual, mononuclear-enriched cell populations comprise allogeneic cells from HLA-matched or HLA-unmatched sources with respect to a recipient subject.