CD14+ Cell Compositions and Methods of Using Same
The present invention provides CD 14 + cell compositions and methods of using same in treating disorders, such as inflammatory disorders, such as atherosclerosis and cardiovascular disease.
1 . A composition comprising a population of cells of hematopoietic lineage, wherein the composition contains CD14 + macrophages, and wherein when the cells are contacted with a pro-inflammatory stimulus produce inflammatory cytokines such that the anti-inflammatory cytokine: pro-inflammatory cytokine ratio produced is at least 2:1.
2 . The composition of claim 1 , wherein the composition further comprises CD14 + monocytes.
3 . The composition of claim 1 , wherein the ratio is at least 5:1, 10:1, 25:1, 50:1 or 100:1.
4 . The composition of claim 1 , wherein the cells are derived from bone marrow, peripheral blood, umbilical cord blood, fetal liver, human embryonic stem cells (huES), induce pluripotent stem cells (iPS) or parthenogenetic cells.
5 . The composition of claim 1 , wherein the composition has one or more of the following characteristics:
a) the viability of the cells is at least 75%;
b) contains less than 2 μg/ml serum albumin;
c) substantially free of horse serum or
d) substantially free of mycoplasm, endotoxin and microbial contamination.
6 . The composition of claim 1 , wherein the cells are in a pharmaceutical-grade electrolyte solution suitable for human administration.
7 . The composition of claim 1 , wherein the total number of cells is 40 to 200 million.
8 . The composition of claim 1 , wherein the cells are in a volume less than 15 mLs.
9 . The composition of claim 1 , wherein the cells produce at least 100 pg per 2×10 6 cells of one or more anti-inflammatory cytokines.
10 . The composition of claim 1 , where in the anti-inflammatory cytokine is IL-10 or ILRa.
11 . The composition of claim 1 , wherein the pro-inflammatory stimulus is lipopolysaccharide (LPS).
12 . The composition of claim 1 , wherein at least 5% of the CD14 + macrophages are autofluorescent.
13 . The composition of claim 1 , wherein said composition is an in-vitro expanded cell population.
14 . The composition of claim 2 , wherein the CD14 + monocytes are expanded in vitro.
15 . The composition of claim 14 , wherein the CD14 + monocytes differentiate into CD14 + macrophages in vitro.
16 . The composition of claim 1 , wherein the CD14 + macrophages are derived from CD34 + hematopoietic progenitor cells that have been differentiated in vitro.
17 . The composition of claim 16 , wherein the CD34+ hematopoietic progenitor are myeloid cells.
18 . The composition of claim 17 , wherein the myeloid cells are myeolomonocytes.
19 . The composition of claim 1 , wherein the cells are isolated from an in-vitro expanded cell culture.
20 . The composition of claim 19 , wherein in-vitro expanded cell culture is derived from mononuclear cells.
21 . The composition of claim 19 , wherein in-vitro expanded cell culture comprises a mixed population of cells of hematopoietic, mesenchymal and endothelial linage.
22 . The composition of claim 19 , wherein in-vitro expanded cell culture comprises a mixed population of cells of hematopoietic and mesenchymal linage.
23 . The composition of claim 19 , wherein in-vitro expanded cell culture comprises a population of hematopoietic cells.
24 . The composition of claim 21 or 22 , wherein the mixed population of cells are about 5-75% viable CD90 + cells with the remaining cells in the composition being CD45 + .
25 . The composition of claim 23 , wherein the hematopoietic cells are CD45 + .
26 . The composition of claim 1 , wherein at least 5% of the CD14 + macrophages are CD66b-negative, CD18 + , CD33 + , CD11b + , CD11c + , CD91-negative, CD141 + , HLA-DR-negative, CD209-negative, CD16-negative, and/or CD1c-negative.
27 . The composition of claim 26 , wherein at least 10% of the CD14+ macrophages are CD66b-negative, CD18+, CD33+, CD11b+, CD11c+, CD91-negative, CD141+, HLA-DR-negative, CD209-negative, CD16-negative, and/or CD1c-negative.
28 . The composition of claim 27 , wherein at least 15% of the CD14+ macrophages are CD66b-negative, CD18+, CD33+, CD11b+, CD91-negative, CD141+, HLA-DR-negative, CD209-negative, CD16-negative, and/or CD1c-negative.
29 . The composition of claim 1 , wherein at least 5% of the CD14 + macrophages express PPARγ, CD206, CD163, CD204, SR-B1, MERTK, and/or TGFβ.
30 . The composition of claim 29 , wherein at least 10% of the CD14 + macrophages express PPARγ, CD206, CD163, CD204, SR-B1, MERTK, and/or TGFβ.
31 . The composition of claim 30 , wherein at least 15% of the CD14 + macrophages express PPARγ, CD206, CD163, CD204, SR-B1, MERTK, and/or TGFβ.
32 . The composition of claim 1 , wherein the CD14 + macrophages express a higher level of PPARγ, CD206, CD163, CD204, SR-B1, MERTK, and/or TGFβ compared to M1 macrophages.
33 . The composition of claim 32 , wherein the CD14 + macrophages express an at least two-fold higher level of PPARγ, CD206, CD 163, CD204, SR-B 1, MERTK, and/or TGF compared to M1 macrophages.
34 . The composition of claim 1 , wherein the CD14 + macrophages express a lower level of CCR7, IL-1B, and/or TNFα compared to M1 macrophages.
35 . The composition of claim 34 , wherein the CD14 + macrophages express an at least two-fold lower level of CCR7, IL-1B, and/or TNFα compared to M1 macrophages.
36 . The composition of claim 1 , wherein the CD14 + macrophages are exposed to a pro-inflammatory stimulus, and wherein the expression level of a pro-inflammatory cytokine in the CD14 + macrophages after the exposure is 100% or less of the expression level of the pro-inflammatory cytokine prior to the exposure.
37 . The composition of claim 36 , wherein the pro-inflammatory stimulus comprises a pathogen, lipopolysaccharide, interferon-gamma, lipoxin, a leukotriene, an endotoxin, or debris from a dead cell.
38 . The composition of claim 36 , wherein the pro-inflammatory cytokine comprises TNFβ, IL-1A, IL-1B, or IL-12.
39 . A method of modulating cholesterol efflux in vascular tissue of a subject comprising administering to a subject in need thereof the composition of claim 1 or a composition comprising ixmyelocel-T.
40 . A method of decreasing atherosclerotic lesions in a subject comprising administering to a subject in need thereof the composition of claim 1 or a composition comprising ixmyelocel-T.
41 . A method of treating atherosclerosis comprising administering to a subject in need thereof the composition of claim 1 or a composition comprising ixmyelocel-T.
42 . A method of decreasing oxidative stress of a tissue comprising contacting the tissue with composition of claim 1 or a composition comprising ixmyelocel-T.
43 . The method of claim 42 , wherein the tissue is endothelium.
44 . A method of increasing plasma nitrate levels and/or decreasing plasma lipid peroxidation in a subject comprising administering to a subject in need thereof the composition of claim 1 or a composition comprising ixmyelocel-T.
45 . A method of increasing the expression of endothelial nitric oxide synthase (eNOS) and/or nitric oxide production (NO) in a cell comprising contacting the cell with composition of claim 1 or a composition comprising ixmyelocel-T.
46 . A method of tissue regeneration or repair comprising administering to a patient in need thereof the composition of claim 1 .
47 . A method of treating ischemic disorders comprising administering to a patient in need thereof the composition of claim 1 .
48 . A method of inducing angiogenesis in a tissue comprising administering to a patient in need thereof the composition of claim 1 .