Treatment of immunosuppression-related disorders

Abstract

Disclosed are methods of using blastomere-like stem cells to decrease a level of myeloid-derived suppressor cells (MDSCs). A method for decreasing a level of myeloid-derived suppressor cells (MDSCs) in a human subject, including: injectionally, orally or transdermally administering to the human subject an amount of human blastomere-like stem cells, autologous to the human subject, at 1×10.sup.8 to 1×10.sup.11 cells each time once every two weeks, for example.

Claims

1. A method for decreasing a level of myeloid-derived suppressor cells (MDSCs) in a human subject, comprising: administering intravenously to said human subject an amount of human CD10.sup.−CD66e.sup.+ blastomere-like stem cells autologous to said human subject so as to decrease said level of MDSCs in the blood of said human subject as compared to the level of MDSCs in the blood of said human subject without the treatment of human CD10.sup.−CD66e.sup.+ blastomere-like stem cells, wherein said human blastomere-like stem cells are isolated from the blood of said human subject.

2. The method of claim 1 further comprising said administering to said human subject said amount of human blastomere-like stem cells at 1×10.sup.8 to 1×10.sup.11 cells each time.

3. The method of claim 1 further comprising said administering to said human subject said amount of human blastomere-like stem cells at 5×10.sup.8 to 5×10.sup.10 cells each time.

4. The method of claim 1 further comprising said administering to said human subject said amount of human blastomere-like stem cells at 1×10.sup.9 to 1×10.sup.10 cells each time.

5. The method of claim 1 further comprising said administering to said human subject said amount of human blastomere-like stem cells once every two weeks.

6. The method of claim 1, wherein said human subject has a lung cancer.

7. The method of claim 1, wherein said human subject has a cellular proliferative disorder.

8. The method of claim 1, wherein the level of MDSCs is the percentage of MDSCs among the total peripheral blood mononuclear cells in a blood sample obtained from the subject.

Description

EXAMPLE 1

Isolation of BLSCs

(1) Methods for activation, purification, and expansion of BLSCs have been described in WO/2007/100845. In this example, BLSCs were purified from the blood of human subjects using two methods, a plasma fraction method and a hemolysis method.

(2) Briefly, for the plasma fraction method, a whole blood sample (1 ml) was prepared from a first human subject using a standard method. The sample was then stored at 4° C. for about 7-9 days and BLSCs were enriched from the sample in the manner described in WO/2007/100845.

(3) The hemolysis method was used to obtain a hemolysis fraction in the manner described in WO/2007/100845. Briefly, about 1 ml of whole blood is obtained from a second human subject and stored at about 4° C. in the presence of EDTA or other Ca.sup.2+ complexing agents for about 9 days in a transport medium (e.g., Moraga medium with catalog number MBC-HUB-MED-100-A004, Moraga Biotechnology Corporation, Los Angeles, Calif.). After 9 days, the red cells in the whole blood sample were lysed using about 50 ml of a sterile hemolysis solution (e.g., MBC-ASB-REBG-900A-001). After centrifugation (e.g., at 1800×g, 10 minutes) to remove debris and lysed cells, the cell pellet was re-suspended in 2 ml of a Moraga sterile reconstitution solution (MBC-ASB-REBG-900A-002).

(4) The above-described two cell populations were analyzed by flow cytometry using FITC-labeled anti-CD10 antibody, PE-labeled anti-CD66e antibody, APC-labeled anti-CD 90 antibody. The results are summarized in Table 1 below.

(5) TABLE-US-00001 TABLE 1 Percentage of Isolated Cells (%) Plasma Markers Hemolysis method fraction method CD10.sup.−CD66e.sup.+ 5.81 9.14 CD10.sup.+CD66e.sup.+ 66.67 2.99 CD10.sup.+CD66e.sup.− 3.11 1.10 CD10.sup.−CD90.sup.+ 0.62 9.80 CD10.sup.+CD90.sup.+ 13.65 2.20 CD10.sup.+CD90.sup.− 55.13 1.46

(6) As shown in Table 1, when using the hemolysis method, about 5.81%, 66.67%, and 3.11% of the isolated cells were BLSCs (CD10.sup.−CD66e.sup.+), transitional BLSCs (trBLSCs, CD10.sup.+CD66e.sup.+), and epiblast-like stem cells (ELSCs, CD10.sup.+CD66e.sup.−), respectively. About 0.62%, 13.65%, and 55.13% of the cells were CD10.sup.−CD90.sup.+, CD10.sup.1CD90.sup.1 (transitional epiblast-like stem cells, trELSCs), and CD10.sup.1CD90.sup.−, respectively. BLSCs were further enriched based on their markers (CD10.sup.−CD66e.sup.+). This method yielded about 200×10.sup.6 BLSCs/ml blood.

(7) When using the plasma faction methods, about 9.14%, 2.99%, and 1.10% of the isolated cells were BLSCs, transitional BLSCs, and ELSCs, respectively. About 9.8%, 2.2%, and 1.46% of the cells were CD10.sup.−CD90.sup.1, CD10.sup.1CD90 (trELSCs), CD10.sup.1CD90.sup.−, respectively. This method yielded about 239×10.sup.6 BLSCs/ml plasma.

(8) The BLSCs were trypan blue staining positive and generally smaller than 1 μm in size, which was different from that of platelets (CD42.sup.+ and trypan blue staining negative). Especially, unlike platelets, which lack nuclei, do not proliferate and differentiate, the BLSCs could proliferate in a medium and be maintained and expanded in an undifferentiated status in the manner described in WO/2007/100845. The BLSCs lacked contact inhibition and could form sphere-like cell aggregation, multiple confluent layers, and mesh-net structures. Aggregation of the cells led to change the cell morphology. In contrast, CD42.sup.+ cells or platelets did not form the just-mentioned structures, such as cell aggregation.

(9) The BLSCs were then tested for their differentiation capacity in the manner described in WO/2007/100845 or other methods known in the art. It was found that, upon induction under conditions known in the art, the cells differentiated into various lineages including those derived from ectoderm, mesoderm, endoderm, and spermatogonia. They could be maintained and expanded in the undifferentiated status for over 300 passages without losing the differentiation potentials. They did not form teratoma.

EXAMPLE 2

In Vivo Activity of BLSCs

(10) BLSCs were purified from a human subject according to the methods described above and administered autologously to the same subject at 1×10.sup.9 cells. At Days 0, 14, and 28 after administration, blood samples were obtained from the person. Cytometry analysis was then conducted to examine the blood levels of MDSCs and Treg. It was found that, at Days 0, 14, and 28, MDSCs (CD14.sup.−CD33.sup.+CD11b.sup.+Lin.sup.−) levels were 9.24%, 2.19%, and 0.35% of total peripheral blood mononuclear cells (PBMC), respectively. These results indicate BLSCs significantly decreased the level of MDSCs in a subject and therefore can be used to treat patients having a MDSCs-related immunodeficiency disorder.

Other Embodiments

(11) All of the features disclosed in this specification may be combined in any combination. Each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.

(12) From the above description, one skilled in the art can easily ascertain the essential characteristics of the present invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions. Thus, other embodiments are also within the scope of the following claims.