US2005130303A1PendingUtilityA1

Stimulation of hematopoiesis by ex vivo activated immune cells

52
Priority: May 29, 2002Filed: Nov 9, 2004Published: Jun 16, 2005
Est. expiryMay 29, 2022(expired)· nominal 20-yr term from priority
Inventors:Demao Yang
A61P 7/00C12N 2501/22C12N 2500/14A61P 7/06C12N 2501/23A61K 35/14A61K 40/416A61K 40/22A61K 40/10A61K 2239/31C12N 5/0634
52
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Claims

Abstract

A protocol of activating and administering human blood cells so that bone marrow histology and/or blood cell counts of patients suffering from a plastic anemia approach normal. The protocol includes culturing the blood cells in the presence of a cytokine and an ionophore. It is emphasized that this abstract is provided to comply with the rules requiring an abstract that will allow a searcher or other reader to quickly ascertain the subject matter of the technical disclosure. It is submitted with the understanding that it will not be used to interpret or limit the scope or meaning of the claims. 37 C.F.R. § 1.72(b).

Claims

exact text as granted — not AI-modified
1 . A method of culturing blood cells, comprising culturing the blood cells in the presence of an ionophore and between about 10 and about 50,000 IU/ml of a cytokine.  
     
     
         2 . The method of  claim 1 , in which the blood cells are cultured in the presence of effective amounts of the cytokine and the ionophore.  
     
     
         3 . The method of  claim 1 , in which the blood cells are cultured in a medium comprising mammalian serum.  
     
     
         4 . The method of  claim 1 , in which the blood cells are cultured for a period between about 10 and 90 hours.  
     
     
         5 . The method of  claim 1 , in which the blood cells are cultured at a temperature between about 30 and 42 degrees C.  
     
     
         6 . The method of  claim 1 , in which the blood cells are cultured in the presence of interleukin-2 and granulocyte macrophage-colony stimulating factor.  
     
     
         7 . The method of  claim 1 , in which the blood cells are allogeneic to a patient.  
     
     
         8 . The method of  claim 1 , in which the blood cells are from an immunologically acceptable owner.  
     
     
         9 . The method of  claim 1 , in which the blood cells are autologous to a patient.  
     
     
         10 . The method of  claim 1 , in which the blood cells are cultured in the presence of A23187.  
     
     
         11 . The method of  claim 1 , in which the blood cells are at least partially separated from sera before being cultured.  
     
     
         12 . The method of  claim 1 , in which the blood cells are at least partially separated from sera by being centrifuged before being cultured.  
     
     
         13 . The method of  claim 1 , in which the blood cells are at least partially separated from blood by Ficoll-Hypaque centrifugation of the blood.  
     
     
         14 . The method of  claim 1 , further comprising culturing with a culture medium having at least one amino acid.  
     
     
         15 . The method of  claim 1 , wherein the ionophore is present in a concentration of between about 1 and about 10,000 ng/ml.  
     
     
         16 . The method of  claim 1 , wherein the ionophore is present in a concentration of between about 10 and about 500 ng/ml.  
     
     
         17 . The method of  claim 1 , wherein the cytokine is present in a concentration of between about 100 and about 5,000 IU/ml.  
     
     
         18 . A quantity of activated blood cells effective to treat blood deficiencies following injection into a patient.  
     
     
         19 . The quantity of blood cells of  claim 18 , in which the quantity of blood cells is cultured in the presence of a cytokine and an ionophore.  
     
     
         20 . The quantity of blood cells of  claim 19 , in which the quantity of blood cells is cultured in the presence of between about 10 and about 50,000 IU/ml of the cytokine.  
     
     
         21 . The quantity of blood cells of  claim 19 , in which the ionophore is present in a concentration of between about 1 and about 10,000 ng/ml.  
     
     
         22 . The quantity of blood cells of  claim 19 , in which the quantity of blood cells is cultured in the presence of effective concentrations of the cytokine and ionophore.  
     
     
         23 . The quantity of blood cells of  claim 18 , in which a therapeutically effective amount of the cultured blood cells elevates peripheral blood cell populations in the patients when administered to the patients.  
     
     
         24 . The quantity of blood cells of  claim 18 , in which the quantity of blood cells is cultured in the presence of an interleukin and an ionophore.  
     
     
         25 . The quantity of blood cells of  claim 18 , in which the quantity of blood cells is cultured in the presence of interleukin-2, macrophage-colony stimulating factor, and a calcium ionophore.  
     
     
         26 . The quantity of blood cells of  claim 18 , in which the quantity of blood cells is cultured in the presence of interleukin-2, granulocyte macrophage-colony stimulating factor, and A23187.  
     
     
         27 . A blood cell culture, comprising blood cells, an ionophore, and a concentration of between about 10 and about 50,000 IU/ml of a cytokine.  
     
     
         28 . The blood cell culture of  claim 27 , wherein the blood cells are present in a therapeutically effective amount.  
     
     
         29 . The blood cell culture of  claim 27 , further comprising mammalian serum.  
     
     
         30 . The blood cell culture of  claim 27 , wherein the culture has a temperature between about 30 and about 42 degrees C.  
     
     
         31 . The blood cell culture of  claim 27 , wherein the medium further comprises interleukin-2 and granulocyte macrophage-colony stimulating factor.  
     
     
         32 . The blood cell culture of  claim 27 , wherein the blood cells are allogeneic relative to a patient.  
     
     
         33 . The blood cell culture of  claim 27 , wherein the blood cells are immunologically acceptable relative to a patient.  
     
     
         34 . The blood cell culture of  claim 27 , wherein the blood cells are autologous relative to a patient.  
     
     
         35 . The blood cell culture of  claim 27 , wherein the ionophore comprises A23187.  
     
     
         36 . The blood cell culture of  claim 27 , wherein the blood cells are at least partially free of sera of the source blood of the blood cells.  
     
     
         37 . The blood cell culture of  claim 27 , wherein the blood cells are of a type derived from centrifuged sera.  
     
     
         38 . The blood cell culture of  claim 27 , wherein the blood cells are of a type derived from Ficoll-Hypaque centrifugation of blood.  
     
     
         39 . The blood cell culture of  claim 27 , further comprising a culture medium having at least one amino acid.  
     
     
         40 . The blood cell culture of  claim 27 , wherein the ionophore is present in a concentration of between about 1 and about 10,000 ng/ml.  
     
     
         41 . The blood cell culture of  claim 27 , wherein the ionophore is present in a concentration of between about 10 and about 500 ng/ml.  
     
     
         42 . The blood cell culture of  claim 27 , wherein the cytokine is present in a concentration of between about 100 and about 5,000 IU/ml.

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