Therapeutic use for alpha1 proteinase inhibitor in hematopoiesis

Abstract

A previously unrecognized fundamental property of α 1 PI is to regulate the phenotypic composition of circulating and tissue-associated cells derived from hematopoietic stem cells. The present invention comprises screening for various unmodified and modified α 1 PI's which are useful in the treatment of abnormalities in the number of cells of myeloid or lymphoid lineage that are associated with HIV-1 infection, microbial infection, leukemia, solid tumor cancers, atherosclerosis, autoimmunity, stem cell transplantation, organ transplantation, and other diseases affected by cells of the immune system. The interaction of α 1 PI with its receptors, HLE CS and LRP, influences the level of cells of different lineages. Genetic and proteolytic modification of α 1 PI is used to target these receptors to increase or decrease specific cell populations, as needed, in the various disease states.

1 . A method for identifying a modified α1proteinase inhibitor as suitable for use in treating a disease, disorder or condition in a subject, comprising: 
 (a) producing the modified α1proteinase inhibitor; and    (b) measuring a biological activity of the modified α1proteinase inhibitor in a biological assay for predicting effectiveness in treating the disease, disorder or condition in the subject,    wherein the modified α1proteinase inhibitor is identified as suitable for treating the disease, disorder or condition from a change in the biological activity relative to a control activity measured for a wild-type α1proteinase inhibitor.    
   
   
       2 . The method of  claim 1 , wherein the modified α1proteinase inhibitor is produced by site-directed mutagenesis, proteolysis, or both.  
   
   
       3 . The method of  claim 1 , wherein the disease, disorder or condition is selected from the group consisting of HIV-1 infection, bacterial infection, leukemia, a solid tumor, atherosclerosis, an autoimmune disease, organ transplantation, and stem cell transplantation.  
   
   
       4 . The method of  claim 1 , wherein the biological assay is selected from the group consisting of an elastase inhibition assay, a receptor co-capping assay, a cell motility assay, a lymphoid-committed progenitor cell mobilization assay, an HIV-1 gp120 antibody cross-reactivity assay, and an HIV-1 infectivity facilitation assay.  
   
   
       5 . The method of  claim 1 , wherein the subject is a human or a non-human animal.  
   
   
       6 . The method of  claim 2 , wherein proteolysis comprises contacting a wild-type or a recombinant α1proteinase inhibitor with a protease selected from the group consisting of elastase, stromelysin-3, matrix metalloproteinase, collagenase, gelatinase, pepsin, plasmin, urokinase, chymotrypsin, thrombin, CD26, complement component C1, and complement component C3.  
   
   
       7 . The method of  claim 2 , wherein site-directed mutagenesis comprises changing a wild-type amino acid selected from the group consisting of residues 370-374 and 385 to a non-wild-type residue.  
   
   
       8 . The method of  claim 7 , wherein at least one amino acid selected from the group consisting of residues 370-374 and 385 is changed from wild-type to glycine, threonine, or a hydrophobic amino acid.  
   
   
       9 . The method of  claim 8 , wherein the hydrophobic amino acid is selected from the group consisting of isoleucine, leucine, phenylalanine, tyrosine and valine.  
   
   
       10 . A modified human α1proteinase inhibitor comprising a change in a wild-type amino acid residue selected from the group consisting of residues 370-374 and 385.  
   
   
       11 . The modified human α1proteinase inhibitor of  claim 10 , wherein the modification further comprises proteolysis.  
   
   
       12 . The modified human α1proteinase inhibitor of  claim 10 , wherein the change is to glycine, threonine, or a hydrophobic amino acid.  
   
   
       13 . The modified human α1proteinase inhibitor of  claim 12 , wherein the hydrophobic amino acid is selected from the group consisting of isoleucine, leucine, phenylalanine, tyrosine and valine.  
   
   
       14 . The modified human α1proteinase inhibitor of  claim 10 , which comprises at least two changes in wild-type amino acid residues selected from the group consisting of residues 370-374 and 385.  
   
   
       15 . The modified human α1proteinase inhibitor of  claim 14 , wherein methionine at position 385 is changed to a non-methionine amino acid.  
   
   
       16 . The modified human α1proteinase inhibitor of  claim 15 , wherein the non-methionine amino acid is selected from the group consisting of glycine, isoleucine, leucine, phenylalanine, threonine, and valine.  
   
   
       17 . The modified human α1proteinase inhibitor of  claim 16  which is capable of a reduced binding activity in an HIV-1 gp120 antibody cross-reactivity assay, relative to a wild-type α1proteinase inhibitor.  
   
   
       18 . The modified human α1proteinase inhibitor of  claim 10  comprising the following three amino acid substitutions: Phe372Gly; Leu373Gly; and Met 385Val.  
   
   
       19 . The modified human α1proteinase inhibitor of  claim 10  consisting of the following three amino acid substitutions: Phe372Gly; Leu373Gly; and Met 385Val.  
   
   
       20 . A method of treating a disease, disorder or condition in a subject in need of said treatment, comprising administering an amount effective to treat said disease disorder or condition of a modified α1proteinase inhibitor to the subject.  
   
   
       21 . The method of  claim 20 , wherein the modified α1proteinase inhibitor is produced by site-directed mutagenesis, proteolysis, or both.  
   
   
       22 . The method of  claim 20 , wherein the disease, disorder or condition is selected from the group consisting of HIV-1 infection, bacterial infection, leukemia, a solid tumor, atherosclerosis, an autoimmune disease, organ transplantation, and stem cell transplantation.  
   
   
       23 . The method of  claim 20 , wherein the subject is a human or a non-human animal.  
   
   
       24 . The method of  claim 20 , wherein the modified α1proteinase inhibitor comprising a change in a wild-type amino acid residue selected from the group consisting of residues 370-374 and 385.  
   
   
       25 . The method of  claim 24 , wherein wild-type methionine at position 385 is changed to a non-methionine amino acid, and wherein the non-methionine amino acid is selected from the group consisting of glycine, isoleucine, leucine, phenylalanine, threonine, and valine.  
   
   
       26 . The method of  claim 25 , wherein the modified α1proteinase inhibitor is capable of a reduced binding activity in an HIV-1 gp120 antibody cross-reactivity assay, relative to a wild-type α1proteinase inhibitor.  
   
   
       27 . The method of  claim 25 , wherein the modified α1proteinase inhibitor comprises the following three amino acid substitutions: Phe372Gly; Leu373Gly; and Met 385Val.  
   
   
       28 . The method of  claim 25 , wherein the modified α1proteinase inhibitor consists of the following three amino acid substitutions: Phe372Gly; Leu373Gly; and Met 385Val.  
   
   
       29 . The method of  claim 27 , further comprising administration of antiretroviral therapy.  
   
   
       30 . The method of  claim 20 , wherein the effective amount of modified α1proteinase inhibitor is a dose equivalent to about 42 mg/kg of active wild-type α1proteinase inhibitor.  
   
   
       31 . The method of  claim 3 , wherein stem cell transplantation is autologous.  
   
   
       32 . A method of treating a disease, disorder or condition in a subject in need of said treatment, comprising administering an amount effective to treat said disease-disorder or condition of an active α1proteinase inhibitor to the subject, wherein the disease, disorder or condition is selected from the group consisting of HIV-1 infection, bacterial infection, leukemia, a solid tumor, atherosclerosis, an autoimmune disease, organ transplantation, and stem cell transplantation.  
   
   
       33 . The method of  claim 32 , wherein stem cell transplantation is autologous.  
   
   
       34 . A method of treating a disease, disorder or condition in a subject in need of said treatment, comprising administering an amount effective to treat said disease disorder or condition of an active α1proteinase inhibitor to the subject, wherein the subject is characterized as having an abnormal number of lymphocytes, monocytes, or dendritic cells.  
   
   
       35 . A method of treating a disease, disorder or condition in a subject in need of said treatment, comprising administering an amount effective to treat said disease disorder or condition of an inactive α1proteinase inhibitor to the subject, wherein the disease, disorder or condition is selected from the group consisting of bacterial infection, neutropenia and immunosuppression.  
   
   
       36 . A method of treating a disease, disorder or condition in a subject in need of said treatment, comprising administering an amount effective to treat said disease disorder or condition of an inactive α1proteinase inhibitor to the subject, wherein the subject is characterized as having an abnormal number of granulocytes, monocytes, dendritic cells, eosinophils, or basophils.  
   
   
       37 . The method of  claim 32 , wherein the subject is a human or a non-human animal.  
   
   
       38 . A method for treating a subject suffering from abnormalities in the number of cells of myeloid or lymphoid lineage that are associated with HIV-1 infection, microbial infection, leukemia, solid tumor cancers, atherosclerosis, autoimmunity, stem cell transplantation or organ transplantation comprising administering to a subject in need of such treatment an amount effective to treat said subject of an α1proteinase inhibitor.  
   
   
       39 . The method of  claim 38  wherein said subject is a human or a non-human or animal.  
   
   
       40 . The method of  claim 38 , wherein said stem cell transplantation is autologous.  
   
   
       41 . The method of  claim 28 , further comprising administration of antiretroviral therapy.  
   
   
       42 . The method of  claim 33 , wherein the subject is a human or a non-human animal.  
   
   
       43 . The method of  claim 34 , wherein the subject is a human or a non-human animal.  
   
   
       44 . The method of  claim 35 , wherein the subject is a human or a non-human animal.  
   
   
       45 . The method of  claim 36 , wherein the subject is a human or a non-human animal.