Compositions and Methods of Use for Mgd-Csf in Disease Treatment

Abstract

Disclosed is a newly identified secreted molecule, identified herein as “monocyte, granulocyte, and dendritic cell colony stimulating factor” (MGD-CSF), the polypeptide sequence, and polynucleotides encoding the polypeptide sequence. Also provided is a procedure for producing the polypeptide by recombinant techniques employing, for example, vectors and host cells. Additionally, procedures are described to modify the disclosed novel molecules of the invention to prepare fusion molecules. Also disclosed are methods for using the polypeptides and active fragments thereof for treatment of a variety of diseases, including, for example, cancer, autoimmune and inflammatory diseases, infectious diseases, and recurrent pregnancy loss.

1 . An isolated nucleic acid molecule comprising a first polynucleotide that comprises a first nucleotide sequence chosen from 
 (a) SEQ. ID. NOS.:1, 2, 3, and 5;    (b) a first polynucleotide encoding a first polypeptide comprising a first amino acid sequence chosen from SEQ. ID. NOS.:7, 8, 9, and 11;    (c) a polynucleotide comprising a nucleotide sequence that is complementary to the first nucleotide sequence of (a);    (d) an isolated nucleic acid molecule that specifically hybridizes under stringent conditions to SEQ. ID. NOS. 1, 2, or 3; and    (e) a biologically active fragment of (a), (b), (c) or (d),    wherein the nucleic acid molecule encodes a polypeptide that can stimulate the proliferation and differentiation of granulocytes, monocytes, and dendritic cells.    
     
     
         2 . The nucleic acid molecule of  claim 1 , wherein the biologically active polypeptide fragment comprises at least six contiguous amino acid residues chosen from SEQ. ID. NOS.: 7, 8, 9, and 11, and wherein at least two of the contiguous six amino acid residues are leucine and arginine residues at amino acid residue positions 80 and 81, respectively.  
     
     
         3 - 14 . (canceled)  
     
     
         15 . An isolated polypeptide comprising a first amino acid sequence chosen from SEQ. ID. NOS.:7, 8, 9, and 11; a sequence chosen from a sequence encoded by SEQ. ID. NOS.:1, 2, 3, and 5; and a biologically active fragment of any of these.  
     
     
         16 . The polypeptide of  claim 15 , wherein the biologically active fragment comprises at least six contiguous amino acid residues chosen from SEQ. ID. NOS.:7, 8, 9, and 11, wherein at least two of the contiguous six amino acid residues are leucine and arginine at amino acid residue 80 and 81 of SEQ. ID. NO.:5.  
     
     
         17 - 27 . (canceled)  
     
     
         28 . A vector comprising the nucleic acid molecule of  claim 1  and a promoter that regulates the expression of the nucleic acid molecule.  
     
     
         29 - 31 . (canceled)  
     
     
         32 . A recombinant host cell comprising a cell and the nucleic acid of  claim 1 , the polypeptide of  claim 15 , and/or the vector of  claim 28 .  
     
     
         33 - 41 . (canceled)  
     
     
         42 . A nucleic acid composition comprising the nucleic acid molecule of  claim 1 , and a carrier.  
     
     
         43 . A polypeptide composition comprising the polypeptide molecule of  claim 15  and a carrier.  
     
     
         44 . A vector composition comprising the vector of  claim 28  and a carrier.  
     
     
         45 . A host cell composition comprising the host cell of  claim 32  and a carrier.  
     
     
         46 - 49 . (canceled)  
     
     
         50 . A method of producing a recombinant host cell comprising: 
 (a) providing a vector that comprises the nucleic acid molecule of  claim 1;  and    (b) allowing a cell to come into contact with the vector to form a recombinant host cell transfected with the nucleic acid molecule.    
     
     
         51 . A method of producing a polypeptide comprising: 
 (a) providing the nucleic acid molecule of  claim 1;  and    (b) expressing the nucleic acid molecule in an expression system to produce the polypeptide.    
     
     
         52 - 58 . (canceled)  
     
     
         59 . A diagnostic kit comprising a composition that comprises a nucleic acid molecule of  claim 1 , a reporter for detecting the nucleic acid molecule or its complement, and a vehicle.  
     
     
         60 . A diagnostic kit comprising an antibody that specifically binds to a polypeptide of  claim 15 , and a carrier.  
     
     
         61 . A diagnostic kit comprising a polypeptide of  claim 15  and a carrier.  
     
     
         62 . A method of determining the presence of an antibody specific to a polypeptide of  claim 15  in a patient sample comprising: 
 (a) providing a composition comprising a polypeptide of  claim 15;     (b) allowing the polypeptide to interact with the sample; and    (c) determining whether interaction has occurred between the polypeptide and antibody in the sample, if present.    
     
     
         63 . An isolated antibody that specifically binds to and/or interferes with the activity of an antigen that comprises at least six contiguous amino acid residues chosen from SEQ. ID. NOS.:7-12.  
     
     
         64 . The antibody of  claim 63 , wherein the contiguous amino acid residues comprise the consecutive amino acid residues leu-arg at amino acid positions 80 and 81 of SEQ. ID. NO.:7 or the consecutive amino acid residues leu-gln-arg of SEQ. ID. NO.:12.  
     
     
         65 - 66 . (canceled)  
     
     
         67 . The polypeptide of  claim 15  or a polypeptide produced by the method of  claim 51 , wherein the polypeptide further comprises at least one fusion partner.  
     
     
         68 - 71 . (canceled)  
     
     
         72 . A method of screening for an agent that modulates activity of the polypeptide of  claim 15  comprising: 
 (a) providing a test system in which a polypeptide of  claim 15  affects biological activity; and    (b) screening multiple agents for an effect on the activity of the polypeptide of  claim 15  on the test system.    
     
     
         73 - 74 . (canceled)  
     
     
         75 . A method of stimulating the proliferation of and/or the activation of immune cells comprising: 
 (a) providing a composition comprising a substantially pure polypeptide chosen from any of SEQ. ID. NOS.:7-12, and active fragments thereof; and    (b) contacting one or more immune cells with the polypeptide.    
     
     
         76 - 92 . (canceled)  
     
     
         93 . A method of treating or preventing cancer in a subject comprising: 
 (a) providing a composition comprising a substantially pure polypeptide chosen from SEQ. ID. NOS.:7-12 and active fragments of any of these; and    (b) administering the composition to the subject.    
     
     
         94 - 96 . (canceled)  
     
     
         97 . A method of treating or preventing an infection in a subject comprising: 
 (a) providing a composition comprising a substantially pure polypeptide chosen from SEQ. ID. NOS.:7-12 and active fragments of any of these; and    (b) administering the composition to the subject.    
     
     
         98 - 100 . (canceled)  
     
     
         101 . A method for modulating an immune response in a subject, comprising: 
 (a) providing a modulator of a polypeptide chosen from SEQ. ID. NOS.:7-12 and active fragments of any of these; and    (b) administering the modulator to the subject.    
     
     
         102 - 129 . (canceled)  
     
     
         130 . A method of increasing a population of hematopoietic stem cells comprising: 
 (a) providing a composition comprising a substantially pure polypeptide chosen from SEQ. ID. NOS.:7-12 and active fragments of any of these; and    (b) contacting the population of hematopoietic stem cells with the polypeptide.    
     
     
         131 . A method of providing cytoprotection to a population of cells comprising: 
 (a) providing a composition comprising a substantially pure polypeptide chosen from SEQ. ID. NOS.:7-12 and active fragments of any of these; and    (b) contacting the population of cells with the polypeptide.    
     
     
         132 - 139 . (canceled)