Recombinant surfactant protein D compositions and methods of use thereof

Abstract

We describe an rSPD(n/CRD) polypeptide, fragment, homologue, variant or derivative thereof for use in a method of treatment or prophylaxis of a disease. A method of treating an individual suffering from a disease or preventing the occurrence of a disease in an individual is also described, in which the method comprises administering to the individual a therapeutically or prophylactically effective amount of an rSPD (n/CRD) polypeptide, fragment, homologue, variant or derivative thereof. Preferably, the rSPD (n/CRD) polypeptide and nucleic acid comprise SEQ ID NO: 1 and SEQ ID NO: 2, respectively.

1 . A recombinant polypeptide comprising amino acids 54-177 of SEQ ID NO: 1 or a fragment thereof that retains one or more biological activities of an rSPD(n/CRD) polypeptide.  
     
     
         2 . An isolated rSPD(n/CRD) polypeptide comprising amino acids 1-177 of SEQ ID NO: 1 or 179-355 of SEQ ID NO: 4 or a fragment thereof that retains one or more biological activities of the polypeptide of SEQ ID NO: 1 or SEQ ID NO: 4.  
     
     
         3 . The polypeptide of  claim 1  that further comprises the sequence of amino acids 25-53 of SEQ ID NO: 1, located immediately N-terminal to said amino acids 54-177 of SEQ ID NO: 1.  
     
     
         4 . The polypeptide of  claim 3  which comprises a conservative amino acid substitution relative to the sequence of amino acids 25-53 of SEQ ID NO: 1.  
     
     
         5 . The recombinant polypeptide of claims  1  or  2  which comprises a conservative amino acid substitution relative to the sequence of amino acids 54-177 of SEQ ID NO: 1.  
     
     
         6 . The polypeptide of  claim 1  that further comprises at least one G-Xaa-Yaa sequence located N-terminal to said polypeptide of  claim 1 .  
     
     
         7 . The polypeptide of  claim 6  which comprises a contiguous stretch of 8 of said G-Xaa-Yaa sequences.  
     
     
         8 . The polypeptide of  claim 7  wherein said contiguous stretch of 8 G-Xaa-Yaa sequences consists of amino acids 1-24 of SEQ ID NO: 1.  
     
     
         9 . The polypeptide of  claim 3  which further comprises at least one G-Xaa-Yaa sequence located immediately N-terminal to said amino acids 25-53 of SEQ ID NO: 1.  
     
     
         10 . The polypeptide of  claim 9  which comprises a stretch of 8 contiguous G-Xaa-Yaa sequences located immediately N-terminal to said amino acids 25-53 of SEQ ID NO: 1.  
     
     
         11 . The polypeptide of  claim 10  which comprises a conservative amino acid substitution relative to the sequence of SEQ ID NO: 1.  
     
     
         12 . A polypeptide comprising the amino acid sequence of SEQ ID NO: 1.  
     
     
         13 . A polypeptide consisting of the amino acid sequence of SEQ ID NO: 1.  
     
     
         14 . A method of treatment or prophylaxis of a disease or disorder, the method comprising administering a therapeutically or prophylactically effective amount of an rSPD(n/CRD) polypeptide to an individual in need of such treatment or prophylaxis.  
     
     
         15 . The method of  claim 14  wherein said rSPD(n/CRD) polypeptide comprises a polypeptide according to  claim 1  or  claim 5 .  
     
     
         16 . The method of  claim 14  wherein said disease or disorder comprises an inflammatory disease.  
     
     
         17 . The method of  claim 16  wherein said inflammatory disease is selected from the group consisting of allergy and inflammatory lung disease.  
     
     
         18 . The method of  claim 17  wherein said allergy is an allergy to the house dust mite ( Dermatophagoides  spp), a fungus or fungal spores.  
     
     
         19 . The method of  claim 18  wherein said fungus is  Aspergillus fumigatus.    
     
     
         20 . The method of  claim 14  wherein said disease or disorder is associated with microbial infection.  
     
     
         21 . The method of  claim 20  wherein said microbial infection is a bacterial or viral infection.  
     
     
         22 . The method of  claim 20  wherein said microbial infection is an infection of the lung.  
     
     
         23 . A method of reducing airway hyperresponsiveness, serum IgE levels or eosinophilia in an individual, the method comprising administering to the individual an rSPD(n/CRD) polypeptide.  
     
     
         24 . The method of  claim 23  wherein said rSPD(n/CRD) polypeptide comprises a polypeptide of  claim 1  or  claim 5 .  
     
     
         25 . A method of reducing alveolar macrophage number in an individual, the method comprising administering to the individual an rSPD(n/CRD) polypeptide.  
     
     
         26 . The method of  claim 25  wherein said rSPD(n/CRD) polypeptide comprises a polypeptide of  claim 1  or  claim 5 .  
     
     
         27 . The method of  claim 25  wherein said method enhances the clearance of apoptotic alveolar macrophages, or enhances the clearance of necrotic alveolar macrophages, or both.  
     
     
         28 . The method of  claim 14  wherein said method comprises surfactant replacement therapy.  
     
     
         29 . An isolated polynucleotide encoding an rSPD(n/CRD) polypeptide.  
     
     
         30 . A vector comprising a polynucleotide sequence encoding an rSPD(n/CRD) polypeptide.  
     
     
         31 . The vector of  claim 30  which comprises the sequence of SEQ ID NO: 2.  
     
     
         32 . A host cell comprising the vector of  claim 30 .  
     
     
         33 . The host cell of  claim 32  which comprises a vector which comprises the sequence of SEQ ID NO: 2.  
     
     
         34 . The polynucleotide of  claim 29 , wherein said polynucleotide comprises the sequence of nucleotides 157-534 of SEQ ID NO: 2.  
     
     
         35 . The polynucleotide of  claim 34 , wherein said polynucleotide comprises a mutation that results in a conservative amino acid substitution in the polypeptide encoded by said nucleotides 157-534 of SEQ ID NO: 2, wherein said encoded polypeptide retains one or more biological activities of an rSPD(n/CRD) polypeptide.  
     
     
         36 . The polynucleotide of  claim 29 , wherein said polynucleotide comprises the sequence of nucleotides 73-534 of SEQ ID NO: 2.  
     
     
         37 . The polynucleotide of  claim 29 , wherein said polynucleotide comprises the sequence of nucleotides 1-534 of SEQ ID NO: 2.  
     
     
         38 . The polynucleotide of  claim 36  or  37  which comprises a mutation that results in a conservative amino acid substitution in the polypeptide encoded by said nucleotides 1-534 of SEQ ID NO: 2, wherein said encoded polypeptide retains one or more biological activities of an rSPD(n/CRD) polypeptide.  
     
     
         39 . An isolated polynucleotide comprising the sequence of nucleotides 157-534 of SEQ ID NO: 2.  
     
     
         40 . The polynucleotide of  claim 39  which comprises a mutation that results in a conservative amino acid substitution in the polypeptide encoded by said nucleotides 157-534 of SEQ ID NO: 2, wherein said encoded polypeptide retains one or more biological activities of an rSPD(n/CRD) polypeptide.  
     
     
         41 . An isolated polynucleotide which comprises the sequence of nucleotides 73-534 of SEQ ID NO: 2.  
     
     
         42 . The polynucleotide of  claim 41  which comprises a mutation that results in a conservative amino acid substitution in the polypeptide encoded by said nucleotides 73-534 of SEQ ID NO: 2, wherein said encoded polypeptide retains one or more biological activities of an rSPD(n/CRD) polypeptide.  
     
     
         43 . An isolated polynucleotide which comprises the sequence of nucleotides 1-534 of SEQ ID NO: 2.  
     
     
         44 . The polynucleotide of  claim 43  which comprises a mutation that results in a conservative amino acid substitution in the polypeptide encoded by said nucleotides 1-534 of SEQ ID NO: 2, wherein said encoded polypeptide retains one or more biological activities of an rSPD(n/CRD) polypeptide.  
     
     
         45 . A composition comprising a recombinant polypeptide of  claim 1  or  claim 5  and a pharmaceutically acceptable carrier.  
     
     
         46 . A composition comprising a recombinant polypeptide of  claim 1  or  claim 5 , or a polynucleotide of  claim 29 , or a vector of  claim 30  or a host cell of  claim 32  and one or more of the following components: a lung extract, a phospholipid, a fatty acid, a surfactant-associated protein, a cetyl alcohol, a long-chain repeating alcohol, tyloxapol, colfosceril palmitate (dipalmitoylphosphatidylcholine), palmitic acid, tripalmitin, poractant alfa and sodium chloride.  
     
     
         47 . The composition of  claim 46  further comprising a pharmaceutically acceptable carrier.  
     
     
         48 . The composition of  claim 46  wherein said lung extract comprises a bovine lung extract.  
     
     
         49 . The composition of  claim 48  wherein said bovine lung extract comprises a calf lung extract.  
     
     
         50 . The composition of  claim 46  wherein said phospholipid comprises a phosphatidylcholine or a disaturated phosphatidylcholine.  
     
     
         51 . The composition of  claim 46  wherein said surfactant-associated protein comprises a surfactant-associated protein B or a surfactant-associated protein C.  
     
     
         52 . A composition comprising a polynucleotide of  claim 29 , a vector of  claim 30  or a host cell of  claim 32  and a pharmaceutically acceptable carrier.  
     
     
         53 . A method of identifying a molecule which binds to an rSPD(n/CRD) polypeptide, the method comprising contacting an rSPD(n/CRD) polypeptide to a candidate molecule and detecting whether the candidate molecule binds to the rSPD(n/CRD) polypeptide.  
     
     
         54 . A method of identifying an agonist or antagonist of an rSPD(n/CRD) polypeptide, the method comprising: 
 (a) providing a cell or organism;    (b) contacting the cell or organism with an rSPD(n/CRD) polypeptide;    (c) contacting the cell with a candidate molecule; and    (d) detecting a change in a biological activity of said rSPD(n/CRD), wherein said change is indicative that said candidate molecule is an agonist or antagonist of said rSPD(n/CRD) polypeptide.    
     
     
         55 . The method of  claim 54 , in which said biological activity of said rSPD(n/CRD) is selected from the group consisting of: reduction of peripheral blood eosinophilia, reduction of serum IgE levels, reduction of serum IgE levels, reduction in airway hyperresponsiveness, reduction in alveolar macrophage number, reduction of phospholipid levels in lavage, down-regulation of Eotaxin expression, reduction in MCP-1 expression, down-regulation of MIP-1α expression, and down-regulation of MIP-2 expression.  
     
     
         56 . A method for the treatment of a condition involving the production of free DNA on the surface of cells, said method comprising administering a therapeutically effective amount of an SP-D polypeptide to an individual in need of such treatment.  
     
     
         57 . The method of  claim 56  wherein said SP-D polypeptide binds to nucleic acid via its carbohydrate-binding domain.  
     
     
         58 . A method for the treatment of a condition involving the production of free DNA on the surface of cells, said method comprising administering a therapeutically effective amount of an rSPD(n/CRD) polypeptide to an individual in need of such treatment.  
     
     
         59 . A method for the treatment of a condition involving the accumulation of free nucleic acid in the lung, said method comprising administering a therapeutically effective amount of an SP-D polypeptide to an individual in need of such treatment.  
     
     
         60 . A method for the treatment of a condition involving the accumulation of free nucleic acid in the lung, said method comprising administering a therapeutically effective amount of an rSPD(n/CRD) polypeptide to an individual in need of such treatment.  
     
     
         61 . A method for the treatment of cystic fibrosis, said method comprising administering a therapeutically effective amount of an SP-D polypeptide to an individual in need of such treatment.  
     
     
         62 . A method for the treatment of cyctic fibrosis, said method comprising administering a therapeutically effective amount of an rSPD(n/CRD) polypeptide to an individual in need of such treatment.  
     
     
         63 . A method for the clearance of necrotic and apoptotic cells, said method comprising administering a therapeutically effective amount of an SP-D polypeptide to an individual in need of such clearance.  
     
     
         64 . A method for the clearance of necrotic and apoptotic cells, said method comprising administering a therapeutically effective amount of an rSPD(n/CRD) polypeptide to an individual in need of such clearance.  
     
     
         65 . The method of  claim 63  or  claim 64  wherein said necrotic or apoptotic cells are present in the lung.  
     
     
         66 . A method for the reduction of autoantibody production in autoimmune disease, the method comprising administering a therapeutically effective amount of an SP-D polypeptide to an individual in need of such reduction.  
     
     
         67 . A method for the reduction of autoantibody production in autoimmune disease, the method comprising administering a therapeutically effective amount of an rSPD(n/CRD) polypeptide to an individual in need of such reduction.  
     
     
         68 . A method for treating an autoimmune disease comprising administering to a patient in need of such treatment a pharmaceutically effective amount of an SP-D polypeptide.  
     
     
         69 . A method for treating an autoimmune disease comprising administering to a patient in need of such treatment a pharmaceutically effective amount of an rSPD(n/CRD) polypeptide.