Heat shock genes and proteins from Neisseria meningitidis, Candida glabrata and Aspergillus fumigatus

Abstract

Methods and compositions comprising isolated nucleic acid molecules specific to Neisseria meningitidis, Candida glabrata and Aspergillus fumigatus heat shock proteins (Hsps), as well as vector constructs and isolated polypeptides specific to the same are provided. Such compositions and methods are useful for the diagnosis of infections by these organisms and for generating an immune response to the organisms.

1 . An isolated nucleic acid molecule encoding a  Neisseria meningitidis  Hsp70.  
     
     
         2 . An isolated nucleotide molecule selected from the group consisting of: 
 (a) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 1;    (b) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 1 from nucleotides 358-2286;    (c) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 2 from nucleotides 4-1932;    (d) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 3;    (e) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 4; and    (f) an isolated nucleic acid molecule comprising a sequence complementary to any one of the isolated nucleotide molecules set forth in (a) through (e), respectively:    
     
     
         3 . An isolated nucleic acid molecule which is a variant of, or is substantially similar to, the nucleotide molecule according to  claim 2 .  
     
     
         4 . An isolated nucleic acid molecule comprising a nucleotide sequence that is identical to a segment of contiguous nucleotide bases comprising at least 25% of any one of: SEQ. ID NO: 1 from nucleotides 358-2286; SEQ. ID NO: 2 from nucleotides 4-1932; SEQ ID NO: 3; SEQ. ID NO: 4; or to a complement thereof.  
     
     
         5 . An isolated nucleic acid molecule comprising a nucleic acid sequence that encodes a polypeptide comprising any one of the polypeptides according to FIGS. 2, 4,  6 ,  8 , or  9  or a variant Hsp70 that is at least 95% homologous to a polypeptide according to any one of FIGS. 2, 4,  6 ,  8 , or  9 , wherein percent homology is determined according to an algorithm incorporated in a protein database search program used in BLAST™ or DNA Star Megalign™.  
     
     
         6 . The isolated nucleic acid molecule according to  claim 5  wherein the encoded polypeptide is able to selectively bind an antibody specific for a  Neisseria meningitidis  Hsp70.  
     
     
         7 . An isolated nucleic acid molecule encoding at least 8 contiguous amino acids of a  Neisseria meningitis  Hsp70 polypeptide selected from amino acid residues of FIG. 6, wherein the encoded  Neisseria meningitidis  Hsp70 polypeptide is able to bind to a major histocompatibility complex.  
     
     
         8 . An isolated  Neisseria meningitidis  Hsp70 polypeptide.  
     
     
         9 . An isolated Hsp70 polypeptide comprising the amino acid sequence of according to FIG. 6 or variants thereof.  
     
     
         10 . An isolated Hsp70 polypeptide that is able to selectively bind to an antibody specific for a  Neisseria meningitidis  Hsp70.  
     
     
         11 . The isolated Hsp70 polypeptide according to  claim 9  wherein the isolated Hsp70 polypeptide is fused to an additional polypeptide to create a fusion protein.  
     
     
         12 . An isolated Hsp70 polypeptide comprising at least 8 contiguous amino acids selected from amino acid residues of FIG. 6, wherein the Hsp70 polypeptide is able to bind to a major histocompatibility complex.  
     
     
         13 . The isolated Hsp70 polypeptide according to  claim 12  wherein binding to the major histocompatibility complex elicits or enhances an immune response to  Neisseria meningitidis  in a human being.  
     
     
         14 . The isolated Hsp70 polypeptide according to  claim 12 , wherein the Hsp70 polypeptide is obtained by proteolytic cleavage.  
     
     
         15 . The isolated Hsp70 polypeptide according to  claim 12 , wherein the Hsp70 polypeptide is obtained by chemical synthesis.  
     
     
         16 . The isolated Hsp70 polypeptide according to  claim 12 , wherein the Hsp70 polypeptide is obtained by expression in a transformed host cell containing a nucleic acid molecule encoding the Hsp70 polypeptide or portion thereof.  
     
     
         17 . An isolated Hsp70 polypeptide comprising an amino acid sequence having at least 95% homology to the Hsp70 polypeptide of FIG. 6 and which is able to selectively bind to an antibody specific for a  Neisseria meningitidis  Hsp70, wherein percent homology is determined according to an algorithm incorporated in a protein database search program used in BLAST™ or DNA Star Megalign™.  
     
     
         18 . An isolated Hsp70 polypeptide wherein the polypeptide is an expression product of a transformed host cell containing an isolated nucleic acid molecule according to any one of claims  1 - 7 .  
     
     
         19 . A vector containing an isolated nucleic acid molecule according to any one of claims  1 - 7 .  
     
     
         20 . The vector according to  claim 19  wherein the vector is an expression vector.  
     
     
         21 . The vector according to  claim 20  further comprising a selectable or identifiable marker and wherein the promoter is a constitutive or an inducible promoter.  
     
     
         22 . A host cell containing a vector according to  claim 19 .  
     
     
         23 . The host cell of  claim 22  wherein the host cell is selected from the group consisting of a bacterial cell, a mammalian cell, a yeast cell, a plant cell and an insect cell.  
     
     
         24 . A composition comprising an Hsp70 polypeptide according to any one of claims  8 - 17  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         25 . A composition comprising a Hsp70polypeptide according to  claim 18  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         26 . The composition of  claim 24  wherein the pharmaceutically acceptable carrier or diluent is suitable for at least one of: systemic administration, oral administration, intranasal administration or parenteral administration.  
     
     
         27 . The composition of  claim 25  wherein the pharmaceutically acceptable carrier or diluent is suitable for at least one of: systemic administration, oral administration, intranasal administration or parenteral administration.  
     
     
         28 . A method for eliciting or enhancing an immune response in a mammal against  Neisseria meningitidis,  comprising administering to the mammal in an amount effective to elicit or enhance the response, an Hsp70 polypeptide according to any one of claims  8 - 17  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         29 . A method for eliciting or enhancing an immune response in a mammal against  Neisseria meningitidis,  comprising administering to the mammal in an amount effective to elicit or enhance the response, an Hsp70 polypeptide according to  claim 18  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         30 . A method for eliciting or enhancing an immune response in a mammal against a target antigen comprising administering to the mammal the target antigen joined to an Hsp70 polypeptide according to any one of claims  8 - 17  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         31 . A method for eliciting or enhancing an immune response in a mammal against a target antigen comprising administering to the mammal the target antigen joined to an Hsp70 polypeptide according to  claim 18  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         32 . A method for eliciting or enhancing an immune response in a mammal to a polypeptide comprising administering to the mammal a fusion protein containing sequences of the polypeptide fused to sequences of an  Neisseria meningitidis  Hsp70 polypeptide in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         33 . A probe or PCR primer for detecting DNA encoding a  Neisseria meningitidis  Hsp70 comprising at least about 15 contiguous bases from any one of SEQ. ID NOS: 1-4, or a compliment thereof.  
     
     
         34 . A method for diagnosing the presence of a  Neisseria meningitidis  in a subject sample comprising: 
 obtaining a DNA fraction from the subject sample; and    performing a PCR amplification of the DNA fraction using at least one PCR primer comprised of at least about 15 contiguous bases from any one of SEQ. ID NOS: 1-4, or a compliment thereof.    
     
     
         35 . An isolated nucleic acid molecule encoding a  Aspergillus fumigatus  Hsp60 polypeptide.  
     
     
         36 . An isolated nucleic acid molecule selected from the group consisting of: 
 (a) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 5;    (b) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 5 from nucleotides 300-2234;    (c) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 5 from nucleotides 300-410. nucleotides 514-655 and nucleotides 724-2234;    (d) an isolated nucleic acid molecule comprising-the sequence of SEQ ID NO: 6;    (e) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 7;    (f) an isolated nucleic acid molecule complementary to any one of the nucleotides of SEQ ID NOS: 5 to 7 set forth in (a) through (e), respectively.    
     
     
         37 . An isolated nucleic acid molecule which is a variant of, or is substantially similar to, the nucleotide molecule according to  claim 36 .  
     
     
         38 . An isolated nucleic acid molecule comprising a nucleotide sequence that is identical to a segment of contiguous nucleotide bases comprising at least 25% of any one of SEQ ID NOS: 5-7; or to a complement thereof.  
     
     
         39 . An isolated nucleic acid molecule encoding Hsp60 comprising a nucleic acid sequence that encodes a polypeptide comprising any one of the polypeptides according to FIGS. 14, 16 or  18 ; or a variant Hsp60 that is at least 95% homologous to a polypeptide according to any one of FIGS. 14, 16 or  18 , wherein percent homology is determined according to an algorithm incorporated in a protein database search program used in BLAST™ or DNA Star Megalign™.  
     
     
         40 . The isolated nucleic acid molecule according to  claim 39  wherein the encoded polypeptide is able to selectively bind an antibody specific for a  Aspergillus fumigatus  Hsp60.  
     
     
         41 . An isolated nucleic acid molecule encoding at least 8 contiguous amino acids of an  Aspergillus fumigatus  Hsp60 polypeptide selected from amino acid residues according to FIG. 14, wherein the encoded  Aspergillus fumigatus  Hsp60 polypeptide is able to bind a major histocompatibility complex.  
     
     
         42 . An isolated  Aspergillus fumigatus  Hsp60 polypeptide.  
     
     
         43 . An isolated Hsp60 polypeptide comprising the amino acid sequence according to FIG. 14, or variants thereof.  
     
     
         44 . An isolated Hsp60 polypeptide that is able to selectively bind to an antibody specific for a  Aspergillus fumigatus  Hsp60.  
     
     
         45 . The isolated Hsp60 polypeptide according to  claim 43  wherein the isolated Hsp60 polypeptide is fused to an additional polypeptide to create a fusion protein.  
     
     
         46 . An isolated Hsp70 polypeptide comprising at least 8 contiguous amino acids selected from amino acid residues selected from amino acid residues of FIG. 14, wherein the Hsp60 polypeptide is able to bind to a major histocompatibility complex.  
     
     
         47 . The isolated Hsp60 polypeptide according to  claim 46  wherein binding to the major histocompatibility complex elicits or enhances an immune response to  Aspergillus fumigatus  in a human being.  
     
     
         48 . The isolated Hsp60 polypeptide according to  claim 46 , wherein the Hsp60 polypeptide is obtained by proteolytic cleavage  
     
     
         49 . The isolated Hsp60 polypeptide according to  claim 46 , wherein the Hsp60 polypeptide is obtained by chemical synthesis.  
     
     
         50 . The isolated Hsp60 polypeptide according to  claim 46 , wherein the Hsp60 polypeptide is obtained by expression in a transformed host cell containing a nucleic acid molecule encoding the Hsp60 polypeptide or portion thereof.  
     
     
         51 . An isolated Hsp60 polypeptide comprising an amino acid sequence having at least 95% homology to the Hsp60 polypeptide of FIG. 14, and which is able to selectively bind to an antibody specific for an  Aspergillus fumigatus  Hsp60, wherein percent homology is determined according to an algorithm incorporated in a protein database search program used in BLAST™ or DNA Star Megalign™.  
     
     
         52 . An isolated Hsp60 polypeptide wherein the polypeptide is an expression product of a transformed host cell containing an isolated nucleic acid molecule according to any one of claims  35 - 41 .  
     
     
         53 . A vector containing an isolated nucleic acid molecule according to any one of claims  35 - 41 .  
     
     
         54 . The vector according to  claim 53  wherein the vector is an expression vector.  
     
     
         55 . The vector according to  claim 54  further comprising a selectable or identifiable marker and wherein the promoter is a constitutive or an inducible promoter.  
     
     
         56 . A host cell containing a vector according to  claim 53 .  
     
     
         57 . The host cell according to  claim 56  wherein the host cell is selected from the group consisting of a bacterial cell, a mammalian cell, a yeast cell, a plant cell and an insect cell.  
     
     
         58 . A composition comprising an Hsp60 polypeptide according to any one of claims  35 - 41  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         59 . A composition comprising a Hsp60 polypeptide according to  claim 47  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         60 . The composition of  claim 58  wherein the pharmaceutically acceptable carrier or diluent is suitable for at least one of: systemic administration, oral administration, intranasal administration or parenteral administration.  
     
     
         61 . The composition of  claim 59  wherein the pharmaceutically acceptable carrier or diluent is suitable for at least one of: systemic administration, oral administration, intranasal administration or parenteral administration.  
     
     
         62 . A method for eliciting or enhancing an immune response in a mammal against  Aspergillus fumigatus,  comprising administering to the mammal in an amount effective to elicit or enhance the response, an Hsp60 polypeptide according to any one of claims  35 - 41  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         63 . A method for eliciting or enhancing an immune response in a mammal against  Aspergillus fumigatus,  comprising administering to the mammal in an amount effective to elicit or enhance the response, an Hsp60 polypeptide according to  claim 47  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         64 . A method for eliciting or enhancing an immune response in a mammal against a target antigen comprising administering to the mammal the target antigen joined to an Hsp60 polypeptide according to any one of claims  35 - 41  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         65 . A method for eliciting or enhancing an immune response in a mammal against a target antigen comprising administering to the mammal the target antigen joined to an Hsp60 polypeptide according to  claim 47  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         66 . A method for eliciting or enhancing an immune response in a mammal to a polypeptide comprising administering to the mammal a fusion protein containing sequences of the polypeptide fused to sequences of an  Aspergillus fumigatus  Hsp60 polypeptide in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         67 . A probe or PCR primer for detecting DNA encoding a  Aspergillus fumigatus  Hsp60 comprising at least about 15 contiguous bases from any one of SEQ. ID NOS: 5-7, or a compliment thereof.  
     
     
         68 . A method for diagnosing the presence of a  Aspergillus fumigatus  in a subject sample comprising: 
 obtaining a DNA fraction from the subject sample; and    performing a PCR amplification of the DNA fraction using at least one PCR primer comprised of at least about 15 contiguous bases from any one of SEQ. ID NOS: 5-7, or a compliment thereof.    
     
     
         69 . An isolated nucleic acid molecule encoding a  Candida glabrata  Hsp60.  
     
     
         70 . An isolated nucleotide molecule selected from the group consisting of: 
 (a) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 8;    (b) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 8 from nucleotides 258-1964;    (c) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 9;    (d) an isolated nucleic acid molecule comprising the sequence of SEQ ID NO: 10;    (e) an isolated nucleic acid molecule complementary to any one of the nucleotides of SEQ ID NOS: 8 to 10 set forth in (a) through (d), respectively.    
     
     
         71 . An isolated nucleic acid molecule which is a variant of, or is substantially similar to, the nucleotide molecule according to  claim 70 .  
     
     
         72 . An isolated nucleic acid comprising a nucleotide sequence that is identical to a segment of contiguous nucleotide bases comprising at least 25% of any one of SEQ ID NOS: 8-10, or to a complement thereof.  
     
     
         73 . An isolated nucleic acid molecule encoding Hsp60 comprising a nucleic acid sequence that encodes a polypeptide comprising any one of the polypeptides according to FIGS. 21, 23, or  25 , or a variant Hsp60 that is at least 95% homologous to a polypeptide according to any one of FIGS. 21, 23, or  25 , wherein percent homology is determined according to an algorithm incorporated in a protein database search program used in BLAST™ or DNA Star Megalign™.  
     
     
         74 . The isolated nucleic acid molecule according to  claim 73  wherein the encoded polypeptide is able to selectively bind an antibody specific for a  Candida glabrata  Hsp60.  
     
     
         75 . An isolated nucleic acid molecule encoding at least 8 contiguous amino acids of a  Candida glabrata  Hsp60 polypeptide selected from amino acid residues according to FIG. 21, wherein the encoded  Candida glabrata  Hsp60 polypeptide is able to bind to a major histocompatibility complex.  
     
     
         76 . An isolated  Candida glabrata  Hsp60 polypeptide.  
     
     
         77 . An isolated Hsp60 polypeptide comprising the amino acid sequence of FIG. 21, or variants thereof.  
     
     
         78 . An isolated Hsp60 polypeptide the that is able to selectively bind to an antibody specific for a  Candida glabrata  Hsp60.  
     
     
         79 . The isolated Hsp60 polypeptide according to  claim 77  wherein the isolated Hsp70 polypeptide is fused to an additional polypeptide to create a fusion protein.  
     
     
         80 . An isolated Hsp60 polypeptide comprising at least 8 contiguous amino amino acids selected from amino acid residues according to FIG. 21, wherein the Hsp60 polypeptide is is able to bind to a major histocompatibility complex.  
     
     
         81 . The isolated Hsp60 polypeptide according to  claim 80  wherein binding to the major histocompatibility complex elicits or enhances an immune response to  Candida glabrata  in a human being.  
     
     
         82 . The isolated Hsp60 polypeptide according to  claim 80  wherein the Hsp60 polypeptide is obtained by proteolytic cleavage.  
     
     
         83 . The isolated Hsp60 polypeptide according to  claim 80 , wherein the Hsp70 polypeptide is obtained by chemical synthesis.  
     
     
         84 . The isolated Hsp60 polypeptide according to  claim 80 , wherein the Hsp70 polypeptide is obtained by expression in a transformed host cell containing a nucleic acid molecule encoding the Hsp60 polypeptide or portion thereof.  
     
     
         85 . An isolated Hsp60 polypeptide comprising an amino acid sequence having at least 95% homology to the Hsp60 polypeptide of FIG. 21, and which is able to selectively bind to an antibody specific for a  Candida glabrata  Hsp60, wherein percent homology is determined according to an algorithm incorporated in a protein database search program used in BLAST™ or DNA Star Megalign™.  
     
     
         86 . An isolated Hsp60 polypeptide wherein the polypeptide is an expression product of a transformed host cell containing an isolated nucleic acid molecule according to any one of claims  69 - 75 .  
     
     
         87 . A vector containing an isolated nucleic acid molecule according to any one of claims  69 - 75 .  
     
     
         88 . The vector according to  claim 87  wherein the vector is an expression vector.  
     
     
         89 . The vector according to  claim 88  further comprising a selectable or identifiable marker and wherein the promoter is a constitutive or an inducible promoter.  
     
     
         90 . A host cell containing a vector according to  claim 87 .  
     
     
         91 . The host cell of  claim 90  wherein the host cell is selected from the group consisting of a bacterial cell, a mammalian cell, a yeast cell, a plant cell and an insect cell.  
     
     
         92 . A composition comprising an Hsp60 polypeptide according to any one of claims  76 - 85  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         93 . A composition comprising a Hsp60 polypeptide according to  claim 86  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         94 . The composition of  claim 92  wherein the pharmaceutically acceptable carrier or diluent is suitable for at least one of: systemic administration, oral administration, intranasal administration or parenteral administration.  
     
     
         95 . The composition of  claim 92  wherein the pharmaceutically acceptable carrier or diluent is suitable for at least one of: systemic administration, oral administration, intranasal administration or parenteral administration.  
     
     
         96 . A method for eliciting or enhancing an immune response in a mammal against  Candida glabrata,  comprising administering to the mammal an in an amount effective to elicit or enhance the response, an Hsp60 polypeptide according to any one of claims  76 - 85  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         97 . A method for eliciting or enhancing an immune response in a mammal against  Candida glabrata,  comprising administering to the mammal an in an amount effective to elicit or enhance the response, an Hsp60 polypeptide according to  claim 86  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         98 . A method for eliciting or enhancing an immune response in a mammal against a target antigen comprising administering to the mammal the target antigen joined to an Hsp60 polypeptide according to any one of claims  76 - 85  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         99 . A method for eliciting or enhancing an immune response in a mammal against a target antigen comprising administering to the mammal the target antigen joined to an Hsp70 polypeptide according to  claim 86  in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         100 . A method for eliciting or enhancing an immune response in a mammal to a polypeptide comprising administering to the mammal a fusion protein containing sequences of the polypeptide fused to sequences of an  Candida glabrata  Hsp60 polypeptide in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         101 . A probe or PCR primer for detecting DNA encoding a  Candida glabrata  Hsp60 comprising at least about 15contiguous bases from any one of SEQ. ID NOS: 8-10, or a compliment thereof.  
     
     
         102 . A method for diagnosing the presence of  Candida glabrata  in a subject sample comprising: 
 obtaining a DNA fraction from the subject sample; and    performing a PCR amplification of the DNA fraction using at least one PCR primer comprised of at least about 15 contiguous bases from any one of SEQ. ID NOS: 8-10, or a compliment thereof.