Cloning and sequencing of allergens of dermatophagoides (house dust mite)

Abstract

Isolated DNA encoding allergens of Dermatophagoides (house dust mites) particularly of the species Dermatophagoides farinae and Dermatophagolides pteronyssinus, which are protein allergens or peptides which include at least one epitope of the protein allergen. In particular, DNA encoding two major D. farinae allergens, Der f I and Der f II and DNA encoding a D. pteronyssinus allergen, Der p I. In addition, the proteins or peptides encoded by the isolated DNA, their use as diagnostic and therapeutic reagents and methods of diagnosing and treating sensitivity to house dust mite allergens.

1 . (canceled)  
     
     
         2 . A method of producing an isolated protein of  Dermatophagoides pteronyssinus  comprising the steps of: 
 a) culturing a host cell transformed with a nucleic acid encoding a protein allergen of  Dermatophagoides pteronyssinus, Der p  I, comprising the amino acid sequence shown in  FIG. 7 , in an appropriate medium to produce a mixture of cells and medium containing said protein allergen; and    b) purifying said mixture to produce isolated  Der p  I protein allergen.    
     
     
         3 . The method of  claim 2  wherein the nucleic acid comprises a nucleotide sequence selected from the group consisting of the nucleotide sequence shown in  FIG. 7 , the coding region of the nucleotide sequence shown in  FIG. 7 , and a nucleotide sequence which hybridizes under high or low stringency conditions to a complementary strand of the nucleotide sequence shown in  FIG. 7 .  
     
     
         4 . The method of  claim 3 , wherein the high stringency conditions comprise hybridization at Tm-20 followed by at least one post-hybridization wash at Tm-12.  
     
     
         5 . The method of  claim 2 , wherein the nucleic acid comprises the cDNA insert of phage clone λgt11 p1(13T), ATCC Deposit No. 69338.  
     
     
         6 . A method of producing an isolated protein of  Dermatophagoides pteronyssinus  comprising the steps of: 
 a) culturing a host cell transformed with a nucleic acid encoding a protein allergen of  Dermatophagoides pteronyssinus, Der p  I, and selected from the group consisting of the nucleotide sequence shown in  FIG. 7 , the coding region of the nucleotide sequence shown in  FIG. 7 , and a nucleotide sequence which hybridizes under high or low stringency conditions to a complementary strand of the nucleotide sequence shown in  FIG. 7 , in an appropriate medium to produce a mixture of cells and medium containing said protein allergen; and    b) purifying said mixture to produce isolated  Der p  I protein allergen.    
     
     
         7 . A method of producing an isolated antigenic peptide of  Dermatophagoides pteronyssinus  comprising the steps of: 
 a) culturing a host cell transformed with a nucleic acid encoding an antigenic peptide of  Dermatophagoides pteronyssinus, Der p  I, comprising a portion of the amino acid sequence shown in  FIG. 7 , in an appropriate medium to produce a mixture of cells and medium containing said peptide; and    b) purifying said mixture to produce isolated  Der p  I peptide.    
     
     
         8 . The method of  claim 7  wherein the nucleic acid comprises a portion of a nucleotide sequence selected from the group consisting of the nucleotide sequence shown in  FIG. 7 , the coding region of the nucleotide sequence shown in  FIG. 7 , and a nucleotide sequence which hybridizes under high or low stringency conditions to a complementary strand of the nucleotide sequence shown in  FIG. 7 .  
     
     
         9 . The method of  claim 7 , wherein the high stringency conditions comprise hybridization at Tm-20 followed by at least one post-hybridization wash at Tm-12.  
     
     
         10 . The method of  claim 7 , wherein the nucleic acid comprises a portion of the cDNA insert of phage clone λgt11 p1(13T), ATCC Deposit No. 69338.  
     
     
         11 . A method of producing an isolated protein of  Dermatophagoides pteronyssinus  comprising the steps of: 
 a) culturing a host cell transformed with a nucleic acid encoding an antigenic peptide of  Dermatophagoides pteronyssinus, Der p  I, and selected from the group consisting of a portion of the nucleotide sequence shown in  FIG. 7 , a portion of the coding region of the nucleotide sequence shown in  FIG. 7 , and a portion of the a nucleotide sequence which hybridizes under high or low stringency conditions to a complementary strand of the nucleotide sequence shown in  FIG. 7 , in an appropriate medium to produce a mixture of cells and medium containing said protein allergen; and    b) purifying said mixture to produce isolated  Der p  I protein allergen.