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Isolation and characterization of a novel major Dermatophagoides pteronyssinus mite allergen, Der p 11
J ALLERGY CLIN IMMUNOL VOLUME 113, NUMBER 2
Abstracts S225
798
Isolation and Characterization of a Novel Major Dermatophagoides pteronyssinus Mite Allergen, Der p 11
L. C. Tsai1, P. L. Chao1, R. B. Tang2, M. W. Hung1, H. D. Shen1, S. H. Han1; of Medical and Education, Paipei-Veterans General Hospital, Taipei, TAIWAN REPUBLIC OF CHINA, 2Deprtment of Pediatrics, PaipeiVeterans General Hospital, Taipei, TAIWAN REPUBLIC OF CHINA. RATIONALE: Dermatophagoides pteronyssinus (Dp) and D. farinae (Df) mites have been regarded as a major source of the indoor allergens. To date, two groups of high molecular weight (mw) dust-mite allergens to human have been identified. Isolation of high mw Dp mite allergens is 1Department
MONDAY
important in characterizing the complete spectrum of mite IgE specificities and providing a suitable substance for clinical application. METHODS: A full-length Der p 11 gene was isolated by Dp cDNA library immunoscreening, 5’-3’ rapid amplification of cDNA ends (RACE) and polymerase chain reaction (PCR). Sequence analysis and alignment were scanned with a combination of CGC and BLAST program package. RESULTS: The Der p 11 gene is a 2965-bp cDNA gene with 2625-bp open reading frame coding for an 875-amino acid protein. The protein sequence of the newly recognized Dp allergen exhibited significant homology with other invertebrate paramyosins. The whole cDNA insert and PCR-derived fragments were generated and expressed in E. coli. The recombinant proteins including Der p 11 and its derivative peptides (p1-p4) were used for IgE binding immunoassay. An in vitro IgE-binding study showed that 75% (15/20), 65% (13/20), 76% (38/50), 60% (18/30), 73.3% (22/30), 66.7% (20/30), 63.3% (19/30) and 6.7% (2/30) in mite-sensitive asthmatic sera reacted positively with rDer p 2, rDer f 11, rDer p 11, p1, p2, p3, p4 and GST, respectively. IgE reactivity to rDer p11 fragments frequently reacted even in those patients with MAST(Dp)-negative sera. CONCLUSIONS: Der p 11 is the homology of Der f 11 exhibiting high IgE reactivity to allergic sera from asthmatic children, potentially important house dust mite Dp allergen. Funding: Taipei-Veterans General Hospital
Abstracts S225
798
Isolation and Characterization of a Novel Major Dermatophagoides pteronyssinus Mite Allergen, Der p 11
L. C. Tsai1, P. L. Chao1, R. B. Tang2, M. W. Hung1, H. D. Shen1, S. H. Han1; of Medical and Education, Paipei-Veterans General Hospital, Taipei, TAIWAN REPUBLIC OF CHINA, 2Deprtment of Pediatrics, PaipeiVeterans General Hospital, Taipei, TAIWAN REPUBLIC OF CHINA. RATIONALE: Dermatophagoides pteronyssinus (Dp) and D. farinae (Df) mites have been regarded as a major source of the indoor allergens. To date, two groups of high molecular weight (mw) dust-mite allergens to human have been identified. Isolation of high mw Dp mite allergens is 1Department
MONDAY
important in characterizing the complete spectrum of mite IgE specificities and providing a suitable substance for clinical application. METHODS: A full-length Der p 11 gene was isolated by Dp cDNA library immunoscreening, 5’-3’ rapid amplification of cDNA ends (RACE) and polymerase chain reaction (PCR). Sequence analysis and alignment were scanned with a combination of CGC and BLAST program package. RESULTS: The Der p 11 gene is a 2965-bp cDNA gene with 2625-bp open reading frame coding for an 875-amino acid protein. The protein sequence of the newly recognized Dp allergen exhibited significant homology with other invertebrate paramyosins. The whole cDNA insert and PCR-derived fragments were generated and expressed in E. coli. The recombinant proteins including Der p 11 and its derivative peptides (p1-p4) were used for IgE binding immunoassay. An in vitro IgE-binding study showed that 75% (15/20), 65% (13/20), 76% (38/50), 60% (18/30), 73.3% (22/30), 66.7% (20/30), 63.3% (19/30) and 6.7% (2/30) in mite-sensitive asthmatic sera reacted positively with rDer p 2, rDer f 11, rDer p 11, p1, p2, p3, p4 and GST, respectively. IgE reactivity to rDer p11 fragments frequently reacted even in those patients with MAST(Dp)-negative sera. CONCLUSIONS: Der p 11 is the homology of Der f 11 exhibiting high IgE reactivity to allergic sera from asthmatic children, potentially important house dust mite Dp allergen. Funding: Taipei-Veterans General Hospital