Original Articles: Mechanisms Of AllergyAllergens of wild house dust mites: Environmental Der p 1 and Der p 2 sequence polymorphisms☆,☆☆
Section snippets
Materials
D pteronyssinus mites were collected in Perth, Western Australia, and by Dr Euan Tovey from Sydney. They were collected by vacuuming carpet or soft furnishings for 2 to 3 minutes with filter paper in a filter apparatus attached to the head of a vacuum cleaner pipe. The dust collected in the filter from the Perth houses was placed in a mesh container secured at the top of a funnel; a 1.5-mL Eppendorf was placed at the base of the funnel and used to collect the mites after exposure of the dust to
Wild mite reverse transcriptase PCR
The first batch of mites to be amplified were the dried mite cultures from Sydney. For the remaining mites, a reverse transcriptase PCR method was used. RNA was successfully extracted from every sample of mites, even when only a single mite was used as starting material. The RNA extracted from a single mite was sufficient for the synthesis of good yields of first-strand cDNA with gene-specific primers.
Der p 2 wild mite cDNA sequences
Nine Der p 2 cDNAs were isolated from the Perth wild mites. The sequences revealed 16
Discussion
Der p 1 and Der p 2 are major allergic specificities and targets for new immunotherapeutic strategies such as the injection of peptides, modified allergens, or DNA vaccines. cDNA has provided the information to pursue these objectives but has also revealed that they exhibit a high frequency of allelic polymorphism. This study examined which variants found in the environment could best represent the spectrum of polymorphic proteins that induce sensitization. It was found that although the
Acknowledgements
We would like to thank Dr Euan Tovey, from the Institute of Respiratory Medicine, University of Sydney, Australia, for collecting and preparing the dried house dust mite samples from the Sydney homes.
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Hierarchy and molecular properties of house dust mite allergens
2015, Allergology InternationalCitation Excerpt :Der f 2 can bind lipopolysaccharide (LPS) at high affinity with structural changes similar to those occurring for its MD-2 homologue.28 Allelic variants of Der p 220,21,24,29 and Der f 221,24,30 show a pattern indicating different genetic lineages that might be important for allergy. For D. pteronyssinus the amino acids 40, 47, 111 and 114 VTMD (in single letter amino acid notation) in Der p 2.0101 can be substituted with varying penetrance to LSLN respectively with Der p 2.0104 having all four substitutions.
House dust mites possess a polymorphic, single domain putative peptidoglycan d,l endopeptidase belonging to the NlpC/P60 Superfamily
2015, FEBS Open BioCitation Excerpt :In addition to identifying polymorphisms in D. pteronyssinus, a conservative polymorphism in the D. farinae homologue was also identified using the reported unplaced genomic scaffold (KN266412) [19]. These findings are consistent with those seen with other mite proteins, in particular the cysteine protease, trypsin and the MD-related group 2 protein [28,29]. This study also showed that there were no introns in the D. pteronyssinus lytFM gene which contrasts with that seen in D. farinae [19], where a single intron was observed in the leader sequence of the gene rather than in the region encoding the mature protein.
Recombinant house dust mite allergens
2014, MethodsCitation Excerpt :HDM allergens are simpler than most pollen allergens because the dominant allergens are encoded by single genes and not gene families. The sequences are however very polymorphic [59] with approximately half of 48 sequences of Der p 1 analysed by Piboonpocanun et al. being different [60]. The sequences differed from each by 1–5 residues but mostly with sporadic substitutions so that the predominant core sequence can easily represented by a single sequence.
House dust mite allergens in asthma and allergy
2010, Trends in Molecular MedicineThe rules of variation: Amino acid exchange according to the rotating circular genetic code
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2009, Molecular Immunology
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Supported by the Australian National Health and Medical Research Council.
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Reprint requests: Wayne R. Thomas, PhD, TVW Telethon Institute for Child Health Research, PO Box 855, West Perth, Western Australia 6872, Australia.