Identification of &lt;i&gt;Blomia tropicalis&lt;/i&gt; Allergen Blo t 5 by cDNA Cloning

Arruda, Lia Ferraz de; Fernández-Caldas, Enrique; Ck, Naspitz; Montealegre, Federico; Ld, Vailes; Chapman, Martin D.

doi:10.1159/000237080

Cited by 42 publications

(50 citation statements)

References 5 publications

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“…Tsai (Veterans General Hospital, Taipei, Taiwan). rBlo t 5-GST was purified from E. coli as previously described [8]. …”

Section: Methodsmentioning

confidence: 99%

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cDNA Cloning and Expression of Blo t 11, the <i>Blomia tropicalis</i> Allergen Homologous to Paramyosin

Ramos¹,

Cheong

Lee³

et al. 2001

Int Arch Allergy Immunol

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Blomia tropicalis is an important mite species in many parts of the world and the most predominant mite species in tropical countries. The prevalence of sensitization to this species has probably been underestimated because commercial extracts are largely unavailable. Identification and characterization of B. tropicalis allergens is an important step toward understanding the role of this species in allergic sensitization and could provide appropriate reagents for diagnostic and therapeutic procedures. This paper describes the isolation, sequence analysis, expression and allergenicity of a cDNA gene coding for a B. tropicalis allergen with homology to paramyosin, a high-molecular-weight allergen previously identified in Dermatophagoides farinae. The full-length Blo t 11 cDNA gene was isolated by cDNA library screening, 5′-rapid amplification of cDNA ends and long-distance PCR. Sequence analysis was performed with a combination of CLUSTAL W, CGC and BLAST program packages. The cDNA gene was expressed as a GST fusion protein in Escherichia coli and purified by affinity chromatography using the glutathione Sepharose column. Allergenicity of the rBlo t 11 was tested by human IgE dot blot immunoassay. Blo t 11 is a 3,111-bp cDNA gene with a 2,625-bp open reading frame coding for an 875-amino acid protein, exhibiting significant homology with different invertebrate paramyosins. The human IgE dot blot immunoassay showed that the rBlo t 11 reacted positively to 52% (33/63) of sera from asthmatic patients. Blo t 11 is the homolog of Der f 11 exhibiting potentially important allergenic activity.

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“…Tsai (Veterans General Hospital, Taipei, Taiwan). rBlo t 5-GST was purified from E. coli as previously described [8]. …”

Section: Methodsmentioning

confidence: 99%

“…However, despite the fact that more than 10 allergens had been cloned and characterized in Dermatophagoides spp. [6, 7], only 3 Bt allergens have been cloned, and their allergenicity established [8, 9, 10, 11]. These Bt allergens include Blo t 5 [8, 9], Blo t 13 (previously known as Bt 6) [10]and Blo t 12 (previously known as Bt 11a) [11].…”

Section: Introductionmentioning

confidence: 99%

cDNA Cloning and Expression of Blo t 11, the <i>Blomia tropicalis</i> Allergen Homologous to Paramyosin

Ramos¹,

Cheong

Lee³

et al. 2001

Int Arch Allergy Immunol

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“…Arruda et al (19) failed to observe a correlation between Blo t 5 and Der p 5 in eliciting IgE responses in Blomia tropicalis-exposed and -unexposed individuals, despite 43% identity at the amino acid level. Similarly, in a study on Malaysian and Taiwanese patients, Kuo et al (20) reported a low correlation of IgE reactivity between Blo t 5 and Der p 5 (Taiwanese, r=0.452; Malaysian, r=0.346) as well as limited cross-reactivity.…”

Section: Introductionmentioning

confidence: 95%

“…In tropical regions, however, the Blomia tropicalis allergen Blo t 5 causes reactions in 70-92% of patients with mite allergies, making it the predominant reaction-associated group-5 allergen in that region (19,20). Group-21 allergens, which are homologous with the group-5 allergens, have only been described in Blomia tropicalis and Dermatophagoides pteronyssinus (21,22).…”

Section: Introductionmentioning

confidence: 99%

Cross-reactivity between group-5 and -21 mite allergens from Dermatophagoides farinae, Tyrophagus putrescentiae and Blomia tropicalis

Kim

Jeong

et al. 2015

Molecular Medicine Reports

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Abstract. Group-5 and group-21 allergens, produced by house dust mites and storage mites are 36.6-55.8% identical in their sequences and are recognized by at least 50% of immunoglobulin (Ig)E from the sera of individuals allergic to dust mites. In the present study, recombinant group-5 and -21 allergens from three mite species, Dermatophagoides farinae (rDer f 5 and 21), Tyrophagus putrescentiae (rTyr p 5 and 21), and Blomia tropicalis (rBlo t 5 and 21), were purified from Escherichia coli, and the IgE reactivities and cross-reactivities of these allergen variants were assessed. The IgE binding frequencies of rDer f 5, rDer f 21, rTyr p 5, rTyr p 21, rBlo t and rBlo t 21 proteins were 64. 95, 65.98, 30.41, 41.24, 30.93 and 21.65%, respectively. The IgE reactivity of rDer f 5 correlated highly with that of rDer f 21 (r=0.733). rTyr p 5 exhibited the highest level of correlation with rTyr p 21 (r=0.950), while the correlation of rBlo t 5 with rBlo t 21 was the lowest observed (r=0.104). The binding of IgE to rDer f 5 and rDer f 21 was not inhibited by any allergens but themselves. While rDer f 5 inhibited only 60.3% of IgE binding to rBlo t 5, rDer f 21 exhibited a high inhibitory effect against rTyr p 5 (93.01%), rTyr p 21 (92.12%), rBlo t 5 (86.97%) and rBlo t 21 (70.30%), implying cross-reactivity among mite species. The results of the present study demonstrated that the majority of the IgE reactivity to group-5 and -21 storage mite allergens is due to cross-reaction. It is therefore imperative to develop an accurate, component-resolved diagnosis for dust mite allergies.

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“…In D. pteronyssinus, Der p 1 and 2 bound 50 -65% of the IgE Abs, and only around 10% of the IgE bound to each of Der p 4, 5, and 7 (8). In contrast, among the seven allergens evaluated in B. tropicalis, Blo t 2a, 3,5,10,11,12, and 13, Blo t 5 is by far the most important allergenic component and Blo t 5-specific IgE is found in 40 -60% of B. tropicalis-sensitive individuals (9). Blo t 5 contains a 17-residues signal peptide and a 117-residues mature protein, while Der p 5 is made up of 19-residues signal peptide and a 113-residues mature protein (10).…”

mentioning

confidence: 91%

Nuclear Magnetic Resonance Structure and IgE Epitopes of Blo t 5, a Major Dust Mite Allergen

Chan¹,

Ong²,

Gao³

et al. 2008

The Journal of Immunology

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A high incidence of sensitization to Blomia tropicalis, the predominant house dust mite species in tropical regions, is strongly associated with allergic diseases in Singapore, Malaysia, and Brazil. IgE binding to the group 5 allergen, Blo t 5, is found to be the most prevalent among all B. tropicalis allergens. The NMR structure of Blo t 5 determined represents a novel helical bundle structure consisting of three antiparallel α-helices. Based on the structure and sequence alignment with other known group 5 dust mite allergens, surface-exposed charged residues have been identified for site-directed mutagenesis and IgE binding assays. Four charged residues, Glu76, Asp81, Glu86, and Glu91 at around the turn region connecting helices α2 and α3 have been identified to be involved in the IgE binding. Using overlapping peptides, we have confirmed that these charged residues are located on a major putative linear IgE epitope of Blo t 5 from residues 76–91 comprising the sequence ELKRTDLNILERFNYE. Triple and quadruple mutants have been generated and found to exhibit significantly lower IgE binding and reduced responses in skin prick tests. The mutants induced similar PBMC proliferation as the wild-type protein but with reduced Th2:Th1 cytokines ratio. Mass screening on a quadruple mutant showed a 40% reduction in IgE binding in 35 of 42 sera of atopic individuals. Findings in this study further stressed the importance of surface-charged residues on IgE binding and have implications in the cross-reactivity and use of Blo t 5 mutants as a hypoallergen for immunotherapy.

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Identification of <i>Blomia tropicalis</i> Allergen Blo t 5 by cDNA Cloning

Cited by 42 publications

References 5 publications

cDNA Cloning and Expression of Blo t 11, the <i>Blomia tropicalis</i> Allergen Homologous to Paramyosin

cDNA Cloning and Expression of Blo t 11, the <i>Blomia tropicalis</i> Allergen Homologous to Paramyosin

Cross-reactivity between group-5 and -21 mite allergens from Dermatophagoides farinae, Tyrophagus putrescentiae and Blomia tropicalis

Nuclear Magnetic Resonance Structure and IgE Epitopes of Blo t 5, a Major Dust Mite Allergen

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