Blomia tropicalis Blo t 5 and Blo t 21 recombinant allergens might confer higher specificity to serodiagnostic assays than whole mite extract

BMC Immunology

Neuza Maria Alcantara-Neves⁴  Lain Carlos Pontes-de-Carvalho¹  Carina Silva Pinheiro⁴  Álvaro Augusto Cruz²  Givaneide dos Santos Lima⁵  Mariese Conceição Alves dos Santos⁶  Filipe Adriano Borba Felipe⁴  João Carlos Marques Ponte⁴  Franklin Barbalho Magalhães⁷  Osvaldo Pompílio de Melo-Neto³  Kellyanne dos Anjos Carvalho⁴ 

[1] Centro de Pesquisas Gonçalo Moniz, Fundação Oswaldo Cruz, Salvador, Bahia, Brazil;ProAR – Nucleo de Excelência em Asma, Universidade Federal da Bahia, Salvador, Bahia, Brazil;Centro de Pesquisas Aggeu Magalhães, Fundação Oswaldo Cruz, Recife, Pernambuco, Brazil;Laboratório de Alergia e Acarologia, Instituto de Ciências da Saúde, Universidade Federal da Bahia, Avenida Reitor Miguel Calmon, sem nº, Canela, Salvador, Bahia, CEP 40110-100, Brazil;Centro de Estudos de Alergias Respiratórias, Salvador, Bahia, Brazil;Instituto de Matemática, Universidade Federal da Bahia, Salvador, Bahia, Brazil;Associação Caruaruense de Ensino Superior, Caruaru, Pernambuco, Brazil

关键词: Specificity;    Sensitivity;    Ascaris lumbricoides;    Cross-reactivity;    Immunodiagnosis;    Recombinant allergens;    Blomia tropicalis;   

Others  :  1077834
DOI  :  10.1186/1471-2172-14-11

 received in 2012-09-19, accepted in 2013-02-21,  发布年份 2013

【 摘 要 】

Background

Blomia tropicalis is a dust mite and an important source of allergens in tropical regions. Up to now, the assays to diagnose atopy to this mite use whole body extract as antigens. However, anti-B. tropicalis IgE antibodies cross-react with Ascaris lumbricoides antigens, hindering the diagnosis of allergy to this mite. In this study, B. tropicalis recombinant allergens were evaluated with the purpose of developing an immunodiagnostic assay for allergy to this mite with greater specificity than those commercially available.

Methods

Two B. tropicalis allergens (Blo t 5 and Blo t 21) were cloned into a plasmidial expression vector, expressed in Escherichia coli and purified by affinity chromatography. Sixty-three sera containing anti-B. tropicalis extract (BtE) IgE antibodies were used to investigate IgE reactivity to the recombinant Blot 5 and 21 allergens. Inhibition assays with 20 sera pre-adsorbed with A. lumbricoides extract were performed using rBlo t 5, rBlo t 21, and BtE as antigens. All the assays were carried using indirect ELISA.

Results

Eighty-two point nine percent and 80.0% of the sera with anti-BtE antibodies from 35 children reacted with rBlo t 5 and rBlo t 21, respectively, whereas 92.8% and 89.3% of the 28 sera with anti-BtE antibodies from adult asthma patients reacted with the same allergens, and 96.4% of these sera reacted with a mixture of rBlo t 5 and rBlo t 21. In an inhibition ELISA, the absorption of sera by A. lumbricoides extract affected less the reaction with rBlo t 5 and rBlo t 21 than with BtE.

Conclusions

The rBlo t 5 and rBlo t 21 allergens contain important epitopes recognized by IgE antibodies of individuals allergic to B. tropicalis antigens. Moreover, the assays using the recombinant allergens had lower IgE cross-reactivity with A. lumbricoides antigens, a fact which would confers higher specificity to serodiagnostic assays than the crude mite extract. However, additional recombinant allergens should be evaluated in order to reach the same sensitivity of the commercially available assays based on mite extract.

【 授权许可】

   
2013 Carvalho et al.; licensee BioMed Central Ltd.

【 图 表 】

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