Molecular Cloning and Characterization of Tropomyosin, a Major Allergen of Chironomus kiiensis, a Dominant Species of Nonbiting Midges in Korea

Kyoung Yong Jeong, Hye Yung Yum, In Yong Lee, Han Il Ree, Chein Soo Hong, Dong Soo Kim, Tai Soon Yong

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Chironomids are widely and abundantly distributed in the vicinity of standing waters. Larvae of Chironomus and some other genera are known to contain hemoglobins, which have been described as a major allergen, and the adults that have no hemoglobins also have been reported to contain allergens. In this study, we tried to establish the role of chironomid allergy and characterize the allergen of Chironomus kiiensis adults. Skin tests using C. kiiensis adult extracts were performed on patients with allergic symptoms. A cDNA library of C. kiiensis adults was screened with C. kiiensis immune mouse sera to identify allergens, and results were confirmed using skin test-positive human sera. Recombinant allergen was expressed in Escherichia coli and purified by affinity chromatography using nickel-nitrilotriacetic acid agarose to investigate its allergenic properties. Out of 275 allergic patients 14.2% showed a positive reaction to C. kiiensis adult crude extracts in the skin test. The tropomyosin was cloned by immunoscreening and expressed in Escherichia coli. C. kiiensis tropomyosin has a high homology at the amino acid level with tropomyosins which were previously known to be allergens in various arthropods (Periplaneta americana, 86.3%; Panulirus stimpson, 78.9%; Dermatophagoides pteronyssinus, 76.5%). Specific immunoglobulin E antibodies reacting to recombinant tropomyosin were detected in 17 (81%) of 21 patients whose skin test results were positive. Cross-reactivity against house dust mites and other insects was noticed with mouse anti-recombinant tropomyosin immune serum. C. kiiensis adults were shown to be an important source of inhalant allergens in Korea. Molecular cloning of C. kiiensis tropomyosin was performed and IgE reactivity was demonstrated using skin test-positive human sera. Recombinant tropomyosin will be useful for further studies or clinical applications.

Original languageEnglish
Pages (from-to)320-324
Number of pages5
JournalClinical and Diagnostic Laboratory Immunology
Volume11
Issue number2
DOIs
Publication statusPublished - 2004 Mar 1

Fingerprint

Chironomidae
Tropomyosin
Cloning
Molecular Cloning
Korea
Allergens
Skin Tests
Skin
Escherichia coli
Immunoglobulin E
Immune Sera
Hemoglobins
Palinuridae
Dermatophagoides pteronyssinus
Periplaneta
Allergies
Affinity chromatography
Pyroglyphidae
Arthropods
Complex Mixtures

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology
  • Clinical Biochemistry
  • Microbiology (medical)

Cite this

Jeong, Kyoung Yong ; Yum, Hye Yung ; Lee, In Yong ; Ree, Han Il ; Hong, Chein Soo ; Kim, Dong Soo ; Yong, Tai Soon. / Molecular Cloning and Characterization of Tropomyosin, a Major Allergen of Chironomus kiiensis, a Dominant Species of Nonbiting Midges in Korea. In: Clinical and Diagnostic Laboratory Immunology. 2004 ; Vol. 11, No. 2. pp. 320-324.
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abstract = "Chironomids are widely and abundantly distributed in the vicinity of standing waters. Larvae of Chironomus and some other genera are known to contain hemoglobins, which have been described as a major allergen, and the adults that have no hemoglobins also have been reported to contain allergens. In this study, we tried to establish the role of chironomid allergy and characterize the allergen of Chironomus kiiensis adults. Skin tests using C. kiiensis adult extracts were performed on patients with allergic symptoms. A cDNA library of C. kiiensis adults was screened with C. kiiensis immune mouse sera to identify allergens, and results were confirmed using skin test-positive human sera. Recombinant allergen was expressed in Escherichia coli and purified by affinity chromatography using nickel-nitrilotriacetic acid agarose to investigate its allergenic properties. Out of 275 allergic patients 14.2{\%} showed a positive reaction to C. kiiensis adult crude extracts in the skin test. The tropomyosin was cloned by immunoscreening and expressed in Escherichia coli. C. kiiensis tropomyosin has a high homology at the amino acid level with tropomyosins which were previously known to be allergens in various arthropods (Periplaneta americana, 86.3{\%}; Panulirus stimpson, 78.9{\%}; Dermatophagoides pteronyssinus, 76.5{\%}). Specific immunoglobulin E antibodies reacting to recombinant tropomyosin were detected in 17 (81{\%}) of 21 patients whose skin test results were positive. Cross-reactivity against house dust mites and other insects was noticed with mouse anti-recombinant tropomyosin immune serum. C. kiiensis adults were shown to be an important source of inhalant allergens in Korea. Molecular cloning of C. kiiensis tropomyosin was performed and IgE reactivity was demonstrated using skin test-positive human sera. Recombinant tropomyosin will be useful for further studies or clinical applications.",
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Molecular Cloning and Characterization of Tropomyosin, a Major Allergen of Chironomus kiiensis, a Dominant Species of Nonbiting Midges in Korea. / Jeong, Kyoung Yong; Yum, Hye Yung; Lee, In Yong; Ree, Han Il; Hong, Chein Soo; Kim, Dong Soo; Yong, Tai Soon.

In: Clinical and Diagnostic Laboratory Immunology, Vol. 11, No. 2, 01.03.2004, p. 320-324.

Research output: Contribution to journalArticle

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AU - Jeong, Kyoung Yong

AU - Yum, Hye Yung

AU - Lee, In Yong

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AU - Kim, Dong Soo

AU - Yong, Tai Soon

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