Cloning of the American cockroach Cr-PII allergens: Evidence for the existence of cross-reactive allergens between species

Chii Huei Wu, Miau-Yaun Wang, Mey Fann Lee, Chiou Ying Y. Kao, Shue Fen Luo

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Background: Previously, we have identified the 28 and 32 kd proteins as additional important allergens from the American cockroach (Periplaneta americana) Cr-PII allergenic fraction. Objective: The aim of this study was the cloning of P. americana Cr-PII allergens. Methods: A λgt22A cDNA library constructed from P. americana mRNA was packaged into Escherichia coli Y1090 (r-), and clones recognized by murine anti-Cr-PII monoclonal antibodies and human IgE antibodies were isolated, sequenced, and subcloned into pET 21 and expressed in E. coli BL21(DE3). Results: Six Cr-PII-positive clones recognized by human IgE antibodies were isolated. Two clones, C6 and C17, were sequenced, and we found encoding proteins of 228 and 274 amino acids with no cysteine or any potential N-glycosylation site, with predicted masses of 25.8 and 31.14 kd respectively. Both molecules contain internal repeated sequences with a 94% identity between them. C6 and C17 showed 59% and 77.3% skin reactivities, respectively, on 22 cockroach-sensitive atopic patients. Both clones were found to have 28.9% to 31.8% identities to ANG12 protein, a precursor of the African malaria mosquito (Anopheles gambiae) and 82.7% to 85.1% identity to a nucleotide sequence of the German cockroach (Blattella germanica) Bla g Bd90K allergen. The anti-C6 and anti-C17 antibodies were able to recognize Cr-PII, recombinant proteins, five commercial American extracts, and two German cockroach extracts. Moreover, the binding of anti-Co and anti-C17 antibodies to recombinant protein can be inhibited by B. germanica crude extract. Furthermore, Northern blot analyses have shown that B. germanica mRNAs could be detected by both cDNA probes. Conclusion: Our findings provide the first evidence of antigenie cross-reactivity between P. americana and B. germanica allergens on molecular levels. The results will be a great aid in facilitating the epitope mapping and improving diagnostic and therapeutic reagents for both cockroach species.

Original languageEnglish
Pages (from-to)832-840
Number of pages9
JournalJournal of Allergy and Clinical Immunology
Volume101
Issue number6 II
Publication statusPublished - 1998 Dec 1

Fingerprint

Periplaneta
Allergens
Organism Cloning
Clone Cells
Blattellidae
Cockroaches
Recombinant Proteins
Immunoglobulin E
Anti-Idiotypic Antibodies
Escherichia coli
Epitope Mapping
Anopheles gambiae
Messenger RNA
Protein Precursors
Antibodies
Culicidae
Complex Mixtures
Gene Library
Glycosylation
Northern Blotting

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

@article{b1a9359267274b54b773b3b484a9192b,
title = "Cloning of the American cockroach Cr-PII allergens: Evidence for the existence of cross-reactive allergens between species",
abstract = "Background: Previously, we have identified the 28 and 32 kd proteins as additional important allergens from the American cockroach (Periplaneta americana) Cr-PII allergenic fraction. Objective: The aim of this study was the cloning of P. americana Cr-PII allergens. Methods: A λgt22A cDNA library constructed from P. americana mRNA was packaged into Escherichia coli Y1090 (r-), and clones recognized by murine anti-Cr-PII monoclonal antibodies and human IgE antibodies were isolated, sequenced, and subcloned into pET 21 and expressed in E. coli BL21(DE3). Results: Six Cr-PII-positive clones recognized by human IgE antibodies were isolated. Two clones, C6 and C17, were sequenced, and we found encoding proteins of 228 and 274 amino acids with no cysteine or any potential N-glycosylation site, with predicted masses of 25.8 and 31.14 kd respectively. Both molecules contain internal repeated sequences with a 94{\%} identity between them. C6 and C17 showed 59{\%} and 77.3{\%} skin reactivities, respectively, on 22 cockroach-sensitive atopic patients. Both clones were found to have 28.9{\%} to 31.8{\%} identities to ANG12 protein, a precursor of the African malaria mosquito (Anopheles gambiae) and 82.7{\%} to 85.1{\%} identity to a nucleotide sequence of the German cockroach (Blattella germanica) Bla g Bd90K allergen. The anti-C6 and anti-C17 antibodies were able to recognize Cr-PII, recombinant proteins, five commercial American extracts, and two German cockroach extracts. Moreover, the binding of anti-Co and anti-C17 antibodies to recombinant protein can be inhibited by B. germanica crude extract. Furthermore, Northern blot analyses have shown that B. germanica mRNAs could be detected by both cDNA probes. Conclusion: Our findings provide the first evidence of antigenie cross-reactivity between P. americana and B. germanica allergens on molecular levels. The results will be a great aid in facilitating the epitope mapping and improving diagnostic and therapeutic reagents for both cockroach species.",
author = "Wu, {Chii Huei} and Miau-Yaun Wang and Lee, {Mey Fann} and Kao, {Chiou Ying Y.} and Luo, {Shue Fen}",
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Cloning of the American cockroach Cr-PII allergens : Evidence for the existence of cross-reactive allergens between species. / Wu, Chii Huei; Wang, Miau-Yaun; Lee, Mey Fann; Kao, Chiou Ying Y.; Luo, Shue Fen.

In: Journal of Allergy and Clinical Immunology, Vol. 101, No. 6 II, 01.12.1998, p. 832-840.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cloning of the American cockroach Cr-PII allergens

T2 - Evidence for the existence of cross-reactive allergens between species

AU - Wu, Chii Huei

AU - Wang, Miau-Yaun

AU - Lee, Mey Fann

AU - Kao, Chiou Ying Y.

AU - Luo, Shue Fen

PY - 1998/12/1

Y1 - 1998/12/1

N2 - Background: Previously, we have identified the 28 and 32 kd proteins as additional important allergens from the American cockroach (Periplaneta americana) Cr-PII allergenic fraction. Objective: The aim of this study was the cloning of P. americana Cr-PII allergens. Methods: A λgt22A cDNA library constructed from P. americana mRNA was packaged into Escherichia coli Y1090 (r-), and clones recognized by murine anti-Cr-PII monoclonal antibodies and human IgE antibodies were isolated, sequenced, and subcloned into pET 21 and expressed in E. coli BL21(DE3). Results: Six Cr-PII-positive clones recognized by human IgE antibodies were isolated. Two clones, C6 and C17, were sequenced, and we found encoding proteins of 228 and 274 amino acids with no cysteine or any potential N-glycosylation site, with predicted masses of 25.8 and 31.14 kd respectively. Both molecules contain internal repeated sequences with a 94% identity between them. C6 and C17 showed 59% and 77.3% skin reactivities, respectively, on 22 cockroach-sensitive atopic patients. Both clones were found to have 28.9% to 31.8% identities to ANG12 protein, a precursor of the African malaria mosquito (Anopheles gambiae) and 82.7% to 85.1% identity to a nucleotide sequence of the German cockroach (Blattella germanica) Bla g Bd90K allergen. The anti-C6 and anti-C17 antibodies were able to recognize Cr-PII, recombinant proteins, five commercial American extracts, and two German cockroach extracts. Moreover, the binding of anti-Co and anti-C17 antibodies to recombinant protein can be inhibited by B. germanica crude extract. Furthermore, Northern blot analyses have shown that B. germanica mRNAs could be detected by both cDNA probes. Conclusion: Our findings provide the first evidence of antigenie cross-reactivity between P. americana and B. germanica allergens on molecular levels. The results will be a great aid in facilitating the epitope mapping and improving diagnostic and therapeutic reagents for both cockroach species.

AB - Background: Previously, we have identified the 28 and 32 kd proteins as additional important allergens from the American cockroach (Periplaneta americana) Cr-PII allergenic fraction. Objective: The aim of this study was the cloning of P. americana Cr-PII allergens. Methods: A λgt22A cDNA library constructed from P. americana mRNA was packaged into Escherichia coli Y1090 (r-), and clones recognized by murine anti-Cr-PII monoclonal antibodies and human IgE antibodies were isolated, sequenced, and subcloned into pET 21 and expressed in E. coli BL21(DE3). Results: Six Cr-PII-positive clones recognized by human IgE antibodies were isolated. Two clones, C6 and C17, were sequenced, and we found encoding proteins of 228 and 274 amino acids with no cysteine or any potential N-glycosylation site, with predicted masses of 25.8 and 31.14 kd respectively. Both molecules contain internal repeated sequences with a 94% identity between them. C6 and C17 showed 59% and 77.3% skin reactivities, respectively, on 22 cockroach-sensitive atopic patients. Both clones were found to have 28.9% to 31.8% identities to ANG12 protein, a precursor of the African malaria mosquito (Anopheles gambiae) and 82.7% to 85.1% identity to a nucleotide sequence of the German cockroach (Blattella germanica) Bla g Bd90K allergen. The anti-C6 and anti-C17 antibodies were able to recognize Cr-PII, recombinant proteins, five commercial American extracts, and two German cockroach extracts. Moreover, the binding of anti-Co and anti-C17 antibodies to recombinant protein can be inhibited by B. germanica crude extract. Furthermore, Northern blot analyses have shown that B. germanica mRNAs could be detected by both cDNA probes. Conclusion: Our findings provide the first evidence of antigenie cross-reactivity between P. americana and B. germanica allergens on molecular levels. The results will be a great aid in facilitating the epitope mapping and improving diagnostic and therapeutic reagents for both cockroach species.

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