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The transaldolase, a novel allergen of Fusarium proliferatum, demonstrates IgE cross-reactivity with its human analogue.

Chou H, Wu KG, Yeh CC, Tai HY, Tam MF, Chen YS, Shen HD - PLoS ONE (2014)

Bottom Line: The purified recombinant F. proliferatum transaldolase can inhibit the IgE-binding against the 37.5 kDa component of F. proliferatum and the transaldolase allergen from Cladosporium cladosporioides.More importantly, the recombinant F. proliferatum transaldolase can inhibit IgE-binding against human transaldolase in a concentration-dependent manner.Thus, a novel and important F. proliferatum transaldolase allergen was identified.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Research, Taipei Veterans General Hospital, Taipei, Taiwan, R.O.C.

ABSTRACT
Fusarium species are among airborne fungi and recognized as causative agents of human atopic disorders. However, Fusarium allergens have not been well characterized and the lack of information limits clinical diagnosis and treatment of fungal allergy. The purpose of this study is to identify and characterize important allergens of F. proliferatum. IgE-reacting F. proliferatum components were identified by immunoblot using serum samples from patients of respiratory atopic diseases. Characterization of allergens and determination of IgE cross-reactivity were performed by cDNA cloning, then homologous expression and immunoblot inhibition studies. We identified nine different F. proliferatum components that can be recognized by IgE antibodies in 17 (28%) of the 60 atopic sera tested. Components with molecular masses of about 43, 37.5 and 36.5 kDa with IgE-binding frequencies of about 88, 47 and 53%, respectively, were considered as important allergens of F. proliferatum. The 37.5 kDa IgE-binding component was putatively considered as a transaldolase protein of F. proliferatum. The full-length cDNA of F. proliferatum transaldolase was subsequently cloned. It encodes an open reading frame of 312 amino acids and has sequence identifies of 73 and 61%, respectively, with Cladosporium and human transaldolases. The purified recombinant F. proliferatum transaldolase can inhibit the IgE-binding against the 37.5 kDa component of F. proliferatum and the transaldolase allergen from Cladosporium cladosporioides. More importantly, the recombinant F. proliferatum transaldolase can inhibit IgE-binding against human transaldolase in a concentration-dependent manner. Thus, a novel and important F. proliferatum transaldolase allergen was identified. In addition to IgE cross-reactivity between the Fusarium and the Cladosporium transaldolase allergens, IgE cross-reactivity between the Fusarium and the human transaldolases also exists and might contribute to atopic manifestations in the absence of exogenous allergen exposure.

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Antigenicity of recombinant F. proliferatum, C. cladosporioides and human transaldolases.(A) Coomassie blue-stained protein profile of rFus p 4.0101, rCla c 14.0101 and recombinant human transaldolase on PVDF membranes. (B) IgE immunoblot reactivities of these three recombinant proteins analyzed by using serum samples nos. 1–3 from Fig. 1 (serum nos. 1–3) and nine serum samples from asthmatic patients (serum nos. 21–29) who showed previously IgE-binding reactvities against rCla c 14.0101 or recombinant human transaldolase. Sera from a non-atopic healthy individual (serum no. 18) and two house dust mite-sensitized atopic individuals (serum nos. 19 and 20) were included as controls.
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pone-0103488-g003: Antigenicity of recombinant F. proliferatum, C. cladosporioides and human transaldolases.(A) Coomassie blue-stained protein profile of rFus p 4.0101, rCla c 14.0101 and recombinant human transaldolase on PVDF membranes. (B) IgE immunoblot reactivities of these three recombinant proteins analyzed by using serum samples nos. 1–3 from Fig. 1 (serum nos. 1–3) and nine serum samples from asthmatic patients (serum nos. 21–29) who showed previously IgE-binding reactvities against rCla c 14.0101 or recombinant human transaldolase. Sera from a non-atopic healthy individual (serum no. 18) and two house dust mite-sensitized atopic individuals (serum nos. 19 and 20) were included as controls.

Mentions: In this study, rFus p 4.0101 was expressed as N-terminal His6-tag proteins in E.coli and purified (Fig. 3, panel A). It has an apparent molecular mass of about 38 kDa upon SDS-PAGE analysis (data not shown). The Coomassie blue-stained protein profiles of rFus p 4.0101, rCla c 14.0101 and a recombinant human transaldolase obtained commercially are shown in Fig. 3, panel A. Serum sample nos. 1–3 from Fig. 1, panel B showed positive IgE-binding against rFus p 4.0101. These three serum samples have negative IgE-binding to rCla c 14.0101 and serum no. 2 showed positive IgE-binding against human transaldolase (Fig. 3, panel B). In addition, nine serum samples from asthmatic patients with IgE-binding against rCla c 14.0101 or recombinant human transaldolase [19] were included in this study. The IgE immunoblot reactivities of these sera (serum nos. 21–29) against these three different recombinant transaldolases are shown in Fig. 3, panel B. Among these nine serum samples, eight (serum nos. 21–26, 28 and 29) showed positive IgE-binding against rFus p 4.0101. Eight (serum nos. 21–28) of these nine serum samples showed IgE-binding against rCla c 14.0101. Furthermore, serum nos. 22, 24, 26, 27 and 29 have also IgE reactivity against recombinant human transaldolase. Serum from a non-atopic individual (serum no. 18) and from two house dust mite-sensitized atopic individuals (serum nos. 19 and 20) were used as controls and showed negative IgE immunoblot reactivity against all three recombinant transaldolases (Fig. 3, panel B).


The transaldolase, a novel allergen of Fusarium proliferatum, demonstrates IgE cross-reactivity with its human analogue.

Chou H, Wu KG, Yeh CC, Tai HY, Tam MF, Chen YS, Shen HD - PLoS ONE (2014)

Antigenicity of recombinant F. proliferatum, C. cladosporioides and human transaldolases.(A) Coomassie blue-stained protein profile of rFus p 4.0101, rCla c 14.0101 and recombinant human transaldolase on PVDF membranes. (B) IgE immunoblot reactivities of these three recombinant proteins analyzed by using serum samples nos. 1–3 from Fig. 1 (serum nos. 1–3) and nine serum samples from asthmatic patients (serum nos. 21–29) who showed previously IgE-binding reactvities against rCla c 14.0101 or recombinant human transaldolase. Sera from a non-atopic healthy individual (serum no. 18) and two house dust mite-sensitized atopic individuals (serum nos. 19 and 20) were included as controls.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4116196&req=5

pone-0103488-g003: Antigenicity of recombinant F. proliferatum, C. cladosporioides and human transaldolases.(A) Coomassie blue-stained protein profile of rFus p 4.0101, rCla c 14.0101 and recombinant human transaldolase on PVDF membranes. (B) IgE immunoblot reactivities of these three recombinant proteins analyzed by using serum samples nos. 1–3 from Fig. 1 (serum nos. 1–3) and nine serum samples from asthmatic patients (serum nos. 21–29) who showed previously IgE-binding reactvities against rCla c 14.0101 or recombinant human transaldolase. Sera from a non-atopic healthy individual (serum no. 18) and two house dust mite-sensitized atopic individuals (serum nos. 19 and 20) were included as controls.
Mentions: In this study, rFus p 4.0101 was expressed as N-terminal His6-tag proteins in E.coli and purified (Fig. 3, panel A). It has an apparent molecular mass of about 38 kDa upon SDS-PAGE analysis (data not shown). The Coomassie blue-stained protein profiles of rFus p 4.0101, rCla c 14.0101 and a recombinant human transaldolase obtained commercially are shown in Fig. 3, panel A. Serum sample nos. 1–3 from Fig. 1, panel B showed positive IgE-binding against rFus p 4.0101. These three serum samples have negative IgE-binding to rCla c 14.0101 and serum no. 2 showed positive IgE-binding against human transaldolase (Fig. 3, panel B). In addition, nine serum samples from asthmatic patients with IgE-binding against rCla c 14.0101 or recombinant human transaldolase [19] were included in this study. The IgE immunoblot reactivities of these sera (serum nos. 21–29) against these three different recombinant transaldolases are shown in Fig. 3, panel B. Among these nine serum samples, eight (serum nos. 21–26, 28 and 29) showed positive IgE-binding against rFus p 4.0101. Eight (serum nos. 21–28) of these nine serum samples showed IgE-binding against rCla c 14.0101. Furthermore, serum nos. 22, 24, 26, 27 and 29 have also IgE reactivity against recombinant human transaldolase. Serum from a non-atopic individual (serum no. 18) and from two house dust mite-sensitized atopic individuals (serum nos. 19 and 20) were used as controls and showed negative IgE immunoblot reactivity against all three recombinant transaldolases (Fig. 3, panel B).

Bottom Line: The purified recombinant F. proliferatum transaldolase can inhibit the IgE-binding against the 37.5 kDa component of F. proliferatum and the transaldolase allergen from Cladosporium cladosporioides.More importantly, the recombinant F. proliferatum transaldolase can inhibit IgE-binding against human transaldolase in a concentration-dependent manner.Thus, a novel and important F. proliferatum transaldolase allergen was identified.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Research, Taipei Veterans General Hospital, Taipei, Taiwan, R.O.C.

ABSTRACT
Fusarium species are among airborne fungi and recognized as causative agents of human atopic disorders. However, Fusarium allergens have not been well characterized and the lack of information limits clinical diagnosis and treatment of fungal allergy. The purpose of this study is to identify and characterize important allergens of F. proliferatum. IgE-reacting F. proliferatum components were identified by immunoblot using serum samples from patients of respiratory atopic diseases. Characterization of allergens and determination of IgE cross-reactivity were performed by cDNA cloning, then homologous expression and immunoblot inhibition studies. We identified nine different F. proliferatum components that can be recognized by IgE antibodies in 17 (28%) of the 60 atopic sera tested. Components with molecular masses of about 43, 37.5 and 36.5 kDa with IgE-binding frequencies of about 88, 47 and 53%, respectively, were considered as important allergens of F. proliferatum. The 37.5 kDa IgE-binding component was putatively considered as a transaldolase protein of F. proliferatum. The full-length cDNA of F. proliferatum transaldolase was subsequently cloned. It encodes an open reading frame of 312 amino acids and has sequence identifies of 73 and 61%, respectively, with Cladosporium and human transaldolases. The purified recombinant F. proliferatum transaldolase can inhibit the IgE-binding against the 37.5 kDa component of F. proliferatum and the transaldolase allergen from Cladosporium cladosporioides. More importantly, the recombinant F. proliferatum transaldolase can inhibit IgE-binding against human transaldolase in a concentration-dependent manner. Thus, a novel and important F. proliferatum transaldolase allergen was identified. In addition to IgE cross-reactivity between the Fusarium and the Cladosporium transaldolase allergens, IgE cross-reactivity between the Fusarium and the human transaldolases also exists and might contribute to atopic manifestations in the absence of exogenous allergen exposure.

Show MeSH
Related in: MedlinePlus