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Mango profilin: cloning, expression and cross-reactivity with birch pollen profilin Bet v 2
Juanjuan Song1, Hongyun Zhang, Zhigang Liu
1College of life science, Shenzhen University, Shenzhen, 518060, China.
Abstract:
Mango can cause severe anaphylactic reactions. Profilin has been assumed partly responsible for the cross-reactivity between mango fruit and other allergens but has not been finally clarified. In this study, two isoforms of mango fruits profilin were amplified by RT-PCR and 3'RACE from total RNA. Each mango profilin cDNA includes an open reading frame coding for 131 amino acids. The deduced amino acid sequence of the corresponding protein show high identity with other allergenic profilins. Expression of the recombinant mango profilin was carried out in Escherichia coli BL21(DE3) using vector PET28a and the purification of the recombinant protein was performed via affinity chromatography with Ni+ coupled to sepharose. IgE reactivity of recombinant mango profilin was investigated by immunoblot and 8 of 18 mango-allergic patients tested presented specific IgE-antibodies to recombinant mango profilin. IgE-inhibition and ELISA inhibition experiments were performed to analyze mango profilin cross-reactivity with profilins from birch pollen and high cross-reactivities have been found.
Insights
Mango profilin, a key allergen, triggers allergic reactions and shows high cross-reactivity with birch pollen profilin. This study clarifies its role in mango allergy, identifying specific IgE antibodies in patients.
Area of Science:
- Immunology
- Allergology
- Molecular Biology
Background:
- Mango allergy can lead to severe anaphylaxis.
- Profilins are suspected allergens responsible for cross-reactivity between mango and other sources, but their role remains unclear.
Purpose of the Study:
- To investigate the role of mango profilins in allergic reactions.
- To characterize the cross-reactivity of mango profilin with other profilins.
Main Methods:
- Amplification of two mango profilin isoforms using RT-PCR and 3'RACE.
- Expression and purification of recombinant mango profilin in E. coli.
- Investigation of IgE reactivity using immunoblotting, IgE-inhibition, and ELISA inhibition assays.
Main Results:
- Mango profilin cDNA codes for a 131-amino acid protein with high identity to other allergenic profilins.
- Recombinant mango profilin showed specific IgE binding in 8 out of 18 mango-allergic patients.
- High cross-reactivity was observed between mango profilin and birch pollen profilin.
Conclusions:
- Mango profilin is a significant allergen involved in mango-induced allergic reactions.
- Mango profilin exhibits substantial cross-reactivity with profilins from other allergens like birch pollen.
