Identification of Aspergillus (A flavus and A niger) Allergens and Heterogeneity of Allergic Patients’ IgE Response
Abstract
Aspergillus species (A flavus and A niger) are important sources of inhalant allergens. Current diagnostic modalities employ crude Aspergillus extracts which only indicate the source to which the patient has been sensitized, without identifying the number and type of allergens in crude extracts. We report a study on the identification of major and minor allergens of the two common airborne Aspergillus species and heterogeneity of patients’ IgE response to them.Skin prick tests were performed on 300 patients of bronchial asthma and/or allergic rhinitis and 20 healthy volunteers. Allergen specific IgE in patients’ sera was estimated by enzyme allergosorbent test (EAST). Immunoblots were performed to identify major/minor allergens of Aspergillus extracts and to study heterogeneity of patients’ IgE response to them.Positive cutaneous responses were observed in 17% and 14.7% of patients with A flavus and A niger extracts, respectively. Corresponding EAST positivity was 69.2% and 68.7%. In immunoblots, 5 allergenic proteins were identified in A niger extract, major allergens being 49, 55.4 and 81.5 kDa. Twelve proteins bound patients’ IgE in A flavus extract, three being major allergens (13.3, 34 and 37 kDa). The position and slopes of EAST binding and inhibition curves obtained with individual sera varied from patient to patient. The number and molecular weight of IgE-binding proteins in both the Aspergillus extracts varied among patients.These results gave evidence of heterogeneity of patients’ IgE response to major/minor Aspergillus allergens. This approach will be helpful to identify disease eliciting molecules in the individual patients (component resolved diagnosis) and may improve allergen-specific immunotherapy.
1. Global Initiative for Asthma. Global strategy for asthma management and prevention. 2012 [updated 2012; accessed 2014 Mar 6]. Available from URL: http://www.ginasthma.org/guidelines-gina-report-global- strategy-for-asthma.html.
2. Bousquet J, Khaltaev N, Cruz AA, Denburg J, Fokkens WJ, Togias A, et al. Allergic Rhinitis and its Impact on Asthma (ARIA) 2008 update (in collaboration with the World Health Organization, GA(2)LEN and AllerGen). Allergy 2008; 63(Suppl 86):8-160.
3. Koppelman GH. Gene-environment interaction in allergic disease: More questions, more answers? J Allergy Clin Immunol 2007; 120(6):1266-8.
4. Simon-Nobbe B, Denk U, Pöll V, Rid R, Breitenbach M.The spectrum of fungal allergy. Int Arch Allergy Immunol 2008; 145(1):58-86.
5. Zureik M, Neukirch C, Leynaert B, Liard R, Bousquet J,Neukirch F, et al. Sensitization to airborne moulds and severity of asthma: cross sectional study from European Community respiratory health survey. BMJ 2002;325(7361):411-4.
6. Fang Z, Ouyang Z, Hu L, Wang X, Zheng H, Lin X.Culturable airborne fungi in outdoor environments in Beijing, China. Sci Total Environ 2005; 350(1-3):47-58.
7. Klaric M, Pepeljnjak S. A year-round aeromycological study in Zagreb area, Croatia. Ann Agric Environ Med 2006; 13(1):55-64.
8. Wu YH, Chan CC, Rao CY, Lee CT, Hsu HH, Chiu YH, et al. Characteristics, determinants and spatial variations of ambient fungal levels in the subtropical Taipei metropolis. Atmos Environ 2007; 41:2500-9.
9. O’Gorman CM, Fuller HT. Prevalence of culturable airborne spores of selected allergenic and pathogenic fungi in outdoor air. Atmos Environ 2008; 42:4355-68.
10. Sharma D, Dutta BK, Singh AB, Shome BR.Aerobiological, biochemical and immunological studies on some of the dominant Aspergillus species of South Assam (India). Aerobiologia 2007; 23:201-10.
11. Hedayati MT, Mayahi S, Denning DW. A study on Aspergillus species in houses of asthmatic patients from Sari City, Iran and a brief review of the health effects of exposure to indoor Aspergillus. Environ Monit Assess 2010; 168(1-4):481-7.
12. D’Amato G, Chatzrgeorgiou G, Corsico R, Gioulekas D, Jager L, Jager S, et al. Evaluation of the prevalence of skin prick test positivity to Alternaria and Cladosporium in patients with suspected respiratory allergy. A European multicenter study promoted by the subcommittee on aerobiological and environmental aspects of inhalant allergens of the European Academy of Allergology and Clinical Immunology. Allergy 1997; 52(7):711-6.
13. Ezeamuzie CI, Al-Ali S, Khan M, Hijazi Z, Dowaisan A, Thomson MS, et al. IgE mediated sensitization to mould allergens among patients with allergic respiratory diseases in a desert environment. Int Arch Allergy Immunol 2000; 121(4):300-7.
14. Mari A, Schneider P, Wally V, Breitenbach M, Simon- Nobbe B. Sensitization to fungi: Epidemiology, comparative skin tests, and IgE reactivity of fungal extracts. Clin Exp Allergy 2003; 33(10):1429-38.
15. Gioulekas D, Damialis A, Papakosta D, Spieksma FTM, Gioulekas P, Patakas D. Allergenic fungi spore records (15 years) and sensitization in patients with respiratory allergy in Thessaloniki-Greece. J Investig Allergol Clin Immunol 2004; 14(3):225-31.
16. Crameri R, Garbani M, Rhyner C, Huitema C. Fungi: the neglected allergenic sources. Allergy 2014; 69(2):176-85.
17. Chapman MD, Ferreira F, Villalba M, Cromwell O, Bryan D, Becker WM, et al. The European Union CREATE project: A model for international standardization of allergy diagnostics and vaccines. J Allergy Clin Immunol 2008; 122(5):882-9.
18. Aggarwal S, Chhabra SK, Saxena RK, Agarwal MK.Heterogeneity of immune responses to various Aspergillus species in patients with allergic respiratory diseases. Indian J Chest Dis Allied Sci 2000; 42(4):249-58.
19. Bhalla PL, Singh MB. Biotechnology based allergy diagnosis and vaccination. Trends Biotech 2008;26(3):153-61.
20. Horner WE, Helbling A, Salvaggio JE, Lehrer SB. Fungal allergens. Clin Microbiol Rev 1995; 8(2):161-79.
21. Kurup VP, Banerjee B. Fungal allergens and peptide epitopes. Peptides 2000; 21(4):589-99.
22. Crameri R, Zeller S, Glaser AG, Vilhelmsson M, Rhyner C. Cross-reactivity among fungal allergens: A clinically relevant phenomenon? Mycoses 2009; 52(2):99-106.
23. Schwartz HJ, Citron KM, Chester EH, Kaimal J, Barlow PB, Baum GL, et al. A comparison of prevalence of sensitization to Aspergillus antigens among asthmatics in Cleveland and London. J Allergy Clin Immunol 1978;62(1):9-14.
24. Maurya V, Gugnani SC, Sarma PU, Madan T, Shah A.Sensitization to Aspergillus antigens and occurrence of allergic bronchopulmonary aspergillosis in patients with asthma. Chest 2005; 127(4):1252-9.
25. Vermani M, Vijayan VK, Menon BK, Kausar MA, Agarwal MK. Physico-chemical and clinico-immunologic studies on the allergenic significance of Aspergillus tamarii, a common airborne fungus. Immunobiology 2011; 216(3):393-401.
26. Vermani M, Vijayan VK, Kausar MA, Agarwal MK.Quantification of airborne Aspergillus allergens: Redefining the approach. J Asthma 2010; 47(7):754-61.
27. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193(1):265-75.
28. Scott TA, Melvin EH. Determination of dextran with anthrone. Anal Chem 1953; 25:1656-61.
29. Laemmli UK. Cleavage of structural proteins using the assembly of the head of bacteriophage T4. Nature (Lond)1970; 227(5259):680-5.
30. Kilinc I, Alper S. Adverse effects of antihistamines on skin tests and antihistamines in pregnancy. Curr Med Chem-Anti-Inflammatory & Anti-Allergy Agents 2005;4:517-9.
31. Johnstone A, Thorpe R. Immunochemistry in practice.2nd ed. Oxford, UK: Blackwell Scientific Publications,1987:257-60.
32. Bernstein IL, Li JT, Bernstein, DI, Hamilton R, Spector S, Tan R, et al. Allergy diagnostic testing: An updated practice parameter. Ann Allergy Asthma Immunol 2008;100 (3 Suppl 3):S1-148.
33. Towbin H, Staehelin T, Gorden J. Electrophoretic transfer of proteins from polyacrylamide gel to nitrocellulose sheet: Procedures and some applications. Proc Natl Acad Sci 1979; 76(9):4350-4.
34. Prasad R, Verma SK, Dua R, Kant S, Kushwaha R, Agarwal SP. A study of skin sensitivity to various allergens by skin prick test in patients of nasobronchial allergy. Lung India 2009; 26(3):70-3.
35. Yu CJ, Chiou SH, Lai WY, Chiang BL, Chow LP.Characterization of a novel allergen, a major IgE-binding protein from Aspergillus flavus, as an alkaline serine protease. Biochem Bioph Res Comm 1999; 261(3):669-75.
36. Li CS, Hsu LY, Chou CC, Hsieh KH. Fungus allergens inside and outside the residences of atopic and control children. Arch Environ Health 1995; 50(1):38-43.
37. Chou H, Lin W, Tam MF, Wang SR, Han SH, Shen HD.Alkaline serine proteinase is a major allergen of Aspergillus flavus- a prevalent airborne Aspergillus species in the Taipei area. Int Arch Allergy Immunol1999; 119(4):282-90.
38. Shen HD, Lin WL, Tam MF, Chou H, Wang CW, Tsai JJ, et al. Identification of vacuolar serine proteinase as a major allergen of Aspergillus fumigatus by immunoblotting and N-terminal amino acid sequence analysis. Clin Exp Allergy 2001; 31(2):295-302.
39. Shen HD, Tam MF, Chou H, Han SH. The importance of serine proteinases as aeroallergens associated with asthma. Int Arch Allergy Immunol 1999; 119(4):259-64.
40. Gautam P, Sundaram CS, Madan T, Gade WN, Shah A, Sirdeshmukh R, et al. Identification of novel allergens of Aspergillus fumigatus using immunoproteomics approach. Clin Exp Allergy 2007; 37(8):1239-49.
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Issue | Vol 14, No 4 (2015) | |
Section | Original Article(s) | |
Keywords | ||
Allergens Aspergillus flavus Aspergillus niger Heterogeneity IgE response Immunoblotting |
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