Reduction of Allergenicity of Litchi chinensis Flowers Pollen Protein Conjugated with Polysaccharide by Maillard Reaction

  • Ranajit Kumar Shaha Department of Agro-Industry, Faculty of Agro-Industry & Natural Resources, University Malaysia Kelantan, Jel Campus, 17600 Jeli, Kelantan, Malaysia.
  • Nitai Roy Department of Biochemistry & Molecular Biology, University of Rajshahi, Rajshahi-6205, Bangladesh.
  • G. Talukdar Department of Biochemistry & Molecular Biology, University of Rajshahi, Rajshahi-6205, Bangladesh.
  • I. Hossain Department of Biochemistry & Molecular Biology, University of Rajshahi, Rajshahi-6205, Bangladesh.
  • M. Shariar Shovon Department of Biochemistry & Molecular Biology, University of Rajshahi, Rajshahi-6205, Bangladesh.
  • N. K. Sana Department of Biochemistry & Molecular Biology, University of Rajshahi, Rajshahi-6205, Bangladesh.
  • S. R. Kabir Department of Biochemistry & Molecular Biology, University of Rajshahi, Rajshahi-6205, Bangladesh.
  • K. K. Biswas Department of Biochemistry & Molecular Biology, University of Rajshahi, Rajshahi-6205, Bangladesh.
Keywords: Litchi chinensis (Litchi), sensitization, Swiss albino mice, Eosinophils, Neutrophils

Abstract

Background: Allergy to pollen from gymnosperms is well documented in the west. The objective was to define the allergologic protein from Litchi chinensis (Litchi) pollen and conjugate the protein with polysaccharides by Maillard reaction to reduce the allergic effect of that protein.

Methods: Total soluble proteins were extracted from the pollen of Litchi flower pollen and subjected to ammonium sulphate precipitation at 80% saturation. Pollen antigen from Litchi chinensis (Litchi) was prepared by gel cutting method and characterized by biochemical and designated by LFPP. The homogeneity of this protein was demonstrated by a single band on SDS-PAGE. The protein then conjugated with galactomannan through Maillard Reaction. The resulting purified pollen protein and conjugated protein were administered to the Swiss albino mice as amount of 5.8mg/kg body weight.

Results: The total protein was then separated on a 12% SDS-Polyacrylamide gel which revealed 5 bands between molecular weight range of 29kDa and 69kDa. Each band was recovered from the gel by electroelution and sent for skin tests. 28kDa proteins was the only allergenic protein while others were not shown reactivity in patients. Intraperitoneal injection of the purified protein (LFPP) caused a significant rise in the levels of neutrophils (38-81%) and eosinophils (3-14%) compared to control (P<0.001) whereas conjugated protein caused only a 2% increase of both neutrophils and eosinophils level. On the other hand treatment with LFPP-galactomannan conjugate causes no such change in physical appearance with eosinophils and neutrophils level.

Conclusion: The present study demonstrates that the protein extracted and purified from Litchi flowers pollen has been recognized as a new allergen from Bangladesh for the first time and the allergic effects can be reduced by conjugation with polysaccharides.

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References

[1]. D'Amato, G., Spieksma, F.T., Liccardi, G., Jäger, S., Russo, M., Kontou-Fili, K., Nikkels, H., Wüthrich, B., Bonini, S. (1998). Pollen-related allergy in Europe. Allergy, 53: 567-578.
[2]. Goldberg, A., Confino-Cohen, R., Waisel, Y. (1998). Allergic response to pollen of ornamental plants: high incidence in general atopic population and especially among flower growers. J. Allergy Clin. Immunol., 102: 210-214.
[3]. Afferni, C., Iacovacci, P., Barletta, B., Di Felice, G., Tinghino, R., Mari, A., Pini, C. (1999). Role of carbohydrate moieties in IgE binding to allergenic components of Cupressus arizonica pollen extract. Clin. Exp. Allergy, 29:1087-1094.
[4]. Bapat, B.N. & Bapat, H.B., (1994). The patient of atopy: a more complete approach. In: Recent Trends in Aerobiology, Allergy and Immunology (ed. S.N. Agashe), pp-183-195. Oxford & IBH Publ. Co. Pvt. Ltd. New Delhi.
[5]. Babiker, E., Fadil, E., Hiroyuki, A., Matsudomi, N., Iwata, H., Ogawa, T., Bando, N., & Kato, A. (1998). Effect of polysaccharides conjugation or transglutaminase treatment on the allergenicity and functional properties of soy protein. J. Agric. Food Chem., 46: 866-871.
[6]. Chanda, S. (1994). Pollen grains as aeroallergens: morphological, biological and chemical approach. In: Agashe SN (Ed): Recent Trends in Aerobiology, Allergy and Applied Immunology, 85-92. Oxford and IBH Publ., New Delhi 1994.
[7]. Cimignoli, E., Broccucci, L., Cernetti, C., Gerli, R., Spinozzi, F. (1992). Isolation and partial characterization of Cupressus sempervirens allergens. Aerobiologia, 8:465-470.
[8]. Fujiwara, H., Masuda, M., Kubo, N., Okochi, H., Shimoshikiryo, K., Okamoto, H., Hara, K., Ogawa, N., Takahashi, H. (1997). Seasonal changes of laboratory data in patients with Japanese cedar pollinosis. Rinsho Byori, 45(5): 487-92.
[9]. Grafin, D.E. (1990). Isoelectric focusing. Methods Enzymology. 182: 459-477.
[10]. G. Talukdar., I. Hossain., Nitai, C. Roy., N.K. Sana., S.R. Kabir., K.K. Biswas., N. Roy and Ranajit Kumar Shaha (2012). Allergen Protein from Mango (Mangifera indica) Flowers Pollen. AsPac. J. Mol. Biol. Biotechnology, 20: 11-18.
[11]. Humbles, A.A., C.M. Lloyd., S.J. McMillan., D.S. Friend., G. Xanthou., E.E. McKenna., S. Ghiran., N.P. Gerard., C. Yu, S.H. Orkin., C. Gerard (2004). A critical role for eosinophils in allergic airways remodeling. Science, 305:1776-1779.
[12]. Lee, J.J., Dimina, D., Macias, M.P., Ochkur, S.I., McGarry, M.P., O'Neill, K.R., Protheroe, C., Pero, R., Nguyen, T., Cormier, S.A., Lenkiewicz, E., Colbert, D., Rinaldi, L., Ackerman, S.J., Irvin, C.G., Lee, N.A. (2004). Defining a link with asthma in mice congenitally deficient in eosinophils. Science, 305:1773-1776.
[13]. Little, S.A., MacLeod, K.J., Chalmers, G.W., Love, J.G., McSharry, C. and Thomson, N.C. (2002). Association of forced expiratory volume with disease duration and sputum neutrophils in chronic asthma. American Journal of Medicine, 112: 446-452.
[14]. Lowry, O.H., Roseborough, N.J., Farr, A.L. and Randall, R.J. (1951). Protein measurement with the Folin phenol reagent. J. Boil. Chem., 193: 265-273.
[15]. Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophages T4. Nature, 277:680-685.
[16]. Iacovacci, P., Afferni, C., Barletta, B., Tinghino, R., Di Felice, G., Pini, C., Mari, A. (1998). Juniperus oxycedrus: a new allergenic pollen from Cupressaceae family. J. Allergy Clin. Immunol., 101:755-761.
[17]. Usui, M., Saito, A., Taniguchi, N., Nishijima, N., Azakami, H., Kato, A. (2003). Reduction of antigenicity of Cry j I, Major allergen of Japanese ceder pollen, by the attachment of polysaccharides. Biosci. Biotechnol. Biochem., 67(11): 2425-2430.
[18]. Mondal, AK. & Mandal, S. (1997). Biochemical and clinical studies of Ailanthus excels Roxb. Pollen with reference to its dispersal. Bangladesh J. Botany., 26(2): 115-120.
[19]. Mondal, A.K., Parui, S., Biswas, S.R. and Mondal, S. (1997a). Identification of the allergenic proteins of Ipomea fistulosa pollen- partial characterization and sensitivity test. Grana, 36:301-305.
[20]. Mondal, A.K., Parui, S. and Mondal, S. (1997b). Aerobiology and some chemical parameters of the pollen of a few members of Mimosaceae in Burdwan District, West Bengal. Res. J. Chem. Environ., 1:12-19.
[21]. Mondal, A.K., Parui, S. and Mondal, S. (1998). Biochemical analysis of four species of Cassia L. Pollen. Aerobiologia, 14:45-50.
[22]. Newmark, F.M. and Itkin, I.H. (1967). Asthma due to pine pollen. J. Allergy, 25:251-252.
[23]. García-Ortega, P., Martínez, J., Martínez, A., Palacios, R., Belmonte, J., Richart, C. (1992). Mercurialis annua pollen: a new source of allergic sensitization and respiratory disease. J. Allergy Clin. Immunol., 89: 987-993.
[24]. Pasha, M.K. and Mohammad Sohrab Hossain, (2009). Airborne pollen grains at Chittagong University campus, Bangladesh. Bangladesh J. Bot., 38(1): 39-46.
[25]. Pettyjohn, M.E., Levetin, E. (1997). A comparative biochemical study of conifer pollen allergens. Aerobiologia, 13:259-267.
[26]. Parui, S. & Mandal, S. (1998). Biochemical analysis and skin sensitivity test of the allergenic pollen of Datura metel L. Curr. Sci., 74(1): 66-68.
[27]. Parui, S., Mondal, A.K and Mondal, S. (1999). Identification and partial characterization of the allergenic proteins of Ricinus communis L. Pollen –a new approach. Grana, 38(5):311-315.
[28]. Rudramuniyappa, C.K. (1996). Pollen wall is a living system: an analysis. -In: Pollen-spore research emerging strategies (ed. C.P. Malik) pp-67-94. –Adv. Pollen Spore Res. 21. Today & Tomorrow’s Print & Publ. New Delhi.
[29]. Singh, A.B. and Singh, A. (1994). Pollen allergy- a global scenario. In: Recent Trends in Aerobiology, Allergy and Immunology. Agashe, S. N. Eds. Oxford and IBH, 143-173.
[30]. Shivpuri, D.N., Singh A.B., Babu, C.E. (1979). New allergenic pollens of Delhi state, India and their clinical significance. Ann. Allergy, 42: 49-52.
[31]. Singh, A.B. (1987). Airborne pollen types of allergenic significance in India. Adv. Aerobiol., (Experientia. Suppl.) 51: 61-64.
[32]. Singh A.B., Malik P., Parkash, D. and Gangal, S.V. (1993). Identification of specific IgE binding proteins in castor bean (Ricinus communis) pollen obtained from different source materials. Grana., 32: 376-380.
[33]. Salvatore Feo, Roberta Cocchiara and Domenico Geraci (1984). Allergens of Parietaria judaica Pollen-I. Purification and characterization of a hapten and a low molecular weight allergenic Peptide. Mol. Immunol., 21: 25-36.
[34]. Wenzel, S.E., S.J. Szefler., D.Y. Leung., S.I. Sloan., M.D. Rex., R.J. Martin (1997). Bronchoscopic evaluation of severe asthma: persistent inflammation associated with high dose glucocorticoids. Am. J. Respir. Crit. Care Med., 156:737-743.
[35]. Wilson, K. & Goulding, K.H. (1986). A Biologist’s guide to principles and techniques of Practical Biochemistry. E. Arnold Ltd., London.
Published
2012-04-01
How to Cite
Shaha, R., Roy, N., Talukdar, G., Hossain, I., Shovon, M., Sana, N., Kabir, S., & Biswas, K. (2012). Reduction of Allergenicity of Litchi chinensis Flowers Pollen Protein Conjugated with Polysaccharide by Maillard Reaction. Journal of Advanced Laboratory Research in Biology, 3(2), 90-98. Retrieved from http://e-journal.sospublication.co.in/index.php/jalrb/article/view/105
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