Allergenic Proteins of Mediterranean Cypress (Cupressus sempervirens L.) and Sabin Juniper (Juniperus sabina L.) Pollens in Istanbul

Thesis Type: Postgraduate

Institution Of The Thesis: Istanbul University, Faculty Of Science, Moleküler Biyoloji ve Genetik Bölümü, Turkey

Approval Date: 2015

Thesis Language: Turkish

Student: Şahin Taş

Principal Supervisor (For Co-Supervisor Theses): Emine Şeküre Nazlı Arda

Co-Supervisor: Aslı Gelincik


Wind-mediated spreading pollens are important aeroallergens. When they are released to atmosphere in sufficient amount, they can cause the development of diseases such as asthma, rhinitis, conjunctivitis etc. in allergically hypersensitive individuals. The spreading of pollens are affected by their amount in the air, their structures, geographic areas and the climate. Therefore, the amount of pollens that hypersensitive individuals who live in different regions are exposed to may differ and different allergic reactions may ocur in effected individuals. The pollens of cypress family (Cupressaceae) trees are potential aeroallergens. Among them especially the pollens of cypress (Cupressus) and junipers (Juniperus) are the most important allergen sources. Cypress species produce high amount of pollens. The high amount of pollen production enhance the pollen sensitivity. Some species in cypress family are widely and naturally distributed in Turkey. Thus the research on region-spesific allergenic effects of these species becomes important. In this thesis project the allergenic proteins of pollens from Mediterranean cypress (Cupressus sempervirens L.) and Sabin juniper (Juniperus sabina L.) in cypress family (Cupressaceae) deployed in Istanbul were investigated. Pollens were collected in pollination period and their extracts were prepared. The pollen extracts were diluted in certain ratios and were subjected to patients with allergic rhinits and to healthy individuals without any allergic reactions (control group) via skin prick and nasal provacation tests. After dissolving the pollen extracts in water, proteins were separated by sodium dodecyl sulphate polyacrilamide gel electrophoresis (SDS-PAGE) according to their molecular weights. The separated proteins were transferred to polyvinylidene fluoride (PVDF) membrane by Western blotting and were treated with patient serums (primary antibody) for inducing the reaction between IgE antibodies and allergenic proteins. Subsequently the membrane was treated with secondary antibody (horseradish peroxidase conjugated mouse anti-human IgE antibody). The observed bands were considered as allergenic proteins and were evaluated with other clinical data. It was observed that spesific IgE antibodies bound to total 8 pollen proteins betwen ~20-68 kDa in 5 out of 18 patients implemented Mediterranean cypress pollen extract, and implemented Sabine juniper pollen extract bound to total 9 pollen proteins between ~22-85 kDa. spesific IgE antibodies in 9 out of 17 patients. No band was detected in Western blotting studies performed with control individuals.