1. ResultsResults NO. 18 都市部花粉アレルゲン特性調査プロジェ クト A practical project for the characteristic investigation of pollen allergen in the urban region Project Participants: Di Wu, Nakamura Shinichi, Gong Xiumin, Morita Jun Supervisor: Seiyo OO Measurement of Japanese cedar pollen (Y2010) antigen Cry j 1 concentration by using ELISA Table 1 Cross-antigenicity detection by polyclonal antibodies Cry j 1 concentration by different particle size Fig. 7. Sampling Site location: Waste disposal vicinity in Saitama university campus Sampling Period: February - May, 2010 Sampling Purpose: For pollen counting calculation and pollen antigen measurement Sampling Information Materials and Methods The respond values by cedar are more efficient than cypress; notwithstanding, the existence of cypress antigens has been detected. Cooperators: National Institute for Environmental Studies, Waste Treatment cooperation, Center for Environmental Science in Saitama In Japan, Japanese cedar (Cryptometria japonica) and cypress (Chamaecyparis obtusa) pollens are considered to be the major unique allergens which can give rise to severe pollinosis and their extent of dispersal is quite wide, especially in the urban area. In this study, Japanese cedar and cypress pollens were collected in Saitama University campus during the airborne pollen season of 2010. Morphological analyses have been performed by an optical microscope in order to complete the pollen counts observation. Airborne Cry j 1 particles, the allergenic content of cedar pollen, were collected by using a high volume Andersen air sampler and measured by the enzyme-linked immunosorbent assay method. Allergenic measurements from the fresh Japanese cedar and cypress pollens as well as their cross- antigenicity were comparatively performed by using the surface plasmon resonance method. ObjectiveObjective IntroductionIntroduction In Japan, Japanese cedar (Cryptomeria japonica) (Fig.1) and cypress (Chamaecyparis obtusa) (Fig.2) comprise more than 46 % of forest, which probably accounts for the increasing airborne pollen counts 1). Pollinosis caused by Japanese cedar pollen is one of the commonest allergic respiratory diseases in Japan. Ceda r Cypress Fig. 2. Fig. 1. Cross-antigenicity between Japanese cedar and cypress has been examined. Durham Sampling site and various samplers Fig. 3. Stainless disk of 23 cm diameter prop of 7.6 cm in height 2.5 cm separates from the slide holder bottom board Sampling site KH3000-01 automatical pollen monitor Identification and measurement of dispersion pollen Fig. 4. ● Optical microscope Pollens Cover glass (18mmX18mm) Slide glass Fig. 7 indicates the Cry j 1 concentration which particle size above 7 mm and below 1.1 mm for every 3 days internally. It is considered that the Cry j 1 concentration should be influenced on account of the precipitation, as the daily precipitation on March 7 equaled to 10 mm as well as on March 9 (23.5 mm) and March 16 (10.5 mm). Cry j 1 concentration which particle size below 1.1 mm almost occupied completely. During the period March 23 to March 28, the Cry j 1 concentration was lower accompanied by the fewer pollen grains. On most period of February, pollen counts curve performed smoothly until February 25. Japanese cedar pollen concentrated scattering during the first-10 days of March and the pollen amount tended to increase gradually due to the Japanese cypress pollen began scattering. 2010 pollen samples counted by using Durham sampler Pollen counts Fig. 6. 423 grains/cm 2 Future Plan ◆ Correlation between pollen grain counts and pollen antigen concentration ◆ Build models for reliable pollen prediction in the Saitama City including mathematical equation establishment and simulation with the lastest data by ourselves 【 Reference 】 1) Sakurai K., Naito K., Ishii G., Ogawa, T., Takeuchi K. Miyata S. and Baba R. Influence of local antigen exposure dose in the upper respiratory tract on sensitization with cedar pollen. Allergology International, 2002, 51: 9-12. Airborne pollen counts calculation ● Due to the polyclonal antibody, anti-Cry j 1 rabbit IgG fraction, was obtained from Cry j 1 antigen immunized rabbit serum, the Response Units (RU) value (273 and 222) of cedar pollen are more efficient than cypress pollen obviously. ● Indeed, for the reaction of cypress antigen, the induction sensitivity has not reached the merit RU values (45 and 35) when the cypress extraction time is 15 minutes and implantation volume equals to 70  l. ● Moreover, it almost obtain the results approximately even though the extraction time difference for 15 minutes and 24hrs respectively, that is the reason we suggest the cypress dilute solution which was extracted by 15 minutes, volume equaled to 160  L is the best choice to precede the standard solution confirmation experiment for the next partial. Biacore J and Principle of Surface plasmon resonace Fig. 5. Measurement of cedar and cypress pollen allergens by SPR