ResultsResultsResultsResults

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 regionA practical project for the characteristic investigation of pollen allergen in the urban region

Project Participants: Di Wu, Nakamura Shinichi, Gong Xiumin, Morita JunSupervisor: Seiyo OO

Measurement of Japanese cedar pollen (Y2010) antigen Cry j 1 concentration by using ELISA

Sample Extraction time Volume Number of times RU1 105.42 112.33 125.41 44.72 351 132.82 127.13 123.61 272.82 221.6

70ml

160ml

160ml

15 mins 160ml

Cedar 24 hrs

Cypress 15 mins

24 hrs

0.00

0.05

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0.15

0.20

3/5～3/7 3/8～3/10 3/11～3/13 3/14～3/16 3/17～3/19 3/20～3/22 3/23～3/25 3/26～3/28

Date

Cry

j 1

con

cent

rati

on(n

g/m

3)

>7.0 3.3-7.0 <1.1 μm

0.00

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0.15

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3/5～3/7 3/8～3/10 3/11～3/13 3/14～3/16 3/17～3/19 3/20～3/22 3/23～3/25 3/26～3/28

Date

Cry

j 1

con

cent

rati

on(n

g/m

3)

>7.0 3.3-7.0 <1.1 μm

Table 1Table 1 Cross-antigenicity detection by polyclonal antibodies

Cry j 1 concentration by different particle size Fig. Fig. 7.7.

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 MethodsMaterials and MethodsMaterials and MethodsMaterials 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.

ObjectiveObjectiveObjectiveObjective

IntroductionIntroductionIntroductionIntroduction

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 counts1). Pollinosis caused by Japanese cedar pollen is one of the commonest allergic respiratory diseases in Japan.

Cedar

CypressFig. Fig. 2. 2.

Fig. Fig. 1. 1.

Cross-antigenicity between Japanese cedar and cypress has been examined.

Durham

Sampling site and various samplersFig. 3. Fig. 3.

Stainless disk of 23 cm diameterprop 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 pollenFig. Fig. 4. 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.

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ins/

cm2 )

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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. Fig. 6.6.

423 grains/cm2

Future PlanFuture PlanFuture PlanFuture 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

【【 ReferenceReference 】】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 ml.● 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 mL is the best choice to precede the standard solution confirmation experiment for the next partial.

Injector

Flow cell

Pump

Flow channel

Antigen

Antibody

Gold thin film

Glass

Waste liquid

Prism

Light sourcePhotoreceiver

Injector

Flow cell

Pump

Flow channel

Antigen

Antibody

Gold thin film

Glass

Waste liquid

Prism

Light sourcePhotoreceiver

Biacore J and Principle of Surface plasmon resonaceFig. Fig. 5.5.

Measurement of cedar and cypress pollen allergens by SPR