| To understand the mechanism of mucosal immune response, indirect immunostaining with Staphylococal protein A-Horseradish peroxidase was employed for the study of distribution, change of quantity and rule of increasing or reducing of immunoglobulin A-secreting cells in Harderian Gland, providing theoretical basis for prevention and control of avian disease.In this study, chicken secretary immunoglobulin A (SIgA) was obtained by the improved method of ammonium sulfate and purified by Macro-prep High Q Anion Exchange Support ion-exchange chromatography. Polyclonal antibodies of anti-chicken SIgA was prepared. Eye-drop immunization (A and C) and intranasal Immunization (C and D) were done respectively with infectious bronchitis(IB) attenuated vaccine for one day chicken. And twice inoculation were done in the group of A and C after ten days. Results showed that concentration of purified chicken SIgA reached to 22mg/ml. It was known that the molecular weight of heavy chains, light chains and joining chains were about 64～65ku, 26～28ku and 16ku respectively from Sodium sulphate polyacrylamide get electrophoresie (SDS-PAGE). Rabbit antiserum of 1:128 was abtained from inoculated rabbits with purified chicken SIgA. The result of immunostaining indicated that SIgA positive cell displayed round or egg-like round, rarely colomn or irregular shape. SIgA cell distributed in interstitice Harderian Gland, mainly in epithelium and around gland epithelium of the duct and acini, especially mature plasma cell clung to gland epithelium. In the trachea, SIgA cell mainly distributed among tracheal mucosal epithelium and lamina propria, partial mucosa epithelia formed microcrypt, where there were more SIgA-typed plasma cell. Statistical analysis demonstrated that in the... |
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