ラットの肺洗浄液内蛋白を抗原としてウサギを免疫した.吸収操作を行って得られた抗体を使って,ホルマリン固定・パラフィン包埋されたラットの肺組織を酵素抗体法で免疫組織化学的に検索すると,形態的にクララ細胞と思われる細胞に特異的な反応を示していた.B-5固定材料ではクララ細胞のほかに細気管支上皮上のmucus blanketにも反応物質が存在した.プロテインA・ゴールドを用い電顕レベルで反応物質の局在を調べるとクララ細胞内の高電子密度の分泌顆粒に集中していた.これらの所見は抗原がクララ細胞と考えられる細胞で産生され,末梢気道に分泌されていることを示唆している.また,この抗体はラットのほかにはマウスのクララ細胞と交差反応を示したが,他の種(ハムスター,モルモット,ウサギ,犬,猫,猿,人)のそれとは反応しなかった.本報告では本抗体の作製方法について詳述した.By immunizing rabbits with the proteins in the rat lung lavage fluid, we have succeeded in producing antibodies against secretory products of the rat Clara cells. Immunohistochemically, these antibodies were demonstrated to specifically react with secretory products in the mucus blanket as well as Clara cells in B-5 fixed materials, and only with the latter in formalin-fixed materials. Immuno-electron-microscopic study using protein・A-gold showed that the reaction products were accumulated in the electron dense granules of the Clara cells. These findings were considered to indicate that antigens we used were indeed produced in the Clara cells and were secreted into the airway. The antibodies were also reacted with those of the mice, but not with hamsters, guinea pigs, rabbits, dogs, cats, monkeys or the human. In this communication, our preparatory method of producing antibodies is described in detail
