肾组织内淋巴细胞测定方法探讨

The Study of Detecting Technics for Lymphocyte Subsets in Renal Tissue

应用免疫组化法对肾组织中淋巴细胞亚群进行测定,以探讨肾组织标本处理、固定液选用、单克隆抗体作用时间对染色结果的影响。并比较了四种免疫组化方法的优劣。结果认为冰冻切片的肾组织经丙酮固定,加高稀释度CD_4、CD_(?)、B 细胞单克隆抗体作用1.5小时,再以标记辣根过氧化物酶的兔抗鼠、猪抗兔免疫球蛋白相继作用后,再加兔PAP,即改良PAP 四层法最适用于肾组织中淋巴细胞的测定。

Technics in using monoclonal antibodies to and identify assess the lympho-cyte subsets in renal tissus was investigated.A variety of processing for renal tissue,theselection of fixative solution and the a dequate time of monoclonal antibodies incubationin four different approaches of immunohistochemical technics were compared.It was foundthat 3-um frozen tissue sections fixed by acetone,then incubated with monoclonal antibo-dies(CD4,CD8 and B cell)for 1.5h,later the sections were sequentially incuqated withthe following antisera:peroxidase-conjugated rabbit antimouse immunoglobulins;peroxidase-conjugated swine antirabbit immunoglobulins and peroxidase-antiperoxidase complexesproduced in the rabbit(PAP).This modified 4-layer PAP method was superior to altern-ative method for lymphocyte subsets in renal tissue.