间接免疫过氧化物酶技术鉴定猪和牛的肥大细胞  

IDENTIFICATION OF PORCINE AND BOVINE MAST CELLS BY AN INDIRECT IMMUNOPEROXIDASE TECHNIQUE *

作　　者：许乐仁[1,2,3,4] 江萍[1,2,3,4] M.M.卡尔 K.P.帕尔赛 A.W.瓦尔斯 G.A.霍尔 

机构地区：[1]贵州农学院动物科学系 [2]AFRC动物保健研究院康普顿实验室 [3]南开普敦大学免疫药理学组 [4]南开普敦总医院

出　　处：《动物学报》1997年第3期294-302,共9页ACTA ZOOLOGICA SINICA

摘　　要：用小鼠抗人肥大细胞类胰蛋白酶单克隆抗体ＡＡ１，ＡＡ３及ＡＡ５的间接免疫过氧化物酶技术对经Ｃａｒｎｏｙ液或中性缓冲福尔马林（ＮＢＦ）固定的猪和犊牛空肠、舌及胸腺的石蜡切片进行了免疫染色。对猪和牛的肥大细胞特异性免疫染色与常规的组织化学染色的结果进行了比较。ＮＢＦ或Ｃａｒｎｏｙ液固定和猪组织经单克隆抗体ＡＡ１，ＡＡ３及ＡＡ５免疫染色显现的肥大细胞在分布、形态和密度方面与甲苯胺蓝或阿尔辛蓝染色切片上的肥大细胞相似。猪组织采用Ｃａｒｎｏｙ液固定，ＡＡ５免疫染色效果最好；而采用ＮＢＦ固定，这三种抗体均可获得十分满意的效果。Ｃａｒｎｏｙ液固定的犊牛组织，ＡＡ３和ＡＡ５对所有的肥大细胞群体均可得到特异性强的染色，但这些抗体对ＮＢＦ固定的犊牛组织的免疫染色均不理想。小鼠抗人类肥大细胞类胰蛋白酶单克隆抗体与猪和牛的肥大细胞的交叉反应证实了这些动物的肥大细胞中均含有类胰蛋白酶故使用这些单克隆抗体的免疫酶技术可作为鉴定猪和牛肥大细胞的特异性方法。Murine monoclonal antibodies (mAbs) (AA1, AA3 and AA5), raised against human mast cell tryptase, were used in an indirect immunoperoxidase method to immunostain the paraffin sections of jejunum, tongue and thymus, collected from pigs and calves, fixed in Carnoy's fluid or neutral buffered formalin (NBF). The specific staining of porcine and bovine mast cells was compared with routine histochemical stainings. The distribution, morphology and density of mast cells in pig tissue fixed in NBF or Carnoy's fluid and immunostained with the mAbs AA1, AA3 and AA5 were similar to those seen when sections were stained with toluidine blue or Alcian blue. The mAb AA5 gave best staining of mast cells in pig tissues that had been fixed in Carnoy's fluid; CTMC were stained poorly by AA1 and AA3 and CTMC in piglets were not stained by any mAb. In calf tissue fixed in Carnoy's fluid, AA3 and AA5 gave specific and intense staining of all mast cell populations, but fixation in NBF was unsatisfactory for immunostaining with the mAbs. The cross-reaction of murine mAbs against human mast cell tryptase with porcine and bovine mast cells demonstrated that these mast cells contain tryptase and that the indirect immunoperoxidase technique using mAbs against human mast cell tryptase can be considered a specific method for the detection of porcine and bovine mast cells.

关 键 词：家畜免疫学 猪 牛肥大细胞 免疫染色 

分 类 号：S852.4[农业科学—基础兽医学]