Elson C O, Ealding W, Lefkowitz J
J Immunol Methods. 1984 Feb 24;67(1):101-8. doi: 10.1016/0022-1759(84)90089-9.
Mouse intestinal secretions can be readily obtained without harm to the mice by administering a lavage solution to them intragastrically followed by pilocarpine intraperitoneally. These secretions are rich in proteases but this enzyme activity can be blocked by addition of a mixture of inhibitors. Both total and specific IgA antibody could be measured in these secretions using ELISA techniques. The total IgA recovered was found to vary considerably, even in the same group of mice sampled on multiple occasions. Specific IgA anti-cholera toxin antibody was easily demonstrable in the intestinal secretions of mice fed cholera toxin but not of mice fed an irrelevant antigen. Expression of the specific IgA antibody per unit of total IgA recovered is desirable in order to correct for the variable recovery of IgA.
通过向小鼠胃内灌洗溶液,随后腹腔注射毛果芸香碱,可轻松获取小鼠肠道分泌物,且不会对小鼠造成伤害。这些分泌物富含蛋白酶,但通过添加抑制剂混合物可阻断这种酶活性。使用酶联免疫吸附测定(ELISA)技术可在这些分泌物中检测总IgA抗体和特异性IgA抗体。即使在多次采样的同一组小鼠中,发现回收的总IgA也有很大差异。在喂食霍乱毒素的小鼠肠道分泌物中可轻易检测到特异性抗霍乱毒素IgA抗体,而在喂食无关抗原的小鼠肠道分泌物中则检测不到。为校正IgA回收量的变化,每单位回收的总IgA中特异性IgA抗体的表达是可取的。