乳牛L-选择素在中性粒细胞中表达与分布的定位研究

采用间接荧光标记及胶体金标记方法通过流式细胞仪和低压扫描电子显微镜,检测了L-选择素在健康泌乳中期乳牛(A组)及患金黄色葡萄球菌型临床型(B组)和亚临床型乳腺炎乳牛(C组)中性粒细胞上表达的基本规律。结果表明:三者表达L-选择素的中性粒细胞阳性率分别为97.90%、97.69%和96.67%(收集5 000个细胞),组间差异不显著(P>0.05),但患亚临床型乳腺炎乳牛的阳性率从3次测定的结果看是逐渐增加的;L-选择素表达的间接荧光强度组间差异极显著(P<0.01);而患亚临床型乳腺炎的乳牛组内差异也极显著(P<0.01);扫描电镜观察到的中性粒细胞表面脊样结构的分布及形态与流式细胞仪检测的结果具有相似性。

Localization of expression and distribution of L-selectin in neutrophil of cows

Basic regularity of L-selectin in PMN of healthy lactating cows(Group A),cows with clinical staphylococcus aureus mastitis(Group B) and subclinical staphylococcus aureus mastitis(Group C) was studied respectively by means of the indirect fluorescence labelling and the colloidal gold labelling with help of the scanning electron microscope and the flow cytometer.Positive rates of the PMN in which L-(selectin) gene was expressed in 3 groups were 97.90%,97.69% and 96.67%,respectively,and were not significantly different among the 3 groups.But the positive rates of the PMN of the cows with subclinical mastitis increased gradually.The indirect fluorescence intensity difference among the 3 groups was significant.The fluorescence intensity of the healthy lactating cows was not different by three tests,neither was that of the cows with clinical mastitis.The difference of fluorescence intensity of L-selectin among the 3 groups was significant.However,the fluorescence intensity of L-selectin was significantly different among the 3 cows suffering from subclinical mastitis.Scanning electron microscopy showed that the distribution and form of ridge structure on the surface of the PMN was similar to the results detected by the flow(cytometry.)