免疫血清对旋毛虫感染性幼虫侵入肠上皮细胞及其发育的影响  

In Vitro Effect of Immune Sera on the Invasion of Trichinella spiralis Infective Larvae into Intestinal Epithelial Cells and Their Development

作　　者：王书伟[1] 崔晶[1] 王中全[1] 王莉 

机构地区：[1]郑州大学医学院寄生虫学教研室,郑州450052

出　　处：《中国寄生虫学与寄生虫病杂志》2010年第5期348-352,共5页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No30972579);高校博士点专项科研基金(No20094101110004)~~

摘　　要：目的观察旋毛虫感染性幼虫排泄分泌(ES)抗原与旋毛虫感染性幼虫表面抗原免疫鼠血清对幼虫侵入HCT-8肠上皮细胞及其发育的影响。方法将旋毛虫感染性幼虫接种至半固体培养基(RPMI1640培养基+1.75%琼脂糖)+HCT-8细胞中,37℃5%CO2培养12、24、36、72和96h后镜下观察幼虫发育情况;将幼虫接种至含免疫血清的半固体培养基+HCT-8细胞中,培养15min后镜下观察幼虫形态及其对肠上皮细胞的侵入情况,36h后应用间接荧光抗体试验(IFAT)观察幼虫蜕皮,并计数Ⅰ期和Ⅱ～Ⅳ期幼虫。结果幼虫在半固体培养基培养12h可侵入HCT-8细胞单层,36～72h幼虫可蜕皮1～2次,培养96h可见早期成虫。在含ES抗原免疫血清与感染鼠血清条件下培养15min,幼虫头端可见免疫复合物形成的帽样结构,但在含表面抗原免疫血清、正常鼠血清或不含免疫血清条件下培养的幼虫头端则无帽样结构,头端带有帽样结构的幼虫不能侵入HCT-8细胞单层。在含ES抗原与表面抗原免疫血清条件下发育至Ⅱ～Ⅳ期幼虫的百分比(2.25%、2.2%)均明显低于含正常鼠血清条件下培养的幼虫(24.7%)(P<0.05)。结论旋毛虫ES抗原免疫血清可阻止幼虫对肠上皮细胞的侵入,ES抗原及表面抗原免疫血清均可阻止部分幼虫的发育(蜕皮)。Objective To observe the effect of sera from mice immunized with excretory-secretory （ES） antigen or surface antigen of Trichinella spiralis larvae on the invasion of intestinal epithelial cells in vitro by the larvae and on their development. Methods HCT-8 cells grown to confluence were overlaid with the larvae suspended in semisolid medium （RPMI 1640 medium ＋1.75% agarose）, and the larvae were then observed by using an inverted microscope after being incubated at 37 ℃ under 5% CO2 for 12、 24、 36、 72、 96 h. The larval development and its invasion into intestinal epithelial cells were observed under inverted microscope after 15 min when HCT-8 cells were overlaid with the larvae suspended in semisolid medium supplemented with immune sera. Finally, the 1st stage and 2nd～4th stage larvae were observed and enumerated by indirect fluorescent antibody test （IFAT） after 36 h incubation. Results When the larvae were cultured in semisolid medium, they invaded the HCT-8 cell monolayer and molted 1-2 times at 36 h to 72 h of culture. Early adult was observed at 96 h of culture. Cephalic caps on larvae were found at 15 min of culture when the larvae were cultured with medium containing immune sera, but the caps were not observed on those cultured with sera of normal mice or without sera. And the larvae with cephalic caps did not invade the cell monolayer. When the larvae were cultured with immune sera for 36 h, the percentage of 2nd～4th stage larvae （2.25%, 2.20%） were significantly lower than that of those cultured in normal sera （24.7%） （P0.05）. Conclusion The sera from mice immunized with excretorysecretory antigen or surface antigen of T. spiralis larvae prevent the invasion of the larvae into intestinal epithelial cells in vitro and impede the development （ecdysis） of the larvae.

关 键 词：旋毛虫 排泄分泌抗原 表面抗原 免疫血清 肠上皮细胞 

分 类 号：R383.15[医药卫生—医学寄生虫学]