中国人兽共患病学报
中國人獸共患病學報
중국인수공환병학보
CHINESE JOURNAL OF ZOONOSES
2014年
10期
1028-1032,1038
,共6页
日本血吸虫%重组Bb(pGEX-Sj26GST)疫苗%脾细胞增殖%T细胞亚群%脾细胞凋亡
日本血吸蟲%重組Bb(pGEX-Sj26GST)疫苗%脾細胞增殖%T細胞亞群%脾細胞凋亡
일본혈흡충%중조Bb(pGEX-Sj26GST)역묘%비세포증식%T세포아군%비세포조망
Schistosoma j aponicum%rBb ( pGEX-Sj26GST ) vaccine%splenocyte proliferation%T cell subsets%splenocyte apoptosis
目的：观察日本血吸虫重组Bb（pGEX-Sj26GST ）疫苗免疫BALB/c小鼠后脾细胞增殖、亚群和凋亡的动态变化。方法将疫苗分别经皮下注射（SC组）和鼻腔粘膜接种（IN组）免疫BALB/c鼠，在免疫后0～22周每2周每组随机剖杀4只小鼠，无菌取脾，制成单个悬浮脾细胞，用四甲基偶氮唑盐比色法（MTT法）检测脾细胞增殖水平、用流式细胞仪（FACsort）检测T细胞亚群和脾细胞凋亡状况。结果未刺激及SjAWA刺激时SC组小鼠脾细胞增值水平于免疫后4～20周明显升高， ConA刺激时于4～18周显著升高，均于免疫后8周达最高水平；IN组脾细胞增殖水平原液组、SjAWA组和ConA组分别于2～18周、2～10周及14～18周、2～8周和12～18周显著升高，均于免疫后4周达最大值（ P＜0．01或 P＜0．05）。SC组和IN组CD＋4 T细胞分别于免疫后2～14周、2周及6～16周升高，并于8周达峰值（ P＜0．01或 P＜0．05）；两组CD＋8 T 细胞均于2～20周轻微升高，分别于8周和6周达较高值（P＞0．05）。未刺激和ConA刺激时SC组脾细胞凋亡水平分别于免疫后2～4周、2～6周升高显著，均于免疫后2周达最大值（ P＜0．01或 P＜0．05）；IN组均于4周显著升高并达峰值（ P＜0．01）。结论日本血吸虫重组 Bb（pGEX-Sj26GST ）疫苗可诱导脾细胞的增殖，增加CD＋4 T细胞的数目，抑制脾细胞的凋亡而发挥保护性免疫应答。
目的：觀察日本血吸蟲重組Bb（pGEX-Sj26GST ）疫苗免疫BALB/c小鼠後脾細胞增殖、亞群和凋亡的動態變化。方法將疫苗分彆經皮下註射（SC組）和鼻腔粘膜接種（IN組）免疫BALB/c鼠，在免疫後0～22週每2週每組隨機剖殺4隻小鼠，無菌取脾，製成單箇懸浮脾細胞，用四甲基偶氮唑鹽比色法（MTT法）檢測脾細胞增殖水平、用流式細胞儀（FACsort）檢測T細胞亞群和脾細胞凋亡狀況。結果未刺激及SjAWA刺激時SC組小鼠脾細胞增值水平于免疫後4～20週明顯升高， ConA刺激時于4～18週顯著升高，均于免疫後8週達最高水平；IN組脾細胞增殖水平原液組、SjAWA組和ConA組分彆于2～18週、2～10週及14～18週、2～8週和12～18週顯著升高，均于免疫後4週達最大值（ P＜0．01或 P＜0．05）。SC組和IN組CD＋4 T細胞分彆于免疫後2～14週、2週及6～16週升高，併于8週達峰值（ P＜0．01或 P＜0．05）；兩組CD＋8 T 細胞均于2～20週輕微升高，分彆于8週和6週達較高值（P＞0．05）。未刺激和ConA刺激時SC組脾細胞凋亡水平分彆于免疫後2～4週、2～6週升高顯著，均于免疫後2週達最大值（ P＜0．01或 P＜0．05）；IN組均于4週顯著升高併達峰值（ P＜0．01）。結論日本血吸蟲重組 Bb（pGEX-Sj26GST ）疫苗可誘導脾細胞的增殖，增加CD＋4 T細胞的數目，抑製脾細胞的凋亡而髮揮保護性免疫應答。
목적：관찰일본혈흡충중조Bb（pGEX-Sj26GST ）역묘면역BALB/c소서후비세포증식、아군화조망적동태변화。방법장역묘분별경피하주사（SC조）화비강점막접충（IN조）면역BALB/c서，재면역후0～22주매2주매조수궤부살4지소서，무균취비，제성단개현부비세포，용사갑기우담서염비색법（MTT법）검측비세포증식수평、용류식세포의（FACsort）검측T세포아군화비세포조망상황。결과미자격급SjAWA자격시SC조소서비세포증치수평우면역후4～20주명현승고， ConA자격시우4～18주현저승고，균우면역후8주체최고수평；IN조비세포증식수평원액조、SjAWA조화ConA조분별우2～18주、2～10주급14～18주、2～8주화12～18주현저승고，균우면역후4주체최대치（ P＜0．01혹 P＜0．05）。SC조화IN조CD＋4 T세포분별우면역후2～14주、2주급6～16주승고，병우8주체봉치（ P＜0．01혹 P＜0．05）；량조CD＋8 T 세포균우2～20주경미승고，분별우8주화6주체교고치（P＞0．05）。미자격화ConA자격시SC조비세포조망수평분별우면역후2～4주、2～6주승고현저，균우면역후2주체최대치（ P＜0．01혹 P＜0．05）；IN조균우4주현저승고병체봉치（ P＜0．01）。결론일본혈흡충중조 Bb（pGEX-Sj26GST ）역묘가유도비세포적증식，증가CD＋4 T세포적수목，억제비세포적조망이발휘보호성면역응답。
To observe the dynamic changes of splenocyte proliferation ,subsets and apoptosis in mice immunized with re-combinantBifidobacteriumbifidum(pGEX-Sj26GST)ofSchistosomajaponicum,themiceweresubcutaneously(SCgroup) and intranasally (IN group) immunized ,respectively .Four mice from each group were sacrificed in every 2 wk during 0-20 wk after immunization .Splenocyte proliferation was investigated by MTT colorimetric assay ,subsets of CD+4 and CD+8 T cells and apoptosis of splenocytes by FACsort flow cytometry .In SC group ,unstimulated and stimulated with S jAWA ,the level of splenocyte proliferation significantly increased at 4-20 wk after vaccination and increased markedly at 4-18 wk stimulated with ConA ,both of which peaked at 8 wk;in IN group ,the proliferation level of splenocyte cultured with SjAWA and ConA signif-icantly increased during the 4-18 wk ,2-10 wk and 14-18 wk ,2-8 wk and 12-18 wk ,respectively ,and all reached the maximum at the 4 wk after immunization (P<0 .01 or P<0 .05) .CD+4 subsets increased obviously during 2-14 wk ,2 wk and 6-16 wk re-spectively ,and reached the peak at 8 wk (P<0 .01 or P<0 .05) in both group ,while CD8+ subsets rose lightly during 2-20 wk in both group ,and reached the maximum at 8 wk (SC group) and 6 wk respectively (P>0 .05) .Whether unstimulated or stimulated with ConA ,the level of splenocyte apoptosis of which remarkably increased at 2-4 wk and 2-6 wk separately in SC group ,and both peaked at 2 wk (P<0 .01 or P<0 .05 );in IN group ,the level of splenocyte apoptosis all increased at 4 wk and reached the maximum at the same time .In summary ,by inducing the proliferation of splenocytes ,increasing CD4 + T cells and decreasing splenocyte apoptosis ,the rBb (pGEX-Sj26GST ) vaccine plays a critical role in the protective immune response .
