中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2010年
4期
267-271
,共5页
杨德红%刘文佳%陈敏%吴育美%邹晓平
楊德紅%劉文佳%陳敏%吳育美%鄒曉平
양덕홍%류문가%진민%오육미%추효평
胰腺肿瘤%树突细胞%卡介苗
胰腺腫瘤%樹突細胞%卡介苗
이선종류%수돌세포%잡개묘
Pancreatic neoplasms%Dendritic cells%CGB vaccine
目的 探讨负载胰腺癌细胞株PANC1裂解产物(tumor lysate,TL)的树突状细胞(dendritic cells,DC)经卡介苗(CGB)活化后诱导的自体淋巴细胞体外抗胰腺癌效应.方法 应用rhGM-CSF和rhIL-4从健康人外周血单个核细胞诱导培养DC,用TL负载DC,并用CGB促其成熟.倒置相差显微镜观察DC形态,流式细胞仪检测细胞CD1a、CD83、CD86及HLA-DR表型,ELISA法检测培养上清液中TNF-α和IL-12p70含量.CCK-8检测DC诱导自体混合淋巴细胞的增殖率以及活化淋巴细胞对PANCl、PaTu8988及SCG7901细胞的杀伤率.结果 CGB活化负载TL的DC后,CD83和CD86阳性率分别从(3.7±0.3)%和(38.6±5.0)%显著增加至(16.5±0.6)%和(76.5±2.5)%(P＜0.05);DC分泌的IL-12p70和TNF-α量分别从(20.18±2.06)pg/ml和(61.38±1.19)pg/ml显著增加至(62.48±3.80)pg/ml和(592.53±17.96)pg/ml(P＜0.01);DC与自体混合淋巴细胞以1∶100、1∶50、1∶10、1∶5的比例共培养,淋巴细胞增殖率分别从(3.90±1.41)%、(4.07±0.40)%、(3.39±1.05)%、(2.82±0.39)%显著增加至(55.38±3.58)%、(75.00±2.54)%、(77.07±3.40)%、(99.07±2.40)%(P＜0.01);DC活化淋巴细胞对PANC1细胞的杀伤率在效靶比为1∶5、1∶10、1∶20、1∶50时分别可达(71.73±0.46)%、(49.44±0.98)%、(31.76±2.77)%、(19.03±3.04)%,但对PaTu8988和SCG7901细胞的杀伤率显著降低.结论经CGB活化的胰腺癌DC疫苗成熟度增加,体外表现出高效特异的抗胰腺癌细胞的效应.
目的 探討負載胰腺癌細胞株PANC1裂解產物(tumor lysate,TL)的樹突狀細胞(dendritic cells,DC)經卡介苗(CGB)活化後誘導的自體淋巴細胞體外抗胰腺癌效應.方法 應用rhGM-CSF和rhIL-4從健康人外週血單箇覈細胞誘導培養DC,用TL負載DC,併用CGB促其成熟.倒置相差顯微鏡觀察DC形態,流式細胞儀檢測細胞CD1a、CD83、CD86及HLA-DR錶型,ELISA法檢測培養上清液中TNF-α和IL-12p70含量.CCK-8檢測DC誘導自體混閤淋巴細胞的增殖率以及活化淋巴細胞對PANCl、PaTu8988及SCG7901細胞的殺傷率.結果 CGB活化負載TL的DC後,CD83和CD86暘性率分彆從(3.7±0.3)%和(38.6±5.0)%顯著增加至(16.5±0.6)%和(76.5±2.5)%(P＜0.05);DC分泌的IL-12p70和TNF-α量分彆從(20.18±2.06)pg/ml和(61.38±1.19)pg/ml顯著增加至(62.48±3.80)pg/ml和(592.53±17.96)pg/ml(P＜0.01);DC與自體混閤淋巴細胞以1∶100、1∶50、1∶10、1∶5的比例共培養,淋巴細胞增殖率分彆從(3.90±1.41)%、(4.07±0.40)%、(3.39±1.05)%、(2.82±0.39)%顯著增加至(55.38±3.58)%、(75.00±2.54)%、(77.07±3.40)%、(99.07±2.40)%(P＜0.01);DC活化淋巴細胞對PANC1細胞的殺傷率在效靶比為1∶5、1∶10、1∶20、1∶50時分彆可達(71.73±0.46)%、(49.44±0.98)%、(31.76±2.77)%、(19.03±3.04)%,但對PaTu8988和SCG7901細胞的殺傷率顯著降低.結論經CGB活化的胰腺癌DC疫苗成熟度增加,體外錶現齣高效特異的抗胰腺癌細胞的效應.
목적 탐토부재이선암세포주PANC1렬해산물(tumor lysate,TL)적수돌상세포(dendritic cells,DC)경잡개묘(CGB)활화후유도적자체림파세포체외항이선암효응.방법 응용rhGM-CSF화rhIL-4종건강인외주혈단개핵세포유도배양DC,용TL부재DC,병용CGB촉기성숙.도치상차현미경관찰DC형태,류식세포의검측세포CD1a、CD83、CD86급HLA-DR표형,ELISA법검측배양상청액중TNF-α화IL-12p70함량.CCK-8검측DC유도자체혼합림파세포적증식솔이급활화림파세포대PANCl、PaTu8988급SCG7901세포적살상솔.결과 CGB활화부재TL적DC후,CD83화CD86양성솔분별종(3.7±0.3)%화(38.6±5.0)%현저증가지(16.5±0.6)%화(76.5±2.5)%(P＜0.05);DC분비적IL-12p70화TNF-α량분별종(20.18±2.06)pg/ml화(61.38±1.19)pg/ml현저증가지(62.48±3.80)pg/ml화(592.53±17.96)pg/ml(P＜0.01);DC여자체혼합림파세포이1∶100、1∶50、1∶10、1∶5적비례공배양,림파세포증식솔분별종(3.90±1.41)%、(4.07±0.40)%、(3.39±1.05)%、(2.82±0.39)%현저증가지(55.38±3.58)%、(75.00±2.54)%、(77.07±3.40)%、(99.07±2.40)%(P＜0.01);DC활화림파세포대PANC1세포적살상솔재효파비위1∶5、1∶10、1∶20、1∶50시분별가체(71.73±0.46)%、(49.44±0.98)%、(31.76±2.77)%、(19.03±3.04)%,단대PaTu8988화SCG7901세포적살상솔현저강저.결론경CGB활화적이선암DC역묘성숙도증가,체외표현출고효특이적항이선암세포적효응.
Objective To evaluate in vitro anti tumor effect of host lymphocyte primed by CalmetteGuerin bacillus (CGB) activated dendritic cells (DC) based PANC1 lysate. Methods DCs were obtained from peripheral blood mononuclear cells of healthy volunteer and cuitured by rhGM CSF and rhIL 4. DC vaccines for pancreatic cancer were loaded with PANC1 tumor lysate (TL) and were further maturated by CGB.CD1a, CD83, CD86 and HLA-DR phenotype was characterized by flow cytometer, and IL-12p70 and TNF-α concentration in DC culture supernatant were measured by ELISA. Autologous mixed lymphocyte proliferation and the cytotoxicity of cytotoxic T lymphocytes primed by activated DCs to PANC1, PaTu8988 and SCG7901 tumor cells was measured by CCK 8 test. Results When DCs based PANC1 lysate were activated by CGB,the expression rates of CD83 and CD86 were increased from (3.7±0.3)% and (38.6±5.0)% to (16.5±0.6)% and (76.6±2.5)% (P ＜0.05 ). The concentrations of cytokines ILpl2p70 and TNF-α were increased from (20.18±2.06 ) pg/mland (61.38±1.19) pg/mlto (62.48±3.80) pg/mland (592.53±17.96)pg/ml (P＜0. 01 ). When co-cultured with CGB activated DCs based PANC1 lysate in proportion of 0.40)% , (3.39±1.05)% , (2.82±0.39)% significantly increased to (55.38±3.58)% , (75.0±2.54) % , (77.07±3.4)% , (99.07±2.4)% (P＜0.01) , respectively. The killing effects of lymphocytes 2.77)%, (19.03±3.04) %; but the killing effects on PaTu8988 and SCG7901 were significantly decreased.Conclusions DC vaccines for pancreatic cancer could be more maturated when activated by CGB, and could show a high capability of anti-tumor in vitro.
