肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2009年
10期
654-656,659
,共4页
膀胱肿瘤%卡介苗%细胞毒性%免疫
膀胱腫瘤%卡介苗%細胞毒性%免疫
방광종류%잡개묘%세포독성%면역
Urinary bladder neoplasms%BCG vaccine%Cytotoxicity%immunologic
目的 探讨分泌人共刺激分子B7-2的重组卡介苗(rBCG)对人外周血单个核细胞(PBMC)IFN-α表达、免疫增强及杀伤膀胱癌细胞的作用.方法 SDS-PAGE和ELISA检测rBCG的表达产物hB7-2;rBCG和野生型BCG(wBCG)分别与PBMC共同培养,以单纯PBMC为对照,在不同时段收集培养液上清,经ELISA法检测上清液中IFN-α的表达水平;将BCG激活杀伤细胞(BCG activated killer cell,BAK细胞)与人膀胱癌EJ细胞共同培养,采用乳酸脱氧酶(LDH)释放试验检测活化免疫细胞对膀胱癌细胞的杀伤作用.结果 ELISA法检测出培养上清液hB7-2含量为3.8 U/ml.rBCG诱导PBMC产生细胞因子的表达明显高于同浓度的wBCG组(P<0.05);rBCG诱导的PBMC抗癌效应高于对照组诱导的抗癌效果(P<0.05).结论 rBCG诱导人PBMC高表达IFN-α,并通过增强人PBMC细胞的作用来提高抗膀胱肿瘤作用.
目的 探討分泌人共刺激分子B7-2的重組卡介苗(rBCG)對人外週血單箇覈細胞(PBMC)IFN-α錶達、免疫增彊及殺傷膀胱癌細胞的作用.方法 SDS-PAGE和ELISA檢測rBCG的錶達產物hB7-2;rBCG和野生型BCG(wBCG)分彆與PBMC共同培養,以單純PBMC為對照,在不同時段收集培養液上清,經ELISA法檢測上清液中IFN-α的錶達水平;將BCG激活殺傷細胞(BCG activated killer cell,BAK細胞)與人膀胱癌EJ細胞共同培養,採用乳痠脫氧酶(LDH)釋放試驗檢測活化免疫細胞對膀胱癌細胞的殺傷作用.結果 ELISA法檢測齣培養上清液hB7-2含量為3.8 U/ml.rBCG誘導PBMC產生細胞因子的錶達明顯高于同濃度的wBCG組(P<0.05);rBCG誘導的PBMC抗癌效應高于對照組誘導的抗癌效果(P<0.05).結論 rBCG誘導人PBMC高錶達IFN-α,併通過增彊人PBMC細胞的作用來提高抗膀胱腫瘤作用.
목적 탐토분비인공자격분자B7-2적중조잡개묘(rBCG)대인외주혈단개핵세포(PBMC)IFN-α표체、면역증강급살상방광암세포적작용.방법 SDS-PAGE화ELISA검측rBCG적표체산물hB7-2;rBCG화야생형BCG(wBCG)분별여PBMC공동배양,이단순PBMC위대조,재불동시단수집배양액상청,경ELISA법검측상청액중IFN-α적표체수평;장BCG격활살상세포(BCG activated killer cell,BAK세포)여인방광암EJ세포공동배양,채용유산탈양매(LDH)석방시험검측활화면역세포대방광암세포적살상작용.결과 ELISA법검측출배양상청액hB7-2함량위3.8 U/ml.rBCG유도PBMC산생세포인자적표체명현고우동농도적wBCG조(P<0.05);rBCG유도적PBMC항암효응고우대조조유도적항암효과(P<0.05).결론 rBCG유도인PBMC고표체IFN-α,병통과증강인PBMC세포적작용래제고항방광종류작용.
Objective To investigate the expression of IFN-α of the peripheral blood monocytes (PBMC) stimulated by bacille Calmeue-Guérin (BCG) expressing recombinant human B7-2, and the antitumor effect of BCG activated killer cells(BAK). Methods Expression of human B7-2 was detected by SDS-PAGE and ELISA. Recombinant BCG and wild-type BCG were used to stimulate PBMC in different concentrations in vitro. Supernatant was collected at various time points and IFN-α was detected by an enzyme-linked immunosorbent assay (ELISA). MTT assay was used to observe the effects of recombinant BCG on proliferation of T cell, and LDH assay was used to study antitumor cytotoxicity of BAK cells. Results The concentration of human B7-2 in culture was 3.8 U/ml by ELISA. Compared with wild-type BCG, recombinant BCG can induce more IFN-α. The results of the LDH release assay showed that the anti-tumor activity of BAK cells stimulated by recombinant BCG was 2.14 fold higher than that of wild-type BCG. Conclusion The expression of IFN-α in PBMC stimulated by recombinant BCG is higher than that stimulated by wild-type BCG, suggesting that enhanced antitumor activity of BAK when bladder cancer cells could be enchanced by using recombinant BCG.
