中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2010年
3期
281-286
,共6页
韩凌斐%汪希鹏%王玲%熊诗诣%吕斯迹%艾贵海%洪岭%方勇%马丁
韓凌斐%汪希鵬%王玲%熊詩詣%呂斯跡%艾貴海%洪嶺%方勇%馬丁
한릉비%왕희붕%왕령%웅시예%려사적%애귀해%홍령%방용%마정
B/T淋巴细胞弱化因子%疱疹病毒进入介质%树突状细胞%共刺激分子
B/T淋巴細胞弱化因子%皰疹病毒進入介質%樹突狀細胞%共刺激分子
B/T림파세포약화인자%포진병독진입개질%수돌상세포%공자격분자
BTLA%HVEM%Dendritic cell%Costimulatory molecule
目的 探讨阻断BTLA-HVEM(B/T淋巴细胞弱化因子疱疹病毒进入介质)通路对树突状细胞功能的影响和相关免疫学机制.方法 构建小鼠BTLA胞外功能区的真核表达载体psBTLA,转染CHO细胞;HSP70-TC-1肿瘤抗原肽刺激小鼠骨髓来源DCs,流式细胞仪检测处理后DCs表面BTLA、HVEM的表达,同时给予转染了psBTLA质粒的CHO细胞的培养上清处理后,检测DCs表面B7-1的表达,ELISA检测上清中IL-12的分泌;处理后的DCs刺激脾细胞,检测淋巴细胞增殖和细胞因子分泌;检测psBTLA体内转染对宫颈癌细胞系TC-1成瘤小鼠DCs表达B7-1和肿瘤生长的影响.结果 成功构建小鼠BTLA胞外段的真核表达载体psBTLA,获得了稳定转染psBTLA的CHO细胞,在其培养上清检测到BTLA胞外段(sBTLA)的表达.DCs经抗原肽刺激后BTLA、HVEM表达均上调,加入含sBTLA的上清处理后上调B7-1,上清中分泌的IL-12增加,与脾细胞共培养时促进细胞增殖和IL-2、IFN-γ的分泌;体内基因转染psBTLA促进DCs表达B7-1以及抑制肿瘤生长.结论 通过sBTLA阻断BTLA-HVEM共抑制通路,可以进一步促进DCs的功能,更好地激活淋巴细胞,促进抗肿瘤免疫应答.
目的 探討阻斷BTLA-HVEM(B/T淋巴細胞弱化因子皰疹病毒進入介質)通路對樹突狀細胞功能的影響和相關免疫學機製.方法 構建小鼠BTLA胞外功能區的真覈錶達載體psBTLA,轉染CHO細胞;HSP70-TC-1腫瘤抗原肽刺激小鼠骨髓來源DCs,流式細胞儀檢測處理後DCs錶麵BTLA、HVEM的錶達,同時給予轉染瞭psBTLA質粒的CHO細胞的培養上清處理後,檢測DCs錶麵B7-1的錶達,ELISA檢測上清中IL-12的分泌;處理後的DCs刺激脾細胞,檢測淋巴細胞增殖和細胞因子分泌;檢測psBTLA體內轉染對宮頸癌細胞繫TC-1成瘤小鼠DCs錶達B7-1和腫瘤生長的影響.結果 成功構建小鼠BTLA胞外段的真覈錶達載體psBTLA,穫得瞭穩定轉染psBTLA的CHO細胞,在其培養上清檢測到BTLA胞外段(sBTLA)的錶達.DCs經抗原肽刺激後BTLA、HVEM錶達均上調,加入含sBTLA的上清處理後上調B7-1,上清中分泌的IL-12增加,與脾細胞共培養時促進細胞增殖和IL-2、IFN-γ的分泌;體內基因轉染psBTLA促進DCs錶達B7-1以及抑製腫瘤生長.結論 通過sBTLA阻斷BTLA-HVEM共抑製通路,可以進一步促進DCs的功能,更好地激活淋巴細胞,促進抗腫瘤免疫應答.
목적 탐토조단BTLA-HVEM(B/T림파세포약화인자포진병독진입개질)통로대수돌상세포공능적영향화상관면역학궤제.방법 구건소서BTLA포외공능구적진핵표체재체psBTLA,전염CHO세포;HSP70-TC-1종류항원태자격소서골수래원DCs,류식세포의검측처리후DCs표면BTLA、HVEM적표체,동시급여전염료psBTLA질립적CHO세포적배양상청처리후,검측DCs표면B7-1적표체,ELISA검측상청중IL-12적분비;처리후적DCs자격비세포,검측림파세포증식화세포인자분비;검측psBTLA체내전염대궁경암세포계TC-1성류소서DCs표체B7-1화종류생장적영향.결과 성공구건소서BTLA포외단적진핵표체재체psBTLA,획득료은정전염psBTLA적CHO세포,재기배양상청검측도BTLA포외단(sBTLA)적표체.DCs경항원태자격후BTLA、HVEM표체균상조,가입함sBTLA적상청처리후상조B7-1,상청중분비적IL-12증가,여비세포공배양시촉진세포증식화IL-2、IFN-γ적분비;체내기인전염psBTLA촉진DCs표체B7-1이급억제종류생장.결론 통과sBTLA조단BTLA-HVEM공억제통로,가이진일보촉진DCs적공능,경호지격활림파세포,촉진항종류면역응답.
Objective To explore the effect of blocking BTLA-HVEM (herpesvirus entry mediator-B and T lymphocyte attenuator) pathway on dendritic cell function and the related immunological mechanisms. Methods Murine BTLA extracellular domain eukaryotic expression vector psBTLA was constructed by gene recombination and transfected CHO by Lipofection method. Mouse bone marrow cells were induced to differentiate into DCs by GM-CSF plus IL-4. Expression of BTLA and HVEM on DCs was detected after HSPT0-TC-1 peptide complex stimulation by FACS. Expression of BT-1 and secretion of IL-12 were detected after HSP70-TC-1 peptide complex plus psBTLA transfected CHO culture supernatant stimulation on DCs. Pretreated DCs co-cultured with the same genetic background mouse splenocytes and lymphocytes proliferation and cytokine secretion were detected. Effect of psBTLA gene transfer in vivo on BT-1 expression of DCs and tumor growth on tumor-bearing mice was detected. Results Extracellular domain of murine BTLA was successfully constructed, psBTLA stable transfection CHO cells were obtained and expression of BTLA extracellular domain(sBTLA) was detected the in its culture supernatant. BTLA and HVEM expression of DCs were increased after stimulation by the antigen peptide complex. When DCs were treated with antigen peptide complex plus culture supernatant containing sBTLA, B7-1 expression and IL-12 secretion were increased. Co-cultured with splenocytes, lymphocytes proliferation and cytokine secretion, such as IL-2 and IFN-γ,, were also increased. Gene transfection with psBTLA in vivo promoted B7-1 expression on DCs and inhibited cervical cancer cells growth. Conclusion Blockade of BTLA-HVEM inhibitory pathway with sBTLA can further improve DCs function, activation of lymphocytes and promote antitumor immune response.
