中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2015年
1期
23-27
,共5页
彭松%贝俊杰%胡厚源%陈强
彭鬆%貝俊傑%鬍厚源%陳彊
팽송%패준걸%호후원%진강
蜱抗凝血肽%金黄色葡萄球菌超抗原样蛋白5%融合蛋白%血小板%淋巴细胞%P-选择素糖蛋白配体1
蜱抗凝血肽%金黃色葡萄毬菌超抗原樣蛋白5%融閤蛋白%血小闆%淋巴細胞%P-選擇素糖蛋白配體1
비항응혈태%금황색포도구균초항원양단백5%융합단백%혈소판%림파세포%P-선택소당단백배체1
KEY WORDS] Tick anticoagulant peptide%Staphylococcal superantigen like protein 5%Fusion protein%Platelets%Lymphocytes%P-selectin glycoprotein ligand 1
目的:研究抗炎、抗凝双效融合蛋白蜱抗凝血肽(TAP)-金黄色葡萄球菌超抗原样蛋白5(SSL5)对激活的血小板与人淋巴细胞结合作用的影响。方法:采用免疫磁珠分选法筛选人外周血总淋巴细胞;CCK-8法检测TAP-SSL5对细胞活力的影响;流式细胞术检测Jurkat细胞(人外周血白血病T细胞株)表面CD162( PSGL-1)的表达及TAP-SSL5对小鼠抗人CD162单抗( KPL-1)与Jurkat细胞结合的抑制作用。以20μmol/L ADP激活人血小板,流式细胞术检测血小板与Jurkat 细胞或人淋巴细胞的结合情况,并研究TAP-SSL5的干预作用。结果:30 mg/L及以下浓度的TAP-SSL5对Jurkat细胞的活力无明显影响。流式细胞术检测显示,10 mg/L的TAP-SSL5能显著抑制KPL-1与Jurkat细胞的结合;20μmol/L ADP 激活的血小板与Jurkat 细胞或淋巴细胞的结合率分别为(11.86±4.49)%和(8.32±1.00)%;细胞经10 mg/L TAP-SSL5预先处理后,结合率分别降至(6.73±2.71)%和(5.51±0.70)%,差异有统计学意义。结论:TAP-SSL5可与淋巴细胞表面的PSGL-1结合,从而抑制激活的血小板与人淋巴细胞的结合,这可能是抗炎、抗凝双效融合蛋白TAP-SSL5发挥其抗炎作用的机制之一。
目的:研究抗炎、抗凝雙效融閤蛋白蜱抗凝血肽(TAP)-金黃色葡萄毬菌超抗原樣蛋白5(SSL5)對激活的血小闆與人淋巴細胞結閤作用的影響。方法:採用免疫磁珠分選法篩選人外週血總淋巴細胞;CCK-8法檢測TAP-SSL5對細胞活力的影響;流式細胞術檢測Jurkat細胞(人外週血白血病T細胞株)錶麵CD162( PSGL-1)的錶達及TAP-SSL5對小鼠抗人CD162單抗( KPL-1)與Jurkat細胞結閤的抑製作用。以20μmol/L ADP激活人血小闆,流式細胞術檢測血小闆與Jurkat 細胞或人淋巴細胞的結閤情況,併研究TAP-SSL5的榦預作用。結果:30 mg/L及以下濃度的TAP-SSL5對Jurkat細胞的活力無明顯影響。流式細胞術檢測顯示,10 mg/L的TAP-SSL5能顯著抑製KPL-1與Jurkat細胞的結閤;20μmol/L ADP 激活的血小闆與Jurkat 細胞或淋巴細胞的結閤率分彆為(11.86±4.49)%和(8.32±1.00)%;細胞經10 mg/L TAP-SSL5預先處理後,結閤率分彆降至(6.73±2.71)%和(5.51±0.70)%,差異有統計學意義。結論:TAP-SSL5可與淋巴細胞錶麵的PSGL-1結閤,從而抑製激活的血小闆與人淋巴細胞的結閤,這可能是抗炎、抗凝雙效融閤蛋白TAP-SSL5髮揮其抗炎作用的機製之一。
목적:연구항염、항응쌍효융합단백비항응혈태(TAP)-금황색포도구균초항원양단백5(SSL5)대격활적혈소판여인림파세포결합작용적영향。방법:채용면역자주분선법사선인외주혈총림파세포;CCK-8법검측TAP-SSL5대세포활력적영향;류식세포술검측Jurkat세포(인외주혈백혈병T세포주)표면CD162( PSGL-1)적표체급TAP-SSL5대소서항인CD162단항( KPL-1)여Jurkat세포결합적억제작용。이20μmol/L ADP격활인혈소판,류식세포술검측혈소판여Jurkat 세포혹인림파세포적결합정황,병연구TAP-SSL5적간예작용。결과:30 mg/L급이하농도적TAP-SSL5대Jurkat세포적활력무명현영향。류식세포술검측현시,10 mg/L적TAP-SSL5능현저억제KPL-1여Jurkat세포적결합;20μmol/L ADP 격활적혈소판여Jurkat 세포혹림파세포적결합솔분별위(11.86±4.49)%화(8.32±1.00)%;세포경10 mg/L TAP-SSL5예선처리후,결합솔분별강지(6.73±2.71)%화(5.51±0.70)%,차이유통계학의의。결론:TAP-SSL5가여림파세포표면적PSGL-1결합,종이억제격활적혈소판여인림파세포적결합,저가능시항염、항응쌍효융합단백TAP-SSL5발휘기항염작용적궤제지일。
AIM: To study the effect of tick anticoagulant peptide-staphylococcal superantigen like protein 5 (TAP-SSL5), an anti-inflammatory and anticoagulant fusion protein , on the binding of activated platelets to human lym-phocytes.METHODS:Human periphery lymphocytes were isolated by magnetic activated cell sorting (MACS).The toxic-ity of TAP-SSL5 on the viability of Jurkat cell was assessed by CCK-8 assay.Flow cytometry was applied to detect the ex-pression of CD162 (PSGL-1) on the Jurkat cells (human peripheral blood leukemia T lymphocyte cell line ) and the inhibi-tory effect of TAP-SSL5 on the binding of mouse anti-human CD162 monoclonal antibody (KPL-1) to Jurkat cells.Platelets were activated by ADP at concentration of 20μmol/L, the binding rates of activated platelets to Jurkat cells or human lym-phocytes were assayed by flow cytometry .RESULTS:The concentration of TAP-SSL5 below 30 mg/L didn’ t affect the vi-ability of Jurkat cells .TAP-SSL5 at 10 mg/L competitively inhibited KPL-1 binding to Jurkat cells .The binding rates of activated platelets to Jurkat cells or lymphocytes were (11.86 ±4.49)% and (8.32 ±1.00)%, respectively, which de-creased to (6.73 ±2.71)%and (5.51 ±0.70)%after the Jurkat cells and lymphocytes were pre-incubated with 10 mg/L TAP-SSL5 (P <0.05).CONCLUSION:TAP-SSL5 binds to PSGL-1 expressed on lymphocyte surface and directly in-hibits the binding of activated platelets to human lymphocytes , which may be one of the anti-inflammatory mechanisms of TAP-SSL5.