中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2008年
7期
425-429
,共5页
郭艳芳%马丽英%张跃新%袁林%孙坚萍%徐维四%赵全壁%屈水令%黄洋%邵一鸣
郭豔芳%馬麗英%張躍新%袁林%孫堅萍%徐維四%趙全壁%屈水令%黃洋%邵一鳴
곽염방%마려영%장약신%원림%손견평%서유사%조전벽%굴수령%황양%소일명
HIV-1%受体,细胞表面%受体,CCR5%受体,CXCR4
HIV-1%受體,細胞錶麵%受體,CCR5%受體,CXCR4
HIV-1%수체,세포표면%수체,CCR5%수체,CXCR4
HIV-1%Receptors,cell surface%Receptors,CCR5%Receptors,CXCR4
目的 研究HIV-1感染者不同时期分离的R5毒株的生物学特性.方法 采用传统的共培养方法分离并培养HIV-1,用表达CD4和CC趋化因子受体5(CCR5)或CXC趋化因子受体4(CXCR4)的GHOST细胞系,通过流式细胞仪测定病毒辅助受体的利用和感染性,从而判断所分离毒株的CCR5嗜型(R5型毒株);使用2 ng P24病毒量感染正常人分离的外周血单个核细胞(PBMC),ELISA法检测第1、3、5、7、10、15天的HIV-1 P24抗原,反映病毒复制能力;采用HIV-1核酸荧光定量检测试剂盒测定血浆病毒载量.数据分析采用t检验.结果 HIV-1B'亚型感染者22例,其中CD4+细胞>0.2×109/L和CD4+细胞≤0.2×109/L各11例;所分离的病毒仅利用CCR5辅助受体,均为R5型毒株,感染性的结果显示,来自CD4+细胞≤0.2×109/L的11株R5毒株的感染性为(7.392 7±4.584 2)%,而CD4+细胞>0.2×109/L的为(2.613 6±1.610 5)%,差异有统计学意义(t=3.262,P<0.05);两组病毒复制滴度在第7天开始明显上升,培养第7、10、15天,两组病毒复制动力学差异有统计学意义(t值分别为3.771、2.509和2.260;P<0.05),CD4+细胞≤0.2×109/L的R5毒株的复制能力较CD4+细胞>0.2×109/L的明显增强;CD4+细胞≤0.2 × 109/L R5型毒株的病毒载量的对数值为(5.606 8±0.815 1)拷贝/mL,CD4+细胞>0.2 × 109/L的为(4.729 8±0.431 6)拷贝/mL,两组差异有统计学意义(t=3.771,P<0.05).结论 疾病进展过程中,即使病毒的辅助受体利用未从CCR5转变为CXCR4,但病毒的感染性和复制能力已有明显改变.
目的 研究HIV-1感染者不同時期分離的R5毒株的生物學特性.方法 採用傳統的共培養方法分離併培養HIV-1,用錶達CD4和CC趨化因子受體5(CCR5)或CXC趨化因子受體4(CXCR4)的GHOST細胞繫,通過流式細胞儀測定病毒輔助受體的利用和感染性,從而判斷所分離毒株的CCR5嗜型(R5型毒株);使用2 ng P24病毒量感染正常人分離的外週血單箇覈細胞(PBMC),ELISA法檢測第1、3、5、7、10、15天的HIV-1 P24抗原,反映病毒複製能力;採用HIV-1覈痠熒光定量檢測試劑盒測定血漿病毒載量.數據分析採用t檢驗.結果 HIV-1B'亞型感染者22例,其中CD4+細胞>0.2×109/L和CD4+細胞≤0.2×109/L各11例;所分離的病毒僅利用CCR5輔助受體,均為R5型毒株,感染性的結果顯示,來自CD4+細胞≤0.2×109/L的11株R5毒株的感染性為(7.392 7±4.584 2)%,而CD4+細胞>0.2×109/L的為(2.613 6±1.610 5)%,差異有統計學意義(t=3.262,P<0.05);兩組病毒複製滴度在第7天開始明顯上升,培養第7、10、15天,兩組病毒複製動力學差異有統計學意義(t值分彆為3.771、2.509和2.260;P<0.05),CD4+細胞≤0.2×109/L的R5毒株的複製能力較CD4+細胞>0.2×109/L的明顯增彊;CD4+細胞≤0.2 × 109/L R5型毒株的病毒載量的對數值為(5.606 8±0.815 1)拷貝/mL,CD4+細胞>0.2 × 109/L的為(4.729 8±0.431 6)拷貝/mL,兩組差異有統計學意義(t=3.771,P<0.05).結論 疾病進展過程中,即使病毒的輔助受體利用未從CCR5轉變為CXCR4,但病毒的感染性和複製能力已有明顯改變.
목적 연구HIV-1감염자불동시기분리적R5독주적생물학특성.방법 채용전통적공배양방법분리병배양HIV-1,용표체CD4화CC추화인자수체5(CCR5)혹CXC추화인자수체4(CXCR4)적GHOST세포계,통과류식세포의측정병독보조수체적이용화감염성,종이판단소분리독주적CCR5기형(R5형독주);사용2 ng P24병독량감염정상인분리적외주혈단개핵세포(PBMC),ELISA법검측제1、3、5、7、10、15천적HIV-1 P24항원,반영병독복제능력;채용HIV-1핵산형광정량검측시제합측정혈장병독재량.수거분석채용t검험.결과 HIV-1B'아형감염자22례,기중CD4+세포>0.2×109/L화CD4+세포≤0.2×109/L각11례;소분리적병독부이용CCR5보조수체,균위R5형독주,감염성적결과현시,래자CD4+세포≤0.2×109/L적11주R5독주적감염성위(7.392 7±4.584 2)%,이CD4+세포>0.2×109/L적위(2.613 6±1.610 5)%,차이유통계학의의(t=3.262,P<0.05);량조병독복제적도재제7천개시명현상승,배양제7、10、15천,량조병독복제동역학차이유통계학의의(t치분별위3.771、2.509화2.260;P<0.05),CD4+세포≤0.2×109/L적R5독주적복제능력교CD4+세포>0.2×109/L적명현증강;CD4+세포≤0.2 × 109/L R5형독주적병독재량적대수치위(5.606 8±0.815 1)고패/mL,CD4+세포>0.2 × 109/L적위(4.729 8±0.431 6)고패/mL,량조차이유통계학의의(t=3.771,P<0.05).결론 질병진전과정중,즉사병독적보조수체이용미종CCR5전변위CXCR4,단병독적감염성화복제능력이유명현개변.
Objective To study biological characteristics of R5 tropic human immunodeficiency virus (HIV)-1 strains in different disease stage. Methods Primary clinical viruses were isolated from fresh peripheral blood mononuclear cells (PBMC) using co-culture methods; meanwhile, viral co receptor usage and infectivity were tested using flow cytometry on GHOST (3) cell lines,which expressed CD4 receptor and CC ehemokine receptor 5 (CCR5) or CXCR4 eoreceptor; to identified CCR5 tropic viruses(R5 tropic strains). Viral replication kinetics was detected in PBMCs. Plasma viral load was measured using an HIV-1 nucleotide fluorescence quantification assay kit. Results There were 22 individuals with HIV-1 subtype B' infection, in which 11 were CD4>0. 2 × 109/L and 11 were CD4≤0. 2 × 109/L. All isolated viruses used CCR5 coreceptor and therefore were HIV-1 R5 tropic strains. The infectivity of R5 tropic strains isolated from patients with CD4≤0.2 × 109/L was (7.392 7 ± 4. 584 2) % ; while the infectivity of R5 tropic strain from patients with CD4>0. 2 × 109/L was (2. 613 6 ± 1. 610 5)%. There were significant statistical difference(t= 3. 262, P<0.05). The possibility of viral replication became strong after the day 7 post-infection. There was a significant difference of viral replication between two groups in the day 7,10, 15 post-infection(t value was 3. 771, 2. 509 and 2. 260 respectively, P<0. 05). The possibility of viral replication was higher in CD4≤0.2 ×109/L group than that of CD4>0.2 × 109/L group. The logarithm of viral load was (5. 606 8 ± 0. 815 1 ) copies/mL in CD4≤0.2 × 109/L group and (4. 729 8 ± 0. 431 6) copies/mL in CD4> 0.2 × 109/L group. There was a significant difference between two groups(t = 3. 771 ; P<0.05). Conclusion Viral infection and replication are enhanced during progression of disease, even if viral coreceptor usage do not switch from CCR5 to CXCR4.