慢性乙型肝炎病毒感染患者外周血特异性CD8~+T淋巴细胞检测的临床意义

目的研究慢性乙型肝炎病毒(HBV)感染患者外周血特异性CD8+T淋巴细胞(CTL)频率的变化及临床价值。方法采用乙型肝炎病毒核心抗原(HBcAg)18-27表位肽-人类白细胞抗原(HLA)-A*0201五聚体及CD8单克隆抗体,设计流式细胞技术检测HLA-A2+6例、HLA-A2-12例非HBV感染者、HLA-A2+49例和HLAA2-42例HBV感染者外周血中针对该肽段的特异性CTL数量,占总计数CD8+细胞数百分比表示。结果 HLA-A2-HBV感染未抗病毒治疗者特异性CTL(0.30%～14.40%,中位数1.13%,n=20)与HLA-A2-非HBV感染者(0.33%～3.90%,中位数1.04%,n=12),差异无统计学意义(P>0.05),但显著低于HLA-A2-抗病毒治疗者(0.25%～20.30%,中位数2.11%,n=22,P<0.05);HLA-A2-抗病毒治疗者特异性CTL和HLA-A2+未抗病毒治疗者特异性CTL(0.20%～29.90%,中位数2.22%,n=20)均显著高于HLA-A2-非HBV感染者(P<0.01);HLA-A2+抗病毒治疗者特异性CTL(0.14%～39.22%,中位数1.33%,n=29)显著高于HLA-A2-非HBV感染者(P<0.05)。HLA-A2+未抗病毒组中血清HBV DNA<103 copy/mL、丙氨酸氨基转移酶小于40U/L者特异性CTL频率增高(P<0.05、0.01)。结论 HBV抗原肽-HLA-A*0201五聚体流式细胞技术能在体外直接检测外周血HBV特异性CTL频率的变化,其水平一定程度反映不同临床感染状态慢性HBV感染患者T淋巴细胞对特异性抗原表位免疫应答的差异。     

载脂蛋白M单克隆抗体的制备及其初步应用

Objective To prepare anti-apoM monoantibodies with high affinity and high purity, and investigate apoM distribution among human tissues and different groups of people. Methods BALB/c mice were injected intracutaneously with recombinant apoM. After cytomixis, screening and cloning, we established a hybridoma, which grew well and steadily secreted antibodies. The ascites were acquired by injecting BALB/c mice intraperitoneally and anti-apoM monoantibodies were gained using standard techniques. We detected apoM levels in healthy individuals and the patients with coronary heart disease including stable angina (SA) group and acute coronary syndrome (ACS) group using the anti-apoM monoantibodies. ResultsThree anti-apoM monoantibodies were collected and confirmed after subtype identification and block test. The apoM protein were detected in some human cells and human tissues by these three monoantibodies. The concentration of apoM was (11.02 ±1.96) ×10 -3 g/L, (10. 76± 1.32) ×10-3 g/L, (12. 83 ± 2. 28) × 10-3 g/L in SA, ACS and control group respectively. There was significant difference within the three groups (F = 11. 544, P ＜ 0. 05). Comparing apoM concentrations among control group and coronary heart disease groups, it showed that the levels of apoM were lower in coronary heart disease groups than in control group(t =2. 962 and 3. 967,P ＜0. 05). There was no significant difference between two coronary heart disease groups (t = 1. 033, P ＞ 0. 05). Conclusion Anti-apoM monoantibodies are successfully raised and could combine with apoM in human cells and tissues. This lays the foundation for the apoM study in apolipoprotein metabolism.     

载脂蛋白M单克隆抗体的制备及其初步应用

Objective To prepare anti-apoM monoantibodies with high affinity and high purity, and investigate apoM distribution among human tissues and different groups of people. Methods BALB/c mice were injected intracutaneously with recombinant apoM. After cytomixis, screening and cloning, we established a hybridoma, which grew well and steadily secreted antibodies. The ascites were acquired by injecting BALB/c mice intraperitoneally and anti-apoM monoantibodies were gained using standard techniques. We detected apoM levels in healthy individuals and the patients with coronary heart disease including stable angina (SA) group and acute coronary syndrome (ACS) group using the anti-apoM monoantibodies. ResultsThree anti-apoM monoantibodies were collected and confirmed after subtype identification and block test. The apoM protein were detected in some human cells and human tissues by these three monoantibodies. The concentration of apoM was (11.02 ±1.96) ×10 -3 g/L, (10. 76± 1.32) ×10-3 g/L, (12. 83 ± 2. 28) × 10-3 g/L in SA, ACS and control group respectively. There was significant difference within the three groups (F = 11. 544, P ＜ 0. 05). Comparing apoM concentrations among control group and coronary heart disease groups, it showed that the levels of apoM were lower in coronary heart disease groups than in control group(t =2. 962 and 3. 967,P ＜0. 05). There was no significant difference between two coronary heart disease groups (t = 1. 033, P ＞ 0. 05). Conclusion Anti-apoM monoantibodies are successfully raised and could combine with apoM in human cells and tissues. This lays the foundation for the apoM study in apolipoprotein metabolism.     

测定血清肌红蛋白水平对西宁地区脓毒症患者病情及预后判断的应用价值

目的 探讨血清肌红蛋白(MYO)对西宁地区脓毒症患者病情及预后判断的应用价值.方法 选择青海大学附属医院急诊重症监护病房(EICU)30例脓毒症患者,24 h内测定血清MYO水平并进行急性生理学与慢性健康状况评分系统Ⅱ(APACHEⅡ)评分.采用集束化治疗策略,判定28 d患者预后.比较存活组与死亡组血中MYO和APACHEⅡ评分,并对MYO和APACHEⅡ评分进行相关性分析;按MYO将患者分为＜500(10例)、500～1 000(14例)、＞1 000 μg/L(6例)3组,比较各组APACHEⅡ评分和死亡例数.结果 30例患者中存活16例,死亡14例.存活组血清MYO和APACHEⅡ评分明显低于死亡组[MYO(μg/L):607.85±499.40比976.21±370.10;APACHEⅡ评分(分):15.50±4.43比18.93±3.63,t1=2.28,t2=2.29,均P<0.05].随着MYO升高,患者死亡例数明显增加(MYO＜500、500～1 000、＞1 000 μg/L组分别为2、7、5例,χ2=5.94,P<0.05),但APACHEⅡ评分差异无统计学意义.相关分析显示,血清MYO与APACHEⅡ评分呈明显正相关(r＝0.407,P<0.05).结论 测定MYO可反映西宁地区脓毒症患者的病情及预后.

Abstract：

Objective To investigate the value of determination of serum myoglobin (MYO) in estimation of the degree of illness and prognosis of patients with sepsis in Xining area. Methods Serum MYO was measured and acute physiological and chronic health estimationⅡ (APACHEⅡ) score was evaluated in 30 cases with sepsis within 24 hours of admission to emergency intensive care unit (EICU), and their correlation was analyzed. The patients were divided into two groups, survival group and death group according to the result within 28 days. The MYO and APACHEⅡ score were analyzed in both groups. All cases were divided into three groups: namely <500 (n=10), 5001 000 (n=14), >1 000 μg/L (n=6) groups, according to serum MYO value, and APACHEⅡ score and dead case were compared among three groups. Results Sixteen patients survived, and 14 patients died. The level of serum MYO and APACHEⅡ score were significantly lower in survival group than death group [MYO (μg/L): 607.85±499.40 vs. 976.21±370.10, APACHEⅡ score: 15.50±4.43 vs. 18.93±3.63, t1=2.28, t2=2.29, both P<0.05]. With the elevation of serum MYO, the dead case was increased in sepsis patients (the dead case in MYO<500, 5001 000, >1 000 μg/L groups was 2, 7 , 5 cases, respectively, χ2=5.94, P<0.05), but there was no difference in APACHEⅡ score among three groups. There was significal positive correlation between serum MYO and APACHEⅡ score (r＝0.407, P<0.05). Conclusion Determination of serum MYO can reflect degree of illness and prognosis of sepsis patients in Xining area.     

肺表面活性物质替代治疗与支气管肺发育不良的关系

Objective To explore, the effect of different dosage of pulmonary surfactant (PS) on the inci-dence of bronchopulmanary dysplasia. Method Four hundred and three premature infants with hyaline membrane disease were divided into 3 groups according to the dose of PS: low-dose group (L-PS group, ≤ 100 mg/kg, n =188) ,high-dose group(H-PS group, > 100 mg/kg, n = 94) and no-PS group (N-PS group, n = 121). The frac-tional inspired oxygen(FiO2) and ptlmonary oxygenating function before and after 6 hours treatment were observed and the durations of oxygen therapy and mechanical ventilation, frequency of repeated intubafion, length of hospi-talization and the incidence of BPD were compared among the three groups. Results After 6 hours PS administra-tion, the FiO2,oxygen index and duration of oxygen therapy and mechanical ventilation were significantly decreased (P <0.05), while PO2 and the arterio-alveolar partial pressure of oxygen were significantly increased (P <0.05)in the H-PS and L-PS groups, compared with the N-PS group. Compared with the L-PS and N-PS groups,the H-PS group showed a decreased incidence of BPD. Conclusions PS administration could improve the pul-monary oxygenation and prevent the development of BPD, especially in high-dose.     

肺表面活性物质替代治疗与支气管肺发育不良的关系

Objective To explore, the effect of different dosage of pulmonary surfactant (PS) on the inci-dence of bronchopulmanary dysplasia. Method Four hundred and three premature infants with hyaline membrane disease were divided into 3 groups according to the dose of PS: low-dose group (L-PS group, ≤ 100 mg/kg, n =188) ,high-dose group(H-PS group, > 100 mg/kg, n = 94) and no-PS group (N-PS group, n = 121). The frac-tional inspired oxygen(FiO2) and ptlmonary oxygenating function before and after 6 hours treatment were observed and the durations of oxygen therapy and mechanical ventilation, frequency of repeated intubafion, length of hospi-talization and the incidence of BPD were compared among the three groups. Results After 6 hours PS administra-tion, the FiO2,oxygen index and duration of oxygen therapy and mechanical ventilation were significantly decreased (P <0.05), while PO2 and the arterio-alveolar partial pressure of oxygen were significantly increased (P <0.05)in the H-PS and L-PS groups, compared with the N-PS group. Compared with the L-PS and N-PS groups,the H-PS group showed a decreased incidence of BPD. Conclusions PS administration could improve the pul-monary oxygenation and prevent the development of BPD, especially in high-dose.     

肺表面活性物质替代治疗与支气管肺发育不良的关系

Objective To explore, the effect of different dosage of pulmonary surfactant (PS) on the inci-dence of bronchopulmanary dysplasia. Method Four hundred and three premature infants with hyaline membrane disease were divided into 3 groups according to the dose of PS: low-dose group (L-PS group, ≤ 100 mg/kg, n =188) ,high-dose group(H-PS group, > 100 mg/kg, n = 94) and no-PS group (N-PS group, n = 121). The frac-tional inspired oxygen(FiO2) and ptlmonary oxygenating function before and after 6 hours treatment were observed and the durations of oxygen therapy and mechanical ventilation, frequency of repeated intubafion, length of hospi-talization and the incidence of BPD were compared among the three groups. Results After 6 hours PS administra-tion, the FiO2,oxygen index and duration of oxygen therapy and mechanical ventilation were significantly decreased (P <0.05), while PO2 and the arterio-alveolar partial pressure of oxygen were significantly increased (P <0.05)in the H-PS and L-PS groups, compared with the N-PS group. Compared with the L-PS and N-PS groups,the H-PS group showed a decreased incidence of BPD. Conclusions PS administration could improve the pul-monary oxygenation and prevent the development of BPD, especially in high-dose.     

肺表面活性物质替代治疗与支气管肺发育不良的关系

Objective To explore, the effect of different dosage of pulmonary surfactant (PS) on the inci-dence of bronchopulmanary dysplasia. Method Four hundred and three premature infants with hyaline membrane disease were divided into 3 groups according to the dose of PS: low-dose group (L-PS group, ≤ 100 mg/kg, n =188) ,high-dose group(H-PS group, > 100 mg/kg, n = 94) and no-PS group (N-PS group, n = 121). The frac-tional inspired oxygen(FiO2) and ptlmonary oxygenating function before and after 6 hours treatment were observed and the durations of oxygen therapy and mechanical ventilation, frequency of repeated intubafion, length of hospi-talization and the incidence of BPD were compared among the three groups. Results After 6 hours PS administra-tion, the FiO2,oxygen index and duration of oxygen therapy and mechanical ventilation were significantly decreased (P <0.05), while PO2 and the arterio-alveolar partial pressure of oxygen were significantly increased (P <0.05)in the H-PS and L-PS groups, compared with the N-PS group. Compared with the L-PS and N-PS groups,the H-PS group showed a decreased incidence of BPD. Conclusions PS administration could improve the pul-monary oxygenation and prevent the development of BPD, especially in high-dose.     

肺表面活性物质替代治疗与支气管肺发育不良的关系

Objective To explore, the effect of different dosage of pulmonary surfactant (PS) on the inci-dence of bronchopulmanary dysplasia. Method Four hundred and three premature infants with hyaline membrane disease were divided into 3 groups according to the dose of PS: low-dose group (L-PS group, ≤ 100 mg/kg, n =188) ,high-dose group(H-PS group, > 100 mg/kg, n = 94) and no-PS group (N-PS group, n = 121). The frac-tional inspired oxygen(FiO2) and ptlmonary oxygenating function before and after 6 hours treatment were observed and the durations of oxygen therapy and mechanical ventilation, frequency of repeated intubafion, length of hospi-talization and the incidence of BPD were compared among the three groups. Results After 6 hours PS administra-tion, the FiO2,oxygen index and duration of oxygen therapy and mechanical ventilation were significantly decreased (P <0.05), while PO2 and the arterio-alveolar partial pressure of oxygen were significantly increased (P <0.05)in the H-PS and L-PS groups, compared with the N-PS group. Compared with the L-PS and N-PS groups,the H-PS group showed a decreased incidence of BPD. Conclusions PS administration could improve the pul-monary oxygenation and prevent the development of BPD, especially in high-dose.     

肺表面活性物质替代治疗与支气管肺发育不良的关系

Objective To explore, the effect of different dosage of pulmonary surfactant (PS) on the inci-dence of bronchopulmanary dysplasia. Method Four hundred and three premature infants with hyaline membrane disease were divided into 3 groups according to the dose of PS: low-dose group (L-PS group, ≤ 100 mg/kg, n =188) ,high-dose group(H-PS group, > 100 mg/kg, n = 94) and no-PS group (N-PS group, n = 121). The frac-tional inspired oxygen(FiO2) and ptlmonary oxygenating function before and after 6 hours treatment were observed and the durations of oxygen therapy and mechanical ventilation, frequency of repeated intubafion, length of hospi-talization and the incidence of BPD were compared among the three groups. Results After 6 hours PS administra-tion, the FiO2,oxygen index and duration of oxygen therapy and mechanical ventilation were significantly decreased (P <0.05), while PO2 and the arterio-alveolar partial pressure of oxygen were significantly increased (P <0.05)in the H-PS and L-PS groups, compared with the N-PS group. Compared with the L-PS and N-PS groups,the H-PS group showed a decreased incidence of BPD. Conclusions PS administration could improve the pul-monary oxygenation and prevent the development of BPD, especially in high-dose.     

T细胞亚群在轻症/重症甲型H1N1流感患者中的差异及临床意义

目的 研究T淋巴细胞亚群在不同严重程度甲型H1N1流感患者中的差异及意义.方法 收集苏州大学附属第一医院2009年9月至2010年1月收治的66例经病毒检测确诊为甲型H1N1流感患者临床资料,按疾病严重程度及预后分为轻症治愈患者(B组,47例)、重症/危重症治愈患者(C组,14例)与死亡患者(D组,5例),以同期正常人群(A组,20例)作为对照组,回顾性比较各组患者在发病后不同时间点外周血淋巴细胞计数及T淋巴细胞亚群(CD3+,CD4+,CD8+)绝对值,检测方法采用流式细胞术;并比较各组患者的发热持续时间及病毒转阴时间.采用SAS9.13统计软件进行分析,多组间均数比较采用单因素方差及SNK法.结果 甲型H1 N1流感患者在发病初期即出现淋巴细胞计数以及T细胞亚群绝对值下降,与正常人群差异具有统计学意义(P＜0.05),B、C两组随病情好转逐步上升,C组淋巴细胞计数及T细胞亚群绝对值显著低于B组(P＜0.05),且C组恢复正常的时间晚于B组,而D组则持续偏低.三组患者发热时间/病毒转阴时间分别为(4.4±1.6)vs.(4.4±1.4)d、(12.9±3.1)vs.(10.2±2.6)d、(15.2±7.3)vs.(13.3±2.9)d,三组患者之间差异具有统计学意义(P＜0.05).结论 患者感染甲型H1N1流感病毒后,导致细胞免疫功能严重受损,且外周血淋巴细胞计数及T淋巴细胞亚群的变化与疾病严重程度及预后存在紧密联系.本研究结果可以用于指导临床诊断治疗.

Abstract：

Objective To study the changes of subgroups of peripheral blood T lymphocytes in the patients infected with the 2009 pandemic influenza A ( H1N1 ) virus of different severity type. Method A total of 66 patients infected by H1N1 evidenced by RT-PCR admitted from September 2009 to January 2010 were divided into three groups: mild type ( B group, n = 47 ), cured patients of severe and critical severe type ( C group, n = 14) and died patients ( D group, n =5), according to the severity and prognosis. A total of 20 healthy volunteers served as control group( A group). Peripheral blood lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count were detected by flow cytometry at the different time points. Fever duration and H1N1 virus negative time were compared. Statistical analysis were performed by using SAS version 9.13 software and the data were processed with ANOVA and SNK test. Results Lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count declined in the early period in all the groups, and there were significant differences compared with A group (P＜0. 05), while rised with the clinical progression in group B and C,and those of C group were lower than B group ( P ＜ 0.05 ), but those of D group were always low. Fever duration and H1N1 virus negative time were (4.4 ± 1.6) days vs. (4.4 ± 1. 4) days, ( 12.9 ± 3. 1 ) days vs.( 10.2 ± 2.6) days and ( 15.2 ± 7.3 ) days vs. ( 13.3 ± 2.9 ) days respectively, and there were significant differences among the three groups ( P ＜ 0.05 ). Conclusions The cellular immune function was seriously damaged when patients were infected with H1N1. Further more, the changes of lymphocyte count, CD3+ , CD4+and CD8+ T lymphocyte count were tightly related with the degree of severity and prognosis. These findings can be used for clinical diagnosis and treatment.     

HIV 1型B/C重组病毒感染者Nef特异性T淋巴细胞免疫应答的研究

目的 研究中国主要流行的HIV 1型(HIV-1)B/C重组毒株感染者Nef蛋白特异性T淋巴细胞反应特征.方法 队列收集19例感染时间<1年和40例感染时间>3年的HIV-1 B/C重组毒株感染者,通过酶联免疫斑点试验(Elispot)检测其针对HIV-1 B/C重组毒株Nef重叠多肽产生γ干扰素(IFN-γ)的特异性T淋巴细胞反应.结果 15例感染时间<1年的HIV-1 B/C重组毒株感染者产生Nef特异性分泌IFN-γ的T淋巴细胞反应,主要识别氨基酸位置为Nef 83至135内的EVA7081.1、EVA7081.5、EVAT081.6、EVA7081.48共4条多肽;29例(75.2%)感染时间>3年的感染者产生Nef特异性分泌IFN-γ的T淋巴细胞反应,主要识别氨基酸位置为Nef 63至101内的EVA7081.43、EVA7081.44、EVA7081.45、EVA7081.47、EVA7081.48、EVA7081.49共6条多肽.感染时间<1年的感染者平均反应强度为284.13 SFC/106PBMC,感染时间>3年的感染者平均反应强度为152.44 SFC/106PBMC,2组比较差异具有统计学意义(U=91.000,P=0.002).结论 HIV-1 B/C重组病毒感染者在感染时间<1年和>3年时机体均识别HIV-1 Nef的中心区域,其产生IFN-γ的特异性T淋巴细胞反应强度随疾病的进展逐渐减小.     

Impact of human immunodeficiency virus type 1 subtype on first-line antiretroviral therapy effectiveness

OBJECTIVE: The effectiveness of antiretroviral treatment (ART) was compared in 416 naive patients from a French clinical cohort infected with B and non-B HIV-1 subtypes. METHODS: Time to HIV viral load (VL) undetectability was calculated for each subtype group. Three other parameters were estimated 3, 6 and 12 months after enrolment: clinical progression (that is, AIDS-defining events or death), changes in CD4 cell counts from baseline and proportion of patients achieving an undetectable VL (<400 HIV-RNA copies/ml). RESULTS: In this cohort, 317 patients (76%) were infected with a B subtype and 99 (24%) with a non-B subtype. Median time to VL undetectability was similar in the B subtype group [147 days, 95% confidence interval (CI) 119-165] and non-B subtype group (168 days, 95% CI: 105-234; P=0.16). After adjusting for AIDS-defining events at enrolment, baseline CD4 cell counts and VL, and for the treatment on which patients were initiated, no association was found between HIV subtypes and time to VL undetectability (B subtype vs non-B subtype: hazard ratio=0.80, 95% CI: 0.62-1.02, P=0.07). In the 3, 6 and 12 months after enrolment, subtype had no impact on clinical progression, CD4 cell count or VL responses to ART. This suggests that B and non-B subtypes do not affect first-line therapy efficacy, which is encouraging in view of the worldwide spread of non-B HIV-1 subtypes and the increasing availability of ART in developing countries. However, in this study we did not take into account individual non-B subtype species, therefore further studies should be designed to evaluate the efficacy of these regimens in patients with particular non-B subtypes.     

Comparative clonal analysis of human immunodeficiency virus type 1 (HIV- 1)-specific CD4+ and CD8+ cytolytic T lymphocytes isolated from seronegative humans immunized with candidate HIV-1 vaccines

The lysis of infected host cells by virus-specific cytolytic T lymphocytes (CTL) is an important factor in host resistance to viral infection. An optimal vaccine against human immunodeficiency virus type 1 (HIV-1) would elicit virus-specific CTL as well as neutralizing antibodies. The induction by a vaccine of HIV-1-specific CD8+ CTL in humans has not been previously reported. In this study, CTL responses were evaluated in HIV-1-seronegative human volunteers participating in a phase I acquired immune deficiency syndrome (AIDS) vaccine trial involving a novel vaccine regimen. Volunteers received an initial immunization with a live recombinant vaccinia virus vector carrying the HIV-1 env gene and a subsequent boost with purified env protein. An exceptionally strong env-specific CTL response was detected in one of two vaccine recipients, while modest but significant env-specific CTL activity was present in the second vaccinee. Cloning of the responding CTL gave both CD4+ and CD8+ env-specific CTL clones, permitting a detailed comparison of critical functional properties of these two types of CTL. In particular, the potential antiviral effects of these CTL were evaluated in an in vitro system involving HIV-1 infection of cultures of normal autologous CD4+ lymphoblasts. At extremely low effector-to-target ratios, vaccine-induced CD8+ CTL clones lysed productively infected cells present within these cultures. When tested for lytic activity against target cells expressing the HIV-1 env gene, CD8+ CTL were 3-10-fold more active on a per cell basis than CD4+ CTL. However, when tested against autologous CD4+ lymphoblasts acutely infected with HIV-1, CD4+ clones lysed a much higher fraction of the target cell population than did CD8+ CTL. CD4+ CTL were shown to recognize not only the infected cells within these acutely infected cultures but also noninfected CD4+ T cells that had passively taken up gp120 shed from infected cells and/or free virions. These results were confirmed in studies in which CD4+ lymphoblasts were exposed to recombinant gp120 and used as targets for gp120-specific CD4+ and CD8+ CTL clones. gp120-pulsed, noninfected targets were lysed in an antigen-specific fashion by CD4+ but not CD8+ CTL clones. Taken together, these observations demonstrate that in an in vitro HIV-1 infection, sufficient amounts of gp120 antigen are produced and shed by infected cells to enable uptake by cells that are not yet infected, resulting in the lysis of these noninfected cells by gp120-specific, CD4+ CTL.(ABSTRACT TRUNCATED AT 400 WORDS)     

HIV-1-infected children on HAART: immunologic features of three different levels of viral suppression

BACKGROUND: HIV-1-infected children show changes of blood lymphocyte subpopulations. We have, therefore, investigated how highly active anti-retroviral therapy (ART) alter these subsets. Blood samples were taken from 41 HIV-1-infected children on ART who were divided into groups showing good, partial and poor responses to ART on the basis of viral load (VL) measurement in blood. The observations were compared to those seen in 20 uninfected children. METHODS: The samples were studied using 4-color flow cytometry for "na?ve", central memory and effector memory cells as well as for CD38 expression as the sign of activation within both the CD4+ and the CD8+ T cell populations. HIV-1 infected children were also evaluated for the presence and the titers of antibodies induced by vaccination against childhood infections in our patients while on HAART. RESULTS: Lymphocyte counts were lower in the "poor" viral load responding (VLR) group when compared with partial and good VLRs. Poor VLRs had lower total and na?ve CD4+ T cell counts. HIV-1-infected children from all three groups had high CD8+ T cell counts. Central memory CD4+ and CD8+ T cell percentages were particularly low in the poor VLR group while in the poor VLR group the percentages of effector memory CD4+ and CD8+ T cells were higher when compared with the control group. Higher cellular activation of CD8+ T cells was observed in HIV-1-infected children, particularly when analyzed for the intensity of CD38 expression in the poor VLR group. CD5 expression on B cells was higher among all HIV-1-infected children. Antibodies to tetanus, diphtheria, measles, rubella, and hepatitis B were present in a large proportion of children but the titers were similarly low for all three groups of HIV-infected children. CONCLUSIONS: Children with different levels of viral response to HAART present immune phenotype characteristics that tend to place the children with partial and good virological responses into the same group. These children are still moderately deficient in their immune responses but show better recovery than seen with children in the poor VLR group. These observations indicate that the proportions of central memory cells among the CD4+ T cells and the intensity of the expression of CD38 activation antigen on CD8+ T cells provide more informative parameters for monitoring children on HAART than the absolute numbers of CD4+ and CD8+ T cells alone.     

Expansion of pre-existing, lymph node-localized CD8+ T cells during supervised treatment interruptions in chronic HIV-1 infection

To date, most studies have focused on the characterization of HIV-1-specific cellular immune responses in the peripheral blood (PB) of infected individuals. Much less is known about the comparative magnitude and breadth of responses in the lymphoid tissue. This study analyzed HIV-1-specific CD8+ T cell responses simultaneously in PB and lymph nodes (LNs) of persons with chronic HIV-1 infection and assessed the dynamics of these responses during antiretroviral treatment and supervised treatment interruption (STI). In untreated chronic infection, the magnitude of epitope-specific CD8+ T cell activity was significantly higher in LNs than in PB. Responses decreased in both compartments during highly active antiretroviral therapy, but this decline was more pronounced in PB. During STI, HIV-1-specific CD8+ T cell responses in PB increased significantly. Enhancement in breadth and magnitude was largely due to the expansion of pre-existing responses in the LNs, with new epitopes infrequently targeted. Taken together, these data demonstrate that HIV-1-specific CD8+ T cells are preferentially located in the LNs, with a subset of responses exclusively detectable in this compartment. Furthermore, the enhanced CD8+ T cell responses observed during STI in chronically infected individuals is largely due to expansion of pre-existing virus-specific CD8+ T cells, rather than the induction of novel responses.     

HIV/AIDS患者血常规检测结果与CD4+T淋巴细胞计数之间的相关性研究

目的 通过对HIV感染者和AIDS患者血常规总淋巴细胞数(TLC)、Hb、PLT、WBC与CD4+ T淋巴细胞计数相关性的研究,探讨用血常规多项指标检测联合预测HIV/AIDS患者CD4+ T淋巴细胞计数的可行性.方法 778例HIV/AIDS患者共采集1 038份血样,血常规中四项指标:TLC、Hb、WBC、PLT与CD4+T淋巴细胞计数相关分析采用Spearman秩和相关.绘制受试者工作特征(ROC)曲线以判断各项指标预测CD4+T淋巴细胞计数的真实度及其最佳临界值,计算各临界值的敏感度、特异度、阳性预测值和阴性预测值.采用联合试验评价多指标联合预测CD4+ T淋巴细胞计数<200个/μl的可行性.结果 TLC、Hb、WBC、PLT与CD4+T淋巴细胞计数之间均存在正相关,相关系数分别为r=0.64,P=0.000;r=0.36,P=0.000;r=0.24,P=0.000;r=0.09,P=0.000.TLC、Hb预测CD4+ T淋巴细胞计数的ROC曲线下面积分别在0.82～0.84、0.66～0.70之间.单独使用TLC预测CD4+ T淋巴细胞计数<50、200、350个/μl的最佳临界值分别为TLC<1 100 × 106/L、1 200 ×106/L、1 400 × 106/L.TLC<1 200 × 106/L与Hb<120 g/L联合预测CD4+ T淋巴细胞计数<200个/μl的敏感度为45.3%,特异度为82.8%.结论本研究结果显示TLC<1 200 ×106/L与Hb<120g/L联合预测CD4+ T淋巴细胞计数<200个/μl的临床使用价值不高.     

高容量血液滤过对多器官功能障碍犬内毒素及CD14的影响

目的 观察高容量血液滤过(HVHF)治疗多脏器功能障碍综合征(MODS)犬中血浆内毒素(LPS)及其受体CD14的变化,探讨HVHF防治MODS的作用机制。方法 12只Beagle 犬采用失血性休克+复苏灌注+内毒素血症建立MODS模型,随机(随机数字法)分为HVHF组(n=6)和MODS组(n=6),HVHF组在LPS注射完毕后给于HVHF治疗24h。分别于术前(T1)、LPS注射前(T2)、LPS注射完后0 h(T3),3 h(T4),6 h(T5),9 h(T6),12 h(T7),15 h(T8)及24 h(T9),观察血浆LPS及其受体CD14、器官功能指标变化。LPS采用MB-80微生物快速动态检测系统测定。CD14采用ELISA测定。统计学方法各时间点前后采用重复测量资料方差分析。结果 HVHF治疗后血浆LPS及CD14明显下降,在T4-9时间点明显低于MODS组(P＜0.01)。超滤液中能检测出CD14,SC为(0.81±0.09)。超滤液中未能检测出LPS。HVHF组主要器官功能明显改善,平均动脉压在T4-9时间点显著高于MODS组(P＜0.0l)。动脉血氧分压逐渐升高,与MODS组在T4-9时间点比较差异有统计学意义(P＜0.01)。HVHF治疗后肌酐、尿素氮逐渐下降,与MODS组在T4-9时间点比较差异有统计学意义(P＜0.0l)。结论 HVHF能改善主要器官功能、降低血浆LPS及CD14峰值,有效遏制过度炎症反应。     

60Coγ射线全身照射对小鼠胸腺中叉状头转录因子N1表达水平的影响

目的 探讨60Co γ射线全身照射(TBI)诱导的小鼠胸腺损伤,对叉状头转录因子N(Foxn)1及其相关基因表达的影响,以及该变化水平与胸腺损伤程度的相关关系.方法 选择65只雄性C57BL/6小鼠作为研究对象.其纳入标准:无特殊病原体(SPF)级,6～8周龄,体重为18.0～21.0 g.采用简单随机法将其分为4组;①致死剂量全身照射(L-TBI)5 d组(n=10),小鼠接受剂量为7.5Gy的60Coγ射线TBI;②非致死剂量全身照射(SL-TBI)5 d组(n=10),小鼠接受剂量为5.5 Gy的TBI;③SL-TBI 24 d组,小鼠接受剂量为5.5 Gy的TBI(n=10);④对照组(n=5),小鼠不进行任何处理.L-TBI 5 d组、SL-TBI 5 d组及对照组小鼠均在TBI处理5d后被处死,获取胸腺组织;L-TBI 24 d组小鼠于24 d后被处死,获取胸腺组织.剩余30只小鼠用于梯度剂量60Co γ射线照射实验.采用简单随机法将其按照接受60 Coγ射线TBI剂量平均分为6组,每组5只小鼠,即0 Gy剂量组、1.0 Gy剂量组、2.0Gy剂量组、3.0 Gy剂量组、4.0 Gy剂量组及5.0 Gy剂量组.6组小鼠在TBI处理5d后处死,获取胸腺组织.本研究中TBI剂量率均为0.5 Gy/min.采用流式细胞术(FCM)分析上述10组小鼠胸腺各细胞群的细胞数变化.应用定量聚合酶链反应(PCR)技术检测胸腺中与胸腺功能相关的转录因子Foxn1及相关基因mRNA相对表达水平的变化.对小鼠胸腺各细胞群的细胞数,Foxn1及相关基因mRNA相对表达水平进行统计学分析;对Foxn1与白细胞介素(IL)-22 mRNA相对表达水平的相关性、Foxn1 mRNA相对表达水平与胸腺各细胞群细胞数的相关性进行统计学分析.结果 ①L-TBI 5 d组、SL-TBI 5 d组、SL-TBI 24 d组及对照组小鼠胸腺组织中总细胞、双阳性(DP)胸腺细胞、CD4+单阳性(SP)胸腺细胞、CD8+ SP胸腺细胞、双阴性(DN)2、DN3胸腺细胞及胸腺上皮细胞(TEC)数相比,差异均有统计学意义[平均总细胞数:(32.3±6.1)×106、(39.2±8.1)×106、(187.2±20.3)×106、(282.5±20.3)×106个,DP胸腺细胞数中位数:4.1×106、0.5×106、123.3×106、240.3×106个,CD4+ SP胸腺细胞数中位数:5.2×106、10.4×106、26.8×106、20.5×106个,CD8+ SP胸腺细胞数中位数:1.9×106、2.4×106、10.2×106、6.5×106个;F/x2=310.471、17.863、16.352、16.419,P=0.000、0.000、0.001、0.001];但4组小鼠的DN2与DN3胸腺细胞及TEC数相比,差异却无统计学意义(DN2与DN3胸腺细胞数中位数:2.2×106、9.8×106、4.8×106、8.1×106个,TEC数中位数:1.9×106、4.6×106、2.5×106、3.3×106个;x2=7.574、7.241,P=0.056、0.065).与对照组相比,L-TBI 5 d组、SL-TBI 5 d组小鼠胸腺组织中总细胞数均减低,且差异有统计学意义(P=0.000、0.000).其中,L-TBI 5 d组、SL-TBI 5 d组小鼠DP胸腺细胞数显著低于对照组,且差异亦有统计学意义(P=0.045、0.001).与SL-TBI 5 d组相比,SL-TBI 24 d组小鼠胸腺组织中总细胞数与DP胸腺细胞数均显著增高,且差异有统计学意义(P=0.000、0.045).②L-TBI 5 d组、SL-TBI 5 d组、SL-TBI 24 d组及对照组小鼠胸腺组织中Foxn1与IL-22 mRNA相对表达水平相比,差异均有统计学意义(Foxn1mRNA相对表达水平:15.2±1.2、10.8±1.1、6.6±0.8、1.0±0.1,IL-22 mRNA相对表达水平:11.2±0.8、7.2±1.2、4.4±0.5、1.0±0.1;F=233.971、161.067,P=0.000、0.000).与对照组相比,L-TBI 5 d组、SL-TBI 5 d组小鼠胸腺中Foxn1mRNA相对表达水平显著增高(P=0.000、0.000);同时,这两组小鼠胸腺中IL-22 mRNA相对表达水平亦显著增高(P=0.000、0.000).与SL-TBI 5 d组相比,SL-TBI 24 d组小鼠胸腺中Foxn1与IL-22 mRNA相对表达水平显著减低(P=0.000、0.000).小鼠胸腺组织中Foxn1与IL-22 mRNA相对表达水平存在正相关关系(r=0.976,P＜0.05).③梯度剂量TBI处理后,6个剂量组胸腺组织中Foxn1 mRNA相对表达水平与DP胸腺细胞数存在负相关关系(r=-0.930,P＜0.05).④L-TBI 5 d组、SL-TBI 5 d组及对照组的CC趋化因子配体(CCL)25、delta样配体(DLL)4、自身免疫调节蛋白(Aire)与骨形成蛋白(BMP)4 mRNA相对表达水平相比,差异均有统计学意义(CCL25 mRNA相对表达水平:11.2±1.5、11.4±1.2、1.0±0.1,DLL4 mRNA相对表达水平:7.3±0.9、6.3±0.3、1.0±0.1,Aire mRNA相对表达水平:5.1±1.0、5.0±0.1、1.0±0.1,BMP4 mRNA相对表达水平:4.4±1.4、3.6±0.5、1.0±0.1;F=148.862、197.667、73.911、21.471,P=0.000、0.000、0.000、0.000);3组中Foxn1上游基因同源框蛋白(Hox)a3 mRNA相对表达水平相比,差异却无统计学意义(1.4±0.4、0.8±0.3、1.0±0.1相比;F=5.368,P=0.221).与对照组相比,L-TBI 5 d组中Foxn1下游基因CCL25、DLL4与Aire的mRNA相对表达水平均显著增高(p=0.000、0.000、0.000);SL-TBI 5 d组中CCL25、DLL4与Aire mRNA相对表达水平亦显著增高(P=0.000、0.000、0.000).同时,与对照组相比,L-TBI 5 d组、SL-TBI 5 d组中BMP4mRNA相对表达水平存在一定程度的增高,且差异亦有统计学意义(P=0.000、0.000).结论 60C0γ射线TBI可导致小鼠胸腺中Foxn1mRNA表达水平增高,Foxn1表达水平上调及其相关基因表达水平的变化与胸腺受损程度的关系密切.     

儿科不同年龄组呼吸道病毒感染IgM含量与免疫功能的相关性

目的 探讨儿科不同年龄组呼吸道病毒感染者血清IgM含量、病毒IgM检出率与体液、细胞免疫的关系.方法 患儿按年龄分为3组：1组,3个月至1岁,42例；2组,1～3岁,38例；3组,3～ 12岁,40例.免疫比浊法检测患儿血清IgM含量,间接免疫荧光法检测病毒IgM阳性率,流式细胞术检测CD3、CD4、CD8、NK及B淋巴细胞.结果 血清IgM含量及病毒IgM检出率1组与2、3组差异具有统计学意义（P＜0.05）,血清IgM含量与病毒IgM检出率呈正相关（r=0.998,P＜0.05）.按百分率统计1组与2、3组间CD4,1、3组间CD8,NK、B细胞各组间差异具有统计学意义（P＜0.05）.按绝对数统计,三组间CD3、CD4及B细胞,1、3组间CD8,1组与2、3组间CD4/CD8差异有统计学意义（P＜0.05）,血清IgM含量与CD3、CD4、CD8及B细胞绝对数呈负相关（r=-0.966,P＜0.05；r=-0.989,P＜0.05；r=-0.911,P＜0.05；r=-0.964,P＜0.05）.结论 低年龄组免疫细胞绝对数高于高年龄组,但产生IgM水平低,致检出率低,提示低龄组免疫功能不健全.