红细胞天然免疫分子CD59与可溶性IL-2R相关性分析

为了探讨红细胞CD5 9与可溶性IL 2受体 (sIL 2R )之间变化规律。采用流式细胞仪测定红细胞CD5 9,用ELISA法测定sIL 2R含量。结果发现肿瘤患者红细胞CD5 9数量 (平均荧光相对强度 5 2 5 8± 12 4 2 )明显低于 2 6例银屑病患者 ( 6 31 0 0±16 6 70 ,P <0 0 5 ) ,而 19例正常人sIL 2R含量 ( 1736 5 8± 6 5 0 10 )ng/L明显低于肿瘤患者 ( 2 36 4 80± 831 2 0 ,P <0 0 5 ) ,6 5例标本同时测定红细胞CD5 9和sIL 2R ,两者之间存在负相关 (r= 0 312 ,P <0 0 5 )。红细胞CD5 9与sIL 2R之间变化的分析为免疫反应的调控规律及临床提供了有用的信息     

系统性红斑狼疮患者外周血淋巴细胞CD1c的表达

目的 :研究系统性红斑狼疮 (SLE)患者外周血淋血细胞CD1c的表达情况及与疾病活动性之间的关系。方法 :用流式细胞仪检测了 4 7例SLE患者外周血淋巴细胞CD1c的表达及淋巴细胞表型分析 ,并评价与疾病活动性的关系。结果 :SLE活动组病人CD1c 细胞百分率显著增高 (P <0 0 5 ) ,CD4 细胞百分率显著降低 (P <0 0 1) ,CD3 、CD8 细胞百分率正常 ,CD2 0 细胞数增高 (P <0 0 1)。稳定期病人CD1c 细胞百分率正常 ,CD4 、CD8 、CD2 0 细胞百分率均正常。SLE患者CD1c细胞阳性率与患者SLEDAI的评分有显著的相关性 (r=0 6 8,P <0 0 1) ,与抗dsDNA抗体的表达有显著相关性 (r =0 36 ,P <0 0 5 ) ,与抗心磷脂抗体的表达有显著的相关性 (r=0 6 4 ,P <0 0 1) ,与血清C3水平有显著相关性 (r =- 0 35 ,P <0 0 5 )。活动期病人经治疗后CD1c的表达明显下降。结论 :系统性红斑狼疮患者外周血CD1c表达与疾病的活动性明显相关 ,CD1c可能在SLE脂类抗原及核酸类抗原的递呈及抗双链DNA抗体、抗磷脂抗体的产生中起重要作用。     

2型糖尿病患者免疫功能变化与血糖和胰岛素水平相关性的研究

探讨2型糖尿病患者免疫功能变化及其与血糖和胰岛素的相关性.检测34例2型糖尿病(T2DM)患者在空腹和口服75克葡萄糖后120min时的红细胞C3b受体花环(RBC-C3bRR)、红细胞免疫复合物花环(RBC-ICR)、T淋巴细胞亚群(CD3、CD4、CD8、CD4/CD8)、可溶性白介素-2受体(sIL-2R)、免疫球蛋白(IgG、IgA、IgM)、血糖和胰岛素.对照组30名.2型糖尿病患者与对照组相比,RBC-C3bRR、CD3、CD4、CD4/CD8和IgG、IgA均明显降低(P＜0.01,P＜0.05),而RBC-ICR、sIL-2R和CD8明显增高(P＜0.01,P＜0.05),服葡萄糖后120min时较空腹时更明显.RBC-C3bRR与血糖和胰岛素呈负相关(r1=-0.354,r2=-0.335,P＜0.05);RBC-ICR与血糖和胰岛素呈正相关(r3=0.368,r4=0.342,P＜0.05);CD3、CD4、CD4/CD8、IgA与血糖呈负相关(r5=-0.302,r6=-0.378,r7=-0.413,P＜0.01;r8=-0.332, P＜0.01);CD8和sIL-2R与血糖呈正相关(r9=0.214,P＜0.05;r10=0.437,P＜0.01).T2DM患者存在多项免疫指标变化,其免疫功能低下与血糖和胰岛素升高有相关性.     

高通量血液透析对维持性血液透析患者细胞免疫功能的影响

目的探讨高通量血液透析对维持性血液透析（MHD）患者细胞免疫功能的影响。方法收集2012年3月至8月于本院门诊行MHD治疗的患者40例,随机数字表法分为血液透析（HD）组（n=20）和高通量血液透析（HFHD）组（n=20）,分别接受HD和HFHD治疗,均为每周透析3次,每次4h。透析前、透析后4、24、48h,流式细胞术检测两组患者外周血CD4＋．CD8＋、CD25＋,记录CD47CD8＋比值,酶联免疫吸附测定（ELISA）检测血清IL-2、可溶性IL-2受体（sIL．2R）;另设健康对照组（C组）20例,清晨空腹抽血检测上述指标。结果与C组比较,透析前HD组和HFHD组患者外周血CD4＋、CD25＋、CD4＋／CD8＋水平下降,血清IL．2水平下降,sIL．2R升高（均P〈0．05）。与透析前比较,HD组患者透析后4h外周血CD4＋、CD25＋、CD47CD8＋水平升高,血清IL-2水平升高,sIL-2R降低（均P〈0．05）,CD8＋差异无统计学意义（P〉O．05）;与透析前比较,HD组患者透析后24、48h上述各指标差异无统计学意义（均P〉0．05）。与透析前比较,HFHD组患者透析后4、24、48h外周血CD4＋、CD25＋、CD4VCD8＋水平升高,血清IL．2水平升高,slL-2R降低（均P〈0．05）,而CD8＋差异均无统计学意义（均P〉O．05）。与同时点HD组比较,HFHD组透析后4h各指标差异均无统计学意义（均P〉O．05）;透析后24、48h,HFHD组外周血CD4＋、CD25＋、CD4＋／CD8＋水平升高,血清IL．2水平升高,sIL．2R降低[CD4＋：（38．73±6．25）％比（34．92±5．84）％,（37．03±5．41）％比（32．62±5．79）％;CD25＋：（21．36±4．65）％比（15．29±4．72）％,（18．19±4．27）％比（13．94±5．05）％;CD4＋／CD8＋：1．42±0．31比1．23±0．29,1．38±0．30比1．20±0．33;IL-2：（22．03±5．18）m±L比（19．03±4．87）m#L,（20．54±5．92）mL比（18．26±4．96）mL;sIL-2R：（672．96±159．36）U／ml比（787．32±143．27）u,ml,（720．24±143．92）u,（858,42±172．13）U／ml,均P〈0．05],而CD8＋差异无统计学意义（均P〉O．05）。结论HD可短暂改善MHD患者的细胞免疫功能,HFHD可持续改善MHD患者的细胞免疫功能。     

系膜增生性肾炎患者血清IL-4、sIL-6R、IL-10的检测及其临床意义

了解系膜增生性肾炎 (MsPGN )患者血清白细胞介素 4(IL 4)、白细胞介素 10 (IL 10 )和可溶性白细胞介素 6受体 (sIL 6R )水平及其意义。用双抗体夹心ELISA法同步定量测定经肾脏病理活检确诊的 44例MsPGN患者及正常人 40例血清IL 4、sIL 6R及IL 10的水平。MsPGN患者血清中的IL 4、IL 10及sIL 6R明显高于正常人 (P <0 0 1) ;在MsPGN中临床符合原发性肾病综合征 (PNS )的患者血清IL 4水平与其 2 4h尿蛋白定量 (2 4hUPQ )呈正相关 (n =31,r=0 5 0 1,P <0 0 1)。在MsPGN患者的发病机制中 ,IL 4和sIL 6R作为一组致炎症因子介导肾炎的免疫反应过程 ,引起机体的免疫损伤 ;而IL 10则为抑炎症因子 ,对机体可能有保护作用     

不稳定型心绞痛病人血清sFas、sFasL及sIL-2R水平的临床价值

目的　探讨血清sFas、sFasL和sIL 2R水平与不稳定型心绞痛 (Unstableanginapectoris,UAP)之间的关系。方法　应用酶联免疫吸附双抗体夹心 (ELISA)法 ,测定了 32例UAP病人 (UAP组 )和 2 0例对照组受试者血清sFas、sFasL和sIL 2R水平。结果　UAP组病人血清sFas和sIL 2R水平均明显高于对照组 (P <0 .0 1) ,而 2组sFasL水平无显著性差别 (P >0 .0 5 )。UAP组病人sFas水平与sIL 2R水平存在显著正相关 (r=0 .4 4 7,P <0 .0 5 )。结论　高水平的血清sFas、sIL 2R与UAP有关。高水平的血清sFas可能通过维持自身免疫炎性反应而导致UAP的发生发展     

Graves病患者外周血细胞间粘附分子-1水平检测的临床价值

目的 :探讨外周血细胞间粘附分子 1(ICAM 1)水平检测在Graves病患者中的价值。方法 :用流式细胞术测定 37例Graves病 (GD)患者治疗前外周血淋巴细胞膜上细胞间粘附分子 1(CD5 4 )、CD4 、CD3 、CD8 、CD19 、CD2 5 (可溶性白介素 2受体 )的表达水平 ,用ELISA方法测定其血清中可溶性细胞间粘附分子 1(sICAM 1)的浓度 ,并测定其它体液免疫指标(TsAb、TGAb、TpoAb) ,同时和 2 2例正常对照人群进行各指标间的比较 ;Graves病患者TsAb阳性率与sICAM 1阳性率进行比较 ;此外 ,在细胞间粘附分子 1水平与机体其它免疫指标间做相关分析。结果 :Graves病患者血清可溶性细胞间粘附分子 1水平明显高于对照人群 (P <0 0 0 1) ,但与体液免疫指标间无相关关系 ,与细胞免疫指标中的CD4 水平呈负相关 (n =37,rs =- 0 4 91,P<0 0 0 5 ) ;GD病患者sICAM 1的阳性率 (89 19% )显著高于TsAb的阳性率 (6 7 5 7% ,P <0 0 5 )。结论 :sICAM 1在未治疗的GD病患中升高 ,其阳性率显著高于TsAb的阳性率 ,其升高与体液免疫指标不相关 ,与细胞免疫指标有一定关系     

结核病患者外周血T细胞亚群、mIL-2R与sIL-2R的测定

采用生物素 链霉亲和素 (BSA)系统检测 85例结核病患者T细胞亚群及经PHA诱导前后其mIL 2R表达水平 ,用ELISA法测定其血清sIL 2R水平 ,同时选取健康献血员 2 5名为正常对照 ,操作按说明书进行。资料分析采用t检验。从表 1可见 :结核病患者外周血CD3+ 、CD4 + 百分率、CD4 + CD8+ 比值均低于正常对照 (P <0 .0 1) ;CD8+ 百分率、　　表 1　结核病患者与正常对照组中外周血T细胞亚群、PBMC经PHA诱导前后mIL 2R表达水平和血清sIL 2R检测结果[n , x±s ;百分率 (% ) ]组别n CD3 + ( % )CD4+ ( % )CD8+ ( % )CD4+ CD8+sI…     

淫羊藿对血液透析HCV感染者细胞免疫功能的作用

为探讨丙型肝炎病毒 (HCV)感染对维持性血液透析 (血透 )患者细胞免疫功能的影响和淫羊藿对其免疫调节作用 ,应用ELISA和RT PCR检测 6 2例血透患者抗HCV和HCVRNA ,ELISA法检测患者血清sIL 2R、IL 6和TNF α水平 ,同时观察应用中药淫羊藿治疗同一血透患者 3个月前后上述指标的变化。结果表明 ,6 2例血透患者中抗HCVIgM阳性 2 7例 (4 3 6 % ) ,抗HCVIgG阳性 2 9例 (4 6 8% ) ,HCVRNA阳性 34例 (5 4 8% ) ,至少一项阳性 37例 (5 9 7% ) ;与血透患者HCV标志阴性组相比 ,阳性组血清sIL 2R、IL 6和TNF α水平显著增高 ;淫羊藿治疗后不论HCV阳性与否此三项指标均显著降低。结果提示 ,血透患者均存在明显的细胞免疫功能紊乱 ,感染HCV后免疫功能更低下 ,淫羊藿有显著调节血透患者细胞免疫功能的作用。     

原发性肝癌患者CTCs与CD4~+/CD8~+、NLR、TTV及肿瘤分期的相关性研究

目的:探讨原发性肝癌患者外周血循环肿瘤细胞(CTCs)与T细胞亚群CD4~+/CD8~+比值、中性粒细胞与淋巴细胞比值(NLR)、肿瘤总体积(TTV)及肿瘤分期的关系。方法:收集80例原发性肝癌患者外周血,采用益善生物技术公司使用的"免疫去除结合纳米过滤法"Can PatrolTM行CTCs检测。同时使用流式细胞术检测外周血中CD4+T细胞和CD8~+T细胞数量;收集患者外周血中性粒细胞数、淋巴细胞数、肿瘤最大直径及肿瘤分期等临床病理参数。计算出外周血CD4~+/CD8~+比值、NLR及TTV。根据CD4~+/CD8~+平均值及TTV、NLR中位数将CD4+/CD8+比值、TTV和NLR分为高、低2组;根据肝癌第7版TMN分期标准将患者分为I+II期和III+IV期2组。分别比较CTCs在2组中的差别。结果:高CD4~+/CD8~+比值组外周血CTCs数及间质型CTCs数明显低于低CD4~+/CD8~+比值组(P0.05)。高TTV组外周血的CTCs数、间质型CTCs数及混合型CTCs数明显高于低TTV组(P0.05);肿瘤分期I、II期组外周血的CTCs数、间质型CTCs数及混合型CTCs数明显低于III、IV期组(P0.05)。高NLR组与低NLR组外周血的CTCs数、间质型CTCs数、混合型CTCs数和上皮型CTCs数间的差异无统计学显著性。结论:原发性肝癌患者外周血中存在CTCs;外周血CTCs与T细胞免疫、TTV及肿瘤分期有显著相关:T细胞免疫越差、TTV越大及肿瘤分期越晚,外周血的CTCs越多。     

328例血清TT病毒DNA及其IgG抗体的检测

目的 了解不同人群中TT病毒（TTV）感染情况。方法 根据Okamoto报道的TTV全序列设计引物,建立TTV DNA套式聚合酶链反应,利用该法对81例正常人、92例献血员、123例甲－庚型肝炎,32例非甲－庚型肝炎病人进行TTV DNA检测,同时用ELISA法检测抗TTV IgG。结果 TTV在以上四种人群中阳性率分别为2．5％、2．2％、19．5％、28．1％,抗TTV IgG的阳性率分别为1．2％、3．7％、26．8％、34．4％。前者与后者两者比较差异存在显著性（P＜0．05）;重叠感染中TTV合并HBV的二重感染率最高为75．0％。结论 不同人群匀存在TTV感染;正常人群和职业献血员存在健康携带状态;甲－庚型肝炎和非甲－肝炎病人为高危人群;TTV可与各型肝炎存在重叠感染;TTV除经血传播外,存在其他传播途径,抗TTV IgG可作为TT病毒感染的检测手段。     

Relationships between total plasma load of torquetenovirus (TTV) and TTV genogroups carried

In 239 torquetenovirus-positive people, multiple-genogroup infections were common and associated with higher viral loads than would be expected from simple additive effects. The latter observation was restricted to the infections which included both genogroups 1 and 3, pointing to the possible existence of some kind of infection facilitation between these genogroups.     

贵州地区不同人群TTV核酸检测及部分核苷酸序列分析

目的 了解贵州地区TTV感染状况，分析TTV贵州株的基因特点。方法 以公布的TTV第1读码区序列设计两对寡核苷酸引物，用套式聚合酶链反应法(nested—PCR)检测贵州地区不同人群血清中TTV核酸(TTV DNA)，并对3份TTV DNA阳性血清的PcR产物，用直接测序法测定核苷酸序列。结果 62例正常人，37例志愿献血员，50例血液透析患者，107例静脉药瘾者及139例肝病患者血清中TTV DNA阳性率分别为6．45％(4／62)，8．1％(3／37)，26.0％(13／50)，25.23％(27／107)和16.55％(23／139)。在肝病组中，重型肝炎、肝硬化、肝癌患者的TTV DNA阳性率分别为35.71％(5／14)，14．15％(15／106)和15．79(3／19)。测定的282个核苷酸中，3株贵州株的同源性均高于99％，与日本株N22相比，同源性都为98％。结论 贵州存在TTV感染，血透患者中有较高的TTV感染率，TTV可能是重型肝炎的病原因子，3株贵州株可能为同一基因型。     

Highly diverse TTV population in infants and their mothers

Infants born to serum HCV-positive 12 mothers were enrolled in the study. Nucleotide sequences amplified by primers deduced from a noncoding region were compared between mothers and their infants. The rates for detection of serum TTV in 12 mothers and their infants were 10/12 (83%) and 9/12 (75%), respectively. Serum TTV DNA was not detected in any infant at 1 month of age, but was detected for the first time between 1.5 and 8 months after birth. Positivity persisted thereafter throughout the follow-up period. In seven randomly selected mother-infant pairs, intrahost TTV heterogeneity was lower in infants than in mothers. Furthermore, one of seven mother-infant pairs showed a high degree of similarity (98.7-100%) in all clones, while in four infants, all nucleotide sequences differed by >10% from those of their mothers. However, the degree of homology in the two mother-infant pairs was 89-98.7% in family 2 and 88.1-99.4% in family 5. In the present study, with only one exception, it was shown that TTV from infants is not identical to TTV from mothers. The mechanism is discussed briefly in this paper.     

TTV infection in children born to mothers infected with TTV but not with HBV, HCV, or HIV

The TT virus (TTV) was isolated recently from the serum of a patient with post-transfusion hepatitis. TTV infection is widespread in the general population, and its prevalence increases continuously with age. The pathogenic role of TTV in liver disease remains controversial, and the source of transmission is still unclear. We investigated the pathogenicity and epidemiology of TTV infection in infants born to TTV DNA-positive mothers. Enrolled in this study were 22 mother-child pairs testing negative for antibodies to hepatitis B, hepatitis C, and the human immunodeficiency viruses (HIVs). The children were followed for 30 months after birth. Serum TTV DNA was detected by N22-PCR, and the PCR products were cloned and sequenced. The prevalence of TTV infection in children increased with age. Of the 22 children, 13 (59%) became positive for TTV DNA during the follow-up period. Of these 13 children, 6 (46%) had elevated levels of serum alanine aminotransferase (ALT), although the elevations were transient and mild. TTV viremia was not associated significantly with the abnormal ALT levels. Children with TTV viremia developed neither severe liver disease nor fulminant hepatitis. Phylogenetic analysis showed that, in 11 (85%) of the 13 pairs, the mother and child had the same genotype at the first PCR-positive time point. Among those 11 mother-child pairs, 6 (55%) had identical TTV nucleotide sequences. However, the genotype of predominant clones changed in 5 (50%) of 10 children who were positive for TTV DNA at two or more time points during the follow-up period. In conclusion, this study did not provide evidence that TTV infection is related to liver disease in children. Although the main source of TTV infection in children is presumed to be their mothers, transmitted via non-parenteral routes in the course of daily contact, intrafamilial carriers may also be sources of TTV infection.     

TT virus (TTV) loads associated with different peripheral blood cell types and evidence for TTV replication in activated mononuclear cells

TT virus (TTV) loads associated with the peripheral blood cells of seven patients known to carry the virus in plasma were investigated by real-time PCR. Whereas red cells/platelets were uniformly negative, six and four patients yielded positive peripheral blood mononuclear cells (PBMCs) and polymorphonuclear leukocytes, respectively, but viral titres were generally low. Fractionation of PBMCs into monocyte- and B, T4, and T8 lymphocyte-enriched subpopulations showed no pattern in the viral loads that might suggest the preferential association of TTV to one or more specific cell types. TTV-negative PBMCs absorbed measurable amounts of virus when incubated with infected plasma at 4 degrees C. Furthermore, cultures of TTV-negative phytohaemagglutinin-stimulated PBMCs exposed in vitro to virus-positive plasma and faecal extracts released considerable levels of infectious TTV into the supernatant fluid and the same was true for TTV-positive stimulated PBMCs. These results indicate that, whereas freshly harvested resting PBMCs seem to produce little, if any TTV, stimulated PBMCs actively replicate the virus.     

Prevalence and risk factor analysis of TTV infection in prostitutes

TTV, a DNA virus, has been isolated from patients with non-A to non-E post-transfusion hepatitis. In the past it was assumed that TTV was transmitted parenterally. It is unclear whether sexual contact leads to transmission of this virus. In this study, two sets of TTV-specific polymerase chain reaction primers were used to detect serum TTV DNA in 140 prostitutes and 136 controls. The prevalence of TTV DNA in prostitutes was significantly higher than in the control group (46/140 [32.9%] vs. 29/136 [21.3%]; P = 0.043). There was no significant difference in the prevalence of positive antibody to hepatitis A virus (anti-HAV) in either group (87.8% for prostitutes, 85.3% for controls). No particular risk factor was significantly associated with positive TTV DNA in prostitutes. In summary, TTV is highly prevalent in prostitutes. Transmission of TTV via sexual contact is not as efficient as transmission of hepatitis C and D viruses and GB virus-C hepatitis G virus. The high prevalence of TTV in controls indicates that there are diverse routes of transmission of this virus.     

Real-time PCR methods for independent quantitation of TTV and TLMV

There is considerable interest in the possible clinical effects of the human circoviruses TT virus (TTV) and TTV-like mini virus (TLMV). Most people appear to have at least one of these viruses replicating actively in their bodies, thus mere correlation of the presence of virus and disease states are probably less informative than a quantitative analysis of viraemia. Real-time PCR based methods, with either SYBR Green or TaqMan probe, designed to quantitate selectively TTV and TLMV are described. The suggested TaqMan-based protocols were suitable for quantitation of viruses in the range of 10(2)-10(9) copies/ml of sample; and proved, by sequencing of PCR products, to be specific for each of the two viruses.