艾滋病患者的肝脏病理学研究

目的 观察艾滋病(AIDS)患者肝组织的病理改变。方法 对11例无症状人类免疫缺陷病毒 (HIV)感染者和3例 AIDS 患者肝活体组织检查(肝活检)以及12例尸体解剖肝脏进行组织学观察，采用 免疫组织化学对肝活检组织及尸体解剖肝组织中的HIV-1外膜蛋白 gp120和衣壳蛋门 p24抗原进行检测。结 果 14例肝活检组织中检测出巨细胞病毒感染1例，11例肝脏库普弗细胞和部分淋巴细胞中有 HIV-1外 膜蛋白 gp120或(和)衣壳蛋白 p24抗原表达，肝细胞呈阴性。尸体解剖肝组织中检出巨细胞病毒感染5例， 分枝杆菌及弓形虫感染各3例，HIV 1衣壳蛋白 p24抗原阳性5例。结论 AIDS 患者肝组织中存在 HIV 1 感染，活检肝组织病例中机会性感染的几率较低。在尸体解剖肝组织中，机会性感染较为多见，并常为全身 机会性感染的局部表现。     

179例HIV／AIDS病人肝脏损害的临床研究

目的探讨艾滋病病毒（HIV）感染者／艾滋病（AIDS）病人的肝脏功能损害情况及其影响因素。方法回顾分析179例HIV／AIDS病人临床资料,根据肝脏功能损害程度,分为肝功能正常组、肝功能轻度损害组、中度损害组、重度损害组,比较各组间相关情况。结果肝功能损害组使用肝毒性药物、合并HBV和／或HCV感染的比例均高于肝功能正常组,两者比较差异有显著的统计学意义（P值分别为0．039、0．030、0．003）。中重度肝功能损害组使用肝毒性药物的比例高于轻度损害组,两者比较差异有显著的统计学意义（P=0．037）。结论HIv／AIDS病人肝脏损害发生率高,而男性、使用肝损害药物、合并HBV和／或HCV是HIV／AIDS病人发生肝脏损害的危险因素。     

AIDS患者胃黏膜病理改变与HIV感染的关系

目的探讨AIDS患者胃黏膜病理改变与HIV感染的关系。方法选取AIDS期HIV慢性感染患者25例,PCR法制备地高辛标记HIV-1-LTR、gag基因的双链cDNA探针,对胃黏膜组织切片采取核酸原位杂交方法观察HIV感染情况和黏膜病理改变。结果AIDS患者胃黏膜主要表现为慢性浅表性胃炎（CSG）和慢性萎缩性胃炎（CAG）。且AIDS患者胃黏膜组织多种细胞中均有HIV阳性显色：淋巴滤泡细胞、固有层中散在的单个核细胞（似淋巴细胞和巨噬细胞）、胃黏膜上皮细胞、腺上皮细胞、极少数间质细胞。CSG组与CAG组胃黏膜HIV的阳性表达率无明显差异（P〉0．05）。结论HIV／AIDS疾病的进程与胃黏膜局部病理改变之间可能存在一定关联性,但HIV感染可能并非发挥重要作用。     

Panleucogating方法计数CD4^＋T淋巴细胞的研究进展

摘要：目的阐述Panleucogating方法计数艾滋病病毒（HIV）感染者／AIDS病人（HIV／AIDS病人）CD4^＋T淋巴细胞（CO4细胞）的方法、发展和应用。方法收集HIV／AIDs病人CD4^＋细胞检测方法的文献,分析Panleucogating方法的可行性及其优势和不足。结果发现Panleucogating方法与常规方法检测的CD4^＋细胞数值之间存在较高的相关性。该方法具有检测成本低、操作简单、检测迅速等优势。结论Panleucogating方法可用于HIV／AIDS病人CD4细胞计数的检测,在经济欠发达地区有应用价值。     

HIV/AIDS病人泌尿生殖道组织HIV-1总DNA的检测与分析

目的探讨泌尿生殖道组织作为艾滋病病毒(HIV)感染者/艾滋病(AIDS)病人(简称HIV/AIDS病人)HIV储存库的可能。方法 20例接受高效抗反转录病毒治疗(HAART)至少6个月,需要行包皮切除术或泌尿道/生殖道腔镜手术的HIV/AIDS病人,检测标本组织和外周血单个核细胞(PBMC)的HIV-1总脱氧核糖核酸(DNA)。结果 20例病人平均年龄(45.8±14.7)岁,接受HAART(6～36个月),平均(14.1±7.9)个月,外周血HIV-1核糖核酸(RNA)均低于检测下限20拷贝/mL,CD4~+T淋巴细胞(简称CD4细胞)计数平均(417.3±151.3)个/μL,CD4细胞/CD8~+T淋巴细胞(简称CD8细胞)(CD4/CD8细胞)比值平均(0.55±0.35),泌尿生殖道组织和PBMC中均能检测到HIV-1总DNA,载量分别为(85.9～89 300)拷贝/10~6个细胞(中位数1 034.12拷贝/10~6个细胞)及(6.83～321 000)拷贝/10~6个细胞(中位数550拷贝/10~6个细胞),两者呈正相关(r=0.53,P=0.018)。但泌尿生殖道组织HIV-1总DNA载量与CD4细胞计数及CD4/CD8细胞比值变化均无明显相关性(r=-0.002,P=0.99;r=0.12,P=0.61)。结论泌尿生殖道组织可能是HIV感染者的病毒储存库之一。     

人类免疫缺陷病毒感染者重叠输血传播病毒感染的临床研究

目的 了解中国地区人类免疫缺陷病毒（HIV-1）感染者重叠输血传播病毒（TTV）感染的流行情况，并探讨重叠感染TTV对CD4^＋T淋巴细胞计数及HIVRNA病毒载量的影响。方法收集55例HIV-1感染者的血浆标本，采用荧光定量PCR法检测TTVDNA，流式细胞法检测T淋巴细胞亚群，bDNA方法检测HIVRNA。结果HIV感染者中TTVDNA检出率及TTVDNA滴度明显高于健康对照组（P〈0．05）。HIV／TTV重叠感染者CD4^＋T淋巴细胞计数低于单纯HIV感染者。HIVRNA病毒载量高于单纯HIV感染者，两组比较差异有统计学意义（P〈0．05）。结论中国地区HIV感染者易重叠感染TTV，且滴度较高。重叠感染TTV可能导致细胞免疫功能进一步下降，加快病毒复制，促进HIV病程的进展。     

中国人类免疫缺陷病毒-1感染者细胞毒性T淋巴细胞特异性应答

目的 探讨中国人类免疫缺陷病毒（HIV）／艾滋病（AIDS）患者HIV特异性细胞毒性T淋巴细胞（CTL）应答的特征。方法 以HIV-1 B、C亚型构建的2个全基因组肽库作为抗原，通过酶联免疫斑点（ELISPOT）法检测HIV／AIDS患者HIV特异性CTL应答。结果 无论HIV-1 B亚型还是HIV-1 C亚型所构建肽库的应答效应和频率主要集中在Gag和Nef蛋白，其他蛋白也有不同程度的应答。HIV-1 B、C亚型间应答比较，整体范围大致相同，但单个肽段水平还存在着一定差异，B亚型应答频率最高的是Nef的GPKEPFRDYVDRFYKTLR（5／17，29．4％）和Gag的LWVYHTQGYFPDWQNY（5/17，29．4％），C亚型应答频率最高的是Gag的GPKEPFRDYVDRFFKTLR应答频率为35．29％。结论 中国人群CTL应答多集中在Gag和Nef蛋白，B、C亚型在单个肽段水平略有差异。提示中国人群的CTL应答研究对设计针对中国人群的HIV疫苗有较重要的意义。     

区分HIV-1p24抗原和HIV抗体检测方法的临床应用

目的了解艾滋病病毒(HIV)-1p24抗原和抗体联合检测在临床应用的价值。方法总结从2012-2016年HIV-1p24抗原和抗体联合检测并且能区分是抗原还是抗体阳性的临床结果,并和不能区分抗原抗体的四代酶联试剂结果比较,对其中HIV-1p24抗原阳性的样本进行CD4+T淋巴细胞及HIV核糖核酸(RNA)的检测。结果 2012-2016年共检测HIV-1p24抗原和抗体联合检测样本10 476例,其中HIV抗体阳性777例,HIV-1p24抗原阳性36例,其中抗原抗体同时阳性8例,单独HIV-1p24抗原阳性28例。结论 HIV-1p24抗原和抗体联合检测敏感性高、特异性强,尤其是能将抗原和抗体阳性区分开来,大大缩短了窗口期,提高了HIV急性期感染的检出率。     

HIV/AIDS病人HAART致肝功能损害66例分析

目的探讨高效抗反转录病毒治疗（HAART）对艾滋病病毒（HIV）感染者/艾滋病（AIDS）病人（简称HIV/AIDS病人）肝功能的影响。方法回顾性分析HIV/AIDS病人抗病毒治疗24个月内肝功能的变化情况。结果共计调查755例HIV/AIDS病人,肝功能损害发生率为8.7%（66/755）,以单项转氨酶或胆红素升高为主,轻、中度肝损害占84.8%（56/66）,发生异常的时间为30-485天,中位数75天。其中含奈韦拉平（NVP）方案治疗者肝功能损害的发生率为7.1%（33/467）、含依非韦伦（EFV）方案发生率为11.5%（33/288）。肝损害级别：1级34例,2级22例,3级10例。结论 HIV/AIDS病人在抗病毒治疗过程中轻中度肝损害常见,应严密观察,及时处理,以保证HAART的顺利进行。     

HGV/GBV-C与HCV混合感染者肝组织中相关病毒表达的免疫组化研究

目的 了解HGV/GBV-C与HCV混合感染者肝组织HGV/GBV-C相关抗原的分布状况,探讨HGV/GBV-C对肝脏的损害机制。方法 以抗HGV/GBV-C NS5单克隆抗体或抗HCV NS3单克隆抗体为试剂,采用免疫组织化学方法检测肝炎病人肝组织中HGV/GBV-C、HCV相关抗原表达。结果 56例肝炎病肝组织中HGV/GBV-C相关抗原表达阳性率为26.79％(15/56);HCV NS3抗原表达阳性率为39.29％(22/56)。HGV/GBV-C NS5抗原表达阳性信号主要位于肝细胞胞浆中,染色阳性细胞周围可见淋巴细胞浸润。结论 肝细胞中存在HGV/GBV-C相关抗原表达,其编码产物可能作为一种靶抗原,诱发免疫病理反应,免疫损伤可能是其发病机制之一。     

Identification and quantitation of HIV-1 in the liver of patients with AIDS.

OBJECTIVE: To detect and quantify HIV-1 in the liver in vivo. DESIGN: Fourteen liver biopsy samples and corresponding blood lymphocytes and monocytes from patients with AIDS were studied for HIV-1 using quantitative polymerase chain reaction (PCR). In addition, expression of HIV-1 antigen and messenger (m) RNA in 10 autopsy liver specimens was examined by immunohistochemistry and in situ hybridization. RESULTS: The amount of HIV-1 DNA in nine liver samples ranged from 850 to 27,000 copies per 10(6) cells, with mean and median values of 8150 and 3500 copies per 10(6) cells, respectively. Five other samples had no detectable HIV-1 DNA by PCR. Intracellular expression of HIV-1 antigen and mRNA was also detected in both Kupffer cells and hepatocytes by in situ studies. CONCLUSION: These findings strongly indicate that HIV-1 could replicate in the liver of a majority of patients with AIDS.     

High frequency of chimerism in transplanted livers

Recent studies have shown that primitive stem cells can mobilize and differentiate into hepatocytes. We investigated the time and extent in which cells of recipient origins could differentiate into hepatocytes and other cells in human liver allografts. Microsatellite analysis, which can assess quantitatively the proportions of recipient and donor DNA, was performed in posttransplantation liver biopsy specimens from 17 patients at various times. Combined fluorescence in situ hybridization (FISH) for Y chromosome and immunofluorescence for different cell types was also performed in 10 of these cases with sex mismatch. Organ chimerism in the transplanted livers was found to be of variable extent, and the recipients' DNA in the posttransplantation liver biopsy specimens (excluding portal tracts) amounted up to 50%. The recipient DNA in the posttransplantation liver biopsy specimens increased after liver transplantation by as early as 1 week, peaked at around 30 to 40 weeks, and could be shown 63 weeks after transplantation. Most (64%-75%) of the recipient-derived cells showed macrophage/Kupffer cell differentiation. Only up to 1.6% of the recipient-derived cells in the liver grafts showed hepatocytic differentiation in the liver grafts and made up 0.62% of all hepatocytes of both donor and recipient origins. These livers had mild or minimal injury histologically. In conclusion, our results show that most of the recipient-derived cells in the liver allografts were macrophages/Kupffer cells and only a small proportion of hepatocytes was recipient derived. However, with regard to recipient-derived hepatocytes, our data cannot distinguish between transdifferentiation and cell fusion.     

Heat-denatured human immunodeficiency virus type 1 protein 24 antigen: prognostic value in adults with early-stage disease

CD4(+) lymphocyte count and human immunodeficiency virus (HIV) type 1 RNA level are useful for determining when to initiate antiretroviral therapy but are not used widely in developing countries due to the high cost. Heat-denatured protein 24 (p24) antigen is an inexpensive assay that predicts disease progression among persons with advanced disease but has not been assessed among persons with early-stage disease. Plasma levels of heat-denatured p24 antigen were quantified in baseline study-visit specimens obtained from injection drug users enrolled in a longitudinal cohort study of HIV-1 infection. Of the 494 study participants (median initial CD4(+) lymphocyte count, 518 lymphocytes/mm(3)), 90 (18%) progressed to acquired immunodeficiency syndrome within 5 years. p24 antigen level correlated with both CD4(+) lymphocyte count (r=-0.34; P<.0001) and HIV-1 RNA level (r=0.55; P<.0001). p24 antigen level >5 pg/mL predicted disease progression, comparable with that of cutoff CD4(+) lymphocyte count <350 lymphocytes/mm(3) and HIV-1 RNA level >30,000 copies/mL. Heat-denatured p24 antigen level predicted subsequent clinical disease progression in early-stage HIV-1 infection and correlated with both CD4(+) lymphocyte count and HIV-1 RNA level.     

Localization of dengue virus in naturally infected human tissues, by immunohistochemistry and in situ hybridization

Dengue viral antigens have been demonstrated in several types of naturally infected human tissues, but little is known of whether these same tissues have detectable viral RNA. We studied tissue specimens from patients with serologically or virologically confirmed dengue infections by immunohistochemistry (IHC) and in situ hybridization (ISH), to localize viral antigen and RNA, respectively. IHC was performed on specimens obtained from 5 autopsies and 24 biopsies and on 20 blood-clot samples. For ISH, antisense riboprobes to the dengue E gene were applied to tissue specimens in which IHC was positive. Viral antigens were demonstrated in Kupffer and sinusoidal endothelial cells of the liver; macrophages, multinucleated cells, and reactive lymphoid cells in the spleen; macrophages and vascular endothelium in the lung; kidney tubules; and monocytes and lymphocytes in blood-clot samples. Positive-strand viral RNA was detected in the same IHC-positive cells found in the spleen and blood-clot samples. The strong, positive ISH signal in these cells indicated a high copy number of viral RNA, suggesting replication.     

Rapid activation of lymph nodes and mononuclear cell HIV expression upon interrupting highly active antiretroviral therapy in patients after prolonged viral suppression

OBJECTIVE: To compare the architecture and HIV-1 RNA and Gag p24 protein expression in lymph nodes (LN) excised from individuals during chronic highly active antiretroviral therapy (HAART) with LN removed from the same patient after plasma virus rebound following the interruption of HAART. MATERIALS AND METHODS: Six HIV-1-infected patients on HAART, with CD4 cell counts greater than 350 cells/microl, and plasma HIV-1 RNA less than 50 copies/ml, underwent inguinal LN excision upon discontinuation of HAART, and again after rebound of plasma virus. Lymph nodes were evaluated by immunohistochemical staining for Gag p24 antigen and Ki67, in-situ hybridization for HIV-1 RNA and H3-histone, and transmission electron microscopy (TEM). RESULTS: LN at baseline were quiescent to mildly hyperplastic and generally contained more primary than secondary follicles. Only one LN had detectable follicular dendritic cell (FDC)-associated p24 antigen, none had HIV RNA. Few mononuclear cells (MNC) expressed RNA or p24 antigen. Plasma virus at the second biopsy ranged from 329 to 3.2 x 10(6) copies/ml. CD4 cell count decline ranged from 5 to 51% during drug hiatus, and was greatest in patients with highest viral rebound. Four of six of the second LN were more hyperplastic than the initial LN, two showed paracortical hyperplasia. MNC expression of HIV RNA in the second LN paralleled the level of plasma viremia. Increased Ki67 and H3-histone signal occurred in the second LN. CONCLUSION: Quiescent LN from individuals on HAART rapidly become hyperplastic and activated within 1-2 months after treatment interruption. As in acute HIV infection, virus expression by LN MNC parallels the rebound in plasma viremia and fall in CD4 cell count.     

Detection of HIV-1 RNA in the lamina propria of patients with AIDS and gastrointestinal disease

Thirty formaldehyde-fixed, paraffin-embedded endoscopic biopsy specimens from the colon and rectum of 25 patients with acquired immunodeficiency syndrome were examined using a [35S]HIV-RNA in situ hybridization procedure. Nine of the specimens contained cells that bound significant amounts of probe. Cells were considered positive if more than 50 grains of silver (over background) per 200 micron 2 were seen over cells that did not stain with eosin. Most of the positive cells resembled macrophages, although cells with condensed nuclei resembling lymphocytes were found. No epithelial cells expressing viral RNA were detected. Formaldehyde-fixed eosinophils gave spurious signals that could be reduced with sulfhydryl modifying agents. HIV-1 may be disseminated in the lamina propria of the gut at low concentrations in some patients but may not be detectable in others. The lower gut lining may be both a portal of initial infection with HIV and a target of disseminated HIV infection.     

Analysis of human immunodeficiency virus-infected tissues by amplification and in situ hybridization reveals latent and permissive infections at single-cell resolution.

Latent and productive viral infections are at the extremes of the spectrum of virus-cell interactions that are thought to play a major role in the ability of such important human pathogens as human immunodeficiency virus (HIV) to elude host defenses and cause disease. The recent development of PCR-based methods to amplify target sequences in individual cells in routinely fixed tissues affords opportunities to directly examine the subtle and covert virus-cell relationships at the latent end of the spectrum that are inaccessible to analysis by conventional in situ hybridization techniques. We have now used PCR in situ with in situ hybridization to document latent and permissive HIV infection in routinely fixed and paraffin-embedded tissue. In one of the first specimens we examined, a tumor biopsy from an HIV-infected individual, we found many of the lymphocytes and lymphocytes infiltrating the tumor had HIV DNA that was detectable only by PCR in situ. The fraction of positive cells varied regionally, but there were foci where most of the cells contained HIV DNA. Most of these lymphocytes and macrophages are latently infected, as we could detect HIV RNA in fewer than one in a thousand of these cells. We also detected HIV RNA, surprisingly, in 6% of the tumor cells, where the number of copies of viral RNA per cell was equivalent to productively infected cell lines. The alternative states of HIV-gene expression and high local concentration of latently infected lymphocytes and monocytes revealed by these studies conceptually supports models of lentiviral pathogenesis that attribute persistence to the reservoir of latently infected cells and disease to the consequences of viral-gene expression in this population. The magnitude of infection of lymphocytes documented in this report is also consistent with the emerging view that HIV infection per se could contribute substantially to depletion of immune cells in AIDS.     

Kupffer cell iron overload induces intercellular adhesion molecule-1 expression on hepatocytes in genetic hemochromatosis

The mechanisms underlying iron-induced liver fibrogenesis in patients with genetic hemochromatosis are poorly understood. We studied signs of Kupffer cell activation and inflammatory responses in liver biopsy specimens obtained from 15 patients with untreated and six patients with treated hemochromatosis. Immunohistochemistry was performed on 11 of the untreated and all treated patients. Three of the untreated patients (20%) had cirrhosis and eight (53%) had fibrosis. None had chronic active hepatitis (CAH). Immunohistochemistry indicated that 55% of the untreated patients had sparse intercellular adhesion molecule-1 (ICAM-1) expression by hepatocytes, and all of these had Kupffer cell iron overload. No ICAM-1 expression was seen by hepatocytes in treated patients or healthy controls. ICAM-1 was strongly expressed by hepatocytes from control patients with inflammatory liver disease. HLA-DR reactivity was seen on sinusoidal cells in all groups, but not on hepatocytes except for two of the control patients with CAH. Twenty-seven percent of the untreated hemochromatosis patients displayed moderate infiltration by CD3-positive lymphocytes. Electron microscopy of samples from untreated hemochromatosis patients showed hypertrophic Kupffer cells containing iron-rich remnants of phagocytosed hepatocytes. Fat-storing cells close to iron-laden hepatocytes contained multiple lipid droplets and adjacent collagen fibril bundles. Thus, in patients with untreated genetic hemochromatosis and Kupffer cell iron overload, hepatocytes occasionally express ICAM-1. In regions with heavy iron overload, Kupffer cell hypertrophy and transition of fat-storing cells are seen. Our findings indicate that release of factors from iron-loaded, activated Kupffer cells is of importance for the transformation of fat-storing cells and increased collagen deposition seen in genetic hemochromatosis.