Localization and changes of hemorrhagic fever with renal syndrome virus in lymphocyte subpopulation

Authors used double-labeled immunocytochemistry for studying the localization and distribution of hemorrha-gic fever with renal syndrome virus (HFRSV) antigen in the peripheral blood lymphocyte subpopulations in HFRS patients and the quantitative changes of these subpopulations. It was found that HFRSV might infect dif-ferent subpopulations of lymphocytes. The infection rate of B cells (31.3%) was significantly higher than that of Tcells (8.5%), and the infection rate of CD8~+ subpopulation (10.5%) was significantly higher than that of     

Localization and changes of hemorrhagic fever with renal syndrome virus in lymphocyte subpopulation.

Double-labeled immunocytochemistry was used for studying the localization and distribution of hemorrhagic fever with renal syndrome virus (HFRSV) antigen in the peripheral blood lymphocyte subpopulations in HFRS patients and the quantitative changes of these subpopulations. It was found that HFRSV might infect different subpopulations of lymphocytes. The infection rate of B cells (31.3%) was significantly higher than that of T cells (8.5%), and the infection rate of CD8+ subpopulation (10.5%) was significantly higher than that of CD4+ subpopulation (6.1%). The infection rates of HLA-DR+ cells and Tac+ cells were 2.5% and less than 0.5%, respectively. From febrile to oliguric stage, the number of CD8+ cells increased gradually, leading to a decrease and reverse of CD4/CD8 ratio, while the number of CD4+ cells remained almost unchanged. The appearance and disappearance of HFRSV in lymphocytes were closely associated with the dynamic changes of CD4/CD8 ratio and HLA-DR and Tac expressions. The results suggested that there were certain differences in HFRSV infectivity of different lymphocyte subpopulations, which might be due to the disorders of immunoregulation and immunopathology of HFRS.

     

In situ hybridization detection of persistent infection and viral RNA distribution in laboratory rats with hemorrhagic fever

ＩｎｓｉｔｕｈｙｂｒｉｄｉｚａｔｉｏｎｄｅｔｅｃｔｉｏｎｏｆｐｅｒｓｉｓｔｅｎｔｉｎｆｅｃｔｉｏｎａｎｄｖｉｒａｌＲＮＡｄｉｓｔｒｉｂｕｔｉｏｎｉｎｌａｂｏｒａｔｏｒｙｒａｔｓｗｉｔｈｈｅｍｏｒｒｈａｇｉｃｆｅｖｅｒｗｉｔｈｒｅｎａｌｓｙｎｄｒｏｍｅ...     

Detection of c-myc gene expression in nasopharyngeal carcinoma by nonradioactive in situ hybridization and immunohistochemistry.

Ｄｅｔｅｃｔｉｏｎｏｆｃ┐ｍｙｃｇｅｎｅｅｘｐｒｅｓｉｏｎｉｎｎａｓｏｐｈａｒｙｎｇｅａｌｃａｒｃｉｎｏｍａｂｙｎｏｎｒａｄｉｏａｃｔｉｖｅｉｎｓｉｔｕｈｙｂｒｉｄｉｚａｔｉｏｎａｎｄｉｍｍｕｎｏｈｉｓｔｏｃｈｅｍｉｓｔｒｙＬｕｏＪｕｎｌｉ罗均利，Ｘ...     

The distribution and duration of hantaan virus in the body fluids of patients with hcmorrhagic fever with renal syndrome

The distribution and duration of Hantaan virus (HTNV) in body fluids of patients were studied byimmunofluorescence,reverse passive hemagglutination,and cell culture assays. Virus antigen of hemorrhagic feverwith renal syndrome in peripheral blood mononuclear cells (PBMCs) was usually present before day 11 of the dis-ease ,especially from days 4--7,Virus isolates were more rcadily recovered from plasma early in the course of the     

Effects of Hantaan virus on human endothelial cells and their significance in pathogenesis of hemorrhagic fever with renal syndrome.

The effects of Hantaan virus (HTNV) on human endothelial cells (HECs) were investigated both in vivo and in vitro. The 76-118 strain or SR-11 strain of HTNV were inoculated into HECs monolayers respectively, and the virus antigens could be detected on the seventh day of the first passage after inoculation by immunofluorescent technique. The HTNV could also be isolated through cultures of Vero E-6 cells. HTNV particles and inclusion bodies together with various changes in the organelles were observed in the infected cells by transmission electronic microscopy (TEM), and the immunoenzyme positive virus particles were seen by immunoelectronic microscopy. Samples of skin biopsy were individually obtained from 14 cases of hemorrhagic fever with renal syndrome on the second to the fifth day after the onset of the illness. It was found that HTNV antigens were widely distributed in the cytoplasm of endothelial cells of the samples from 5 out of 14 cases by avidin-biotin-peroxidase complex staining, and morphologic changes of the endothelial cells similar to those observed in vitro, were also seen by TEM. The results indicated that HEC is one of the target cells susceptible to HTNV. The virus could invade and propagate in HECs, and could induce damage to the latter.

     

Hemorrhagic fever with renal syndrome. Separation of human peripheral blood T and B cells and detection of viral antigen.

In this study, total blood lymphocytes were prepared from patients with hemorrhagic fever with renal syndrome (HFRS) by a density gradient on Ficoll-Hypaque. T and B cells were then purified by passing the total lymphocytes over a nylon wool column. The purities of total lymphocytes, T cells and B cells were 97.8 +/- 2.3%, 91.6 +/- 4.5% and 74.2 +/- 12.1%, respectively. Also, after modification of cell fixation and smear drying, the number of cells were increased and the time needed for slide preparation was shortened. Detection of viral antigen by immunofluorescence assay using monoclonal antibodies to Hantavirus (HTNV) showed that the total lymphocytes. T cells and B cells were infected by HTNV during the early stages of HFRS and no specific fluorescence was seen in the cells from the late diuretic phase to convalescent phase. The results suggest that virus replication in blood lymphocytes may partly contribute in the early stages to the impairment of cell immune response and in vivo spread of HTNV to its target sites.

     

Epidemiologic surveillance on the hemorrhagic fever with renal syndrome in China.

This article summarizes the contents and results of extensive surveillance on hemorrhagic fever with renal syndrome (HFRS) in China. Data from surveillance on human HFRS, including morbidity and case fatality rate, trend of epidemics, periodicity of epidemics, factors related to epidemics, types of epidemic areas, environment structure of natural nidi, geographic distribution, seasonal distribution, age distribution, sex distribution and occupation distribution, as well as surveillance on animal infection with HFRS virus were collected and analysed.

     

Characteristics of immunocomplex in autopsy tissues of hemorrhagic fever with renal syndrome.

Using repeated PAP or repeated PAP and ABC immunocytochemical methods, we were able to demonstrate viral antigens, Ig and CIq in the tissues of 20 autopsy materials that had been preserved for 3-30 years. Serial paraffin sections were stained with the first antibodies against both viral antigens (G2 and Np) and human IgG, IgM, C3 and Clq. Immunocomplex, composed of viral antigen, IgG and Clq were found diffusely in the cells of each organ and extracellalarly in the sera, various secretions and exudates. When stained by A25 etc, coarse granular antigen or inclusion bodies were found without demonstrable Ig and Clq. It was concluded that the immunocomplex in tissues of epidemic hemorrhagic fever in Shaanxi Province, China was both intracellular and extracellular and was perhaps soluble due to antigens in excess, with characteristics quite different from that of other immune diseases.     

肝脏中肾综合征出血热病毒感染与c－myc基因的表达以及免疫球蛋白的异常分布

目的：观察比较人和动物肝脏感染肾综合征出血热（ＨＦＲＳ）病毒后病变的异同．方法：以原位分子杂交和免疫组化检测ＨＦＲＳ尸检及自然感染大鼠肝脏中病毒ＲＮＡ，病毒抗原及免疫球蛋白（Ｉｇ）的定位和分布及尸检肝脏中ｃ－ｍｙｃ基因．结果：尸检肝脏中病毒ＲＮＡ，病毒抗原和Ｉｇ的阳性率分别为８４．２１％（１６／１９），８８．２３％（１５／１７）和５３．３０％（８／１４），大鼠肝脏阳性率分别为９２．９０％（７９／８５），９３．１０％（２７／２９）和５４．５０％（１８／２９）；尸检肝脏ｃ－ｍｙｃ基因阳性率为５０．００％（８／１６）；尸检阳性者主要为死于休克期和重型病例；类似于人尸检肝脏中的病变未在大鼠肝脏中观察到．结论：与其它脏器相比，肝脏具有较高的病毒感染率．肝细胞异常Ｉｇ阳性可能是由病毒感染造成的肝细胞的可逆性损伤的体现，而肝细胞的坏死可能是损伤严重者．肝脏局部血液循环障碍是造成大片坏死的主要因素．Ｃ－ｍｙｃ基因的表达可能是肝脏抗损伤的一种反应．     

原位分子杂交检测大鼠肾综合征出血热病毒隐性感染和病毒RNA的体内分布

目的和方法：用异羟基洋地黄毒苷配基－１１－ｄＵＴＰ随机引物延伸法分别标记肾综合征出血热（ＨＦＲＳ）汉坦株病毒Ｍ与ＳＲＮＡ全片段ｃＤＮＡ作为探针，用核酸原位分子杂交法检测某动物所一批大鼠ＨＦＲＳ病毒的隐性感染及病毒ＲＮＡ的组织细胞定位和分布．结果：９８只大鼠中８８只病毒ＲＮＡ阳性，阳性率８９．７９％，阳性率最高的脏器是肝脏７９／８５，其次为心脏２２／８３，再次为肾脏１４／７９，其余脏器阳性率较低．阳性部位主要在各脏器的实质细胞胞浆内，呈细颗粒状，未见于核内．结论：结果说明本批大鼠ＨＦＲＳ病毒隐性感染率极高；大鼠ＨＦＲＳ病毒ＲＮＡ的阳性检出率明显高于同批大鼠病毒抗原和血清抗体的阳性检出率，说明原位分子杂交检测病毒ＲＮＡ对确定动物病毒隐性感染具有重要的流行病学意义．病毒ＲＮＡ主要存在于肝脏，提示肝脏可能是病毒的主要潜伏部位，病毒可能主要由粪便排除．     

原位分子杂交检测肾综合征出血热尸检肝组织中c－myc基因表达

目的和方法：作者用异羟基洋地黄毒苷配基－１１－ｄＵＴＰ随机引物延伸法标记ｃ－ｍｙｃｃＤＮＡ为探针，用核酸原位分子杂交法检测了１６例西安地区肾综合征出血热（ＨＦＲＳ）尸检肝组织内ｃ－ｍｙｃ基因表达．结果：８例尸检肝组织中，ｃ－ｍｙｃ有不同程度的表达，阳性检出率为５０％．杂交阳性信号呈颗粒状主要出现在肝细胞胞浆内，在发病后６～８ｄ死亡的病例肝组织ｃ－ｍｙｃ基因表达强而弥漫，仅１例见于核内呈散在分布．结论：肝细胞中ｃ－ｍｙｃ基因的表达可能主要系ＨＦＲＳ时肝脏抗损伤和肝细胞增殖的一种表现，另外还可能参与ＨＦＲＳ肝组织内常见的肝细胞凋零现象．     

肾综合症出血热肾组织中病毒感染及病毒抗原的来源和意义

目的：明确肾综合征出血热（ＨＦＲＳ）肾组织中病毒感染及抗原的来源和意义。方法：应用免疫组化和原位分子杂交技术，分别对西安、沈阳、上海、江西和广州地区收集的３０例ＨＦＲＳ尸检肾组织进行了研究；结果：２８例进行病毒抗原检测，有２６例阳性（２６／２８／３０）；２８例进行病毒ＲＮＡ检测，２６例阳性（２６／２８／３０）。西安及沈阳病例，病毒抗原和ＲＮＡ阳性部位主要是肾小管上皮细胞。上海及江西病例，病毒抗原定位于肾间质血管内皮，血管壁和肾小管上皮；而病毒ＲＮＡ主要定位于肾间质血管和血管内皮，少数病例伴肾小管上皮细胞阳性；１例上海病例肾小管上皮细胞胞核病毒ＲＮＡ阳性。广州病例，病毒抗原主要定位于肾血管平滑肌细胞；在３例西安病例和６例上海病例肾脏远曲小管和集合管上皮细胞中检测到ＮＰ和ＨＡ抗原阳性的胞浆内病毒包涵体（ＣＩＢ）。在病变严重的肾小管和间质血管部位可同时检测到ＩｇＧ，ＩｇＭ和Ｃ３阳性；少数病例肾小球也有不连续阳性，但较前两者弱。结论：ＨＦＲＳ病毒可感染肾脏中肾小管上皮和血管内皮，并在肾小管上皮细胞产生包涵体，不同地区病例肾脏中病毒感染的部位不同，可能与不同地区病例所感染的病毒毒株或血清型不同有关。病毒感染所诱发的Ⅱ     

肾综合征出血热并发意识障碍33例分析

目的 探讨肾综合征出血热 (HFRS) 并发意识障碍的病因及影响预后的相关因素.方法 33例有意识障碍的HFRS患者按预后分为两组,其中死亡组16人;治愈组17人,回顾性分析其病因、症状、体征和实验室检查结果.结果 意识障碍的病因中脑出血占21% (病死率71%) ,脑水肿、脑疝占24% (病死率100%) .死亡组病例休克和少尿的持续时间比治愈组延长,低钠血症和腔道出血的发生率高于治愈组 (P<0.01和P<0.05) .死亡组血小板计数明显下降.血肌酐和血清丙氨酸氨基转移酶 (ALT) 显著升高,与治愈组比较差异均有显著性.结论 肾综合征出血热发生脑出血和脑疝预后不良.休克期和少尿期延长、腔道出血、血小板减少和低钠血症可能是影响肾综合征出血热并发意识障碍患者预后的重要因素,应引起重视.     

肾综合征出血热发病机制和早期诊断现状

肾综合征出血热（Hemorrhagic fever with renal syndrome,HFRS）是由啮齿类动物携带的汉坦病毒（Hantaan virus,HV）而传播的一种急性传染病。该病流行较广泛,危害较严重,已成为一个全球性的公共卫生问题。其发病机制较复杂,早期诊断极为关键,本文对HFRS的发病机制、早期诊断的几个主要方面作扼要的阐述。     

肾综合征出血热患者的细胞免疫功能研究

目的：研究肾综合征出血热患的细胞免疫功能异常情况。方法：应用流式细胞仪检测CD4^ T淋巴细胞、CD8^ T淋巴细胞。结果肾综合征出血热患病程早期，后期与正常人比较，均有CD4^ T淋巴细胞计数降低（p＝0．000，p=0.000),CD8^ T淋巴细胞计数无明显升高（p=0.000,p=0.002)，CD4^ ／CD8^ 明显降低（p=0.000,p=0.000)。CD4^ T淋巴细胞计数病程早期与病程后期无明显差异（p=0.985)，CD8^ T淋巴细胞计数病程早期明显高于病程后期（p=0.000)，CD4^ ／CD8^ 病程早期明显低于病程后期（p=0.000).结论：肾综合征出血热患发病时存在明显细胞免疫功能异常，在病程后期逐渐恢复。     

74例肾综合征出血热的流行病学及临床特征分析

目的 分析肾综合征出血热（hemorrhagic fever with renal syndrome，HFRS）的流行病学及临床特征，为HFRS的早期临床诊断提供参考依据。方法 回顾性分析大连医科大学附属第二医院2005年1月至2018年3月收治的实验室确诊为HFRS的患者74例，其中男48例，女26例，年龄14~86岁，分析其临床症状和体征、实验室检查、误诊情况、治疗费用和转归情况，应用SPSS 25.0对数据进行统计分析。结果 临床症状中发热症状最多见，占93.24%，其次为乏力、头痛、腰痛、恶心、呕吐、少尿或无尿，具有典型"三红三痛"临床表现病例仅21例，占28.37%。临床分型以轻型和中型居多。实验室检查以肾功能损伤、急性血小板减少、肝功能损伤为主。首诊误诊病例共19例，误诊率25.7%。结论 HFRS具有典型临床表现者少，误诊率高。临床医生需加强对该病的认识。