No association of borna disease virus with psychiatric disorders among patients in northern Kyushu, Japan

There is controversy over the prevalence of Borna disease virus (BDV) antibodies and its RNA in the peripheral blood mononuclear cells (PBMCs) of psychiatric patients, and the contribution of BDV to human psychiatric disorders. We examined 299 plasma and 229 PBMC samples. No plasma samples were positive for BDV-p40, p24 or gp18 antibodies by western blot analysis. The prevalence of BDV RNA in the psychiatric (schizophrenic) patients (1.8%) was not significantly different from that in the healthy volunteers (0.6%). The nucleotide sequences of BDV p40 and p24 were highly conserved with those of BDV He/80. Our results suggested that there is a lack of association between BDV infection and psychiatric disorders among the patients in Northern Kyushu, Japan.     

Immunological and PCR analyses for Borna disease virus in psychiatric patients and blood donors in Japan

The involvement of Borna disease virus (BDV) in psychiatric diseases in humans remains controversial. T-cell memory response and seroprevalence of BDV in patients with psychiatric disorders and blood donors in Japan were evaluated collectively by Western blot (WB) analysis with inhibition test, electrochemiluminescence immunoassay, immunofluorescence assay, and T-cell proliferative response as well as detection of BDV p24 RNA in peripheral blood mononuclear cells (PBMCs). Positive proliferative responses to both BDV p40 and p24 proteins were detected in 9% of patients with mood disorders (4 of 45), 4% of schizophrenic patients (2 of 45), and 2% of blood donors (1 of 45). By WB analysis, the antibody to BDV p40 was detected only in 2% of patients with mood disorders (1 of 45). The BDV p24 antibody was detected in 2% of patients with mood disorders (1 of 45) and 9% of schizophrenic patients. (4 of 45) No plasma reacted with both BDV proteins. The finding of a lower seroprevalence than previously reported suggests the presence of false-positive cases in the previous report. BDV RNA was detected only in 2% of patients with mood disorders (1 of 45). In these three serological assays, T-cell responses, and PCR analysis, there was no significant difference in the prevalence among the three groups. However, we found three psychiatric patients who were positive for both BDV antibodies and T-cell proliferative responses and one patient who was positive for BDV RNA in PBMCs. These findings suggest the usefulness of the proliferative T-cell response and that certain individuals are infected with BDV or a BDV-related virus.     

Demonstration of Borna disease virus (BDV) in specific regions of the brain from horses positive for serum antibodies to BDV but negative for BDV RNA in the blood and internal organs

Sero- and molecular-epidemiological studies on Borna disease virus (BDV) infection show that BDV RNA is not always detected in the peripheral blood mononuclear cells (PBMCs) from serum anti-BDV antibody-positive individuals such as horses, sheep, cattle, cats, and humans. In this study we demonstrated BDV RNA signals by polymerase chain reaction only in restricted regions of the brain from horses with locomotor disease. Four of six horses examined showed apparently positive reactions for anti-BDV antibodies. Specific regions of the brain of these four horses were positive for BDV RNA but the internal organs, lymph nodes, and PBMCs were negative. Histological studies of their brains revealed no apparent histological abnormalities such as inflammatory reactions. These results suggest that BDV chronically infects certain restricted regions of brain in seropositive horses. Received: 6 January 1997     

新疆伊犁地区马和驴博尔纳病病毒自然感染的调查

目的 调查新疆伊犁地区伊犁马和伊犁驴博尔纳病病毒(Boma disease virus,BDV)流行现状,分析BDV种系来源.方法 采用荧光定量巢式实时逆转录聚合酶链反应(fluorescence quan-titative nested RT-PCR,FQ-nRT-PCR),对新疆伊犁地区518匹伊犁马和206头伊犁驴外周血单个核细胞 (peripheral blood mononuclear cell,PBMC)进行BDV p24基因片段检测.对榆测阳性结果 的标本通过BDV p40和质粒标准品的检测进行进一步验证并测序,对其进行基因同源性、氨基酸序列和系统发生分析.结果 5例伊犁马和4例伊犁驴血标本BDV p24榆测阳性,阳性率分别为0.97%和1.94%;9例标本BDV tn0片段检测均阳性,质粒标准品榆测均阴性;BDV p24扩增产物序列与He/80株的同源性为100%.结论 新疆伊犁地区伊犁马和伊犁驴中可能存在BDV的自然感染,该地区BDV流行株与He/80株存在同源性.     

临床病毒性脑炎标本的博尔纳病病毒核酸检测

目的 探讨临床病毒性脑炎(viral encephalitis,VE)患者与博尔纳病病毒(Borna disease virus,BDV)感染的关系,分析BDV感染的病毒性脑炎患者临床特征.方法 用荧光定量巢式逆转录聚合酶链反应(FQ-nRT-PCR)方法检测病毒性脑炎患者及非感染性疾病施行蛛网膜下腔阻滞麻醉的手术患者脑脊液单个核细胞(cerebrospinal fluid mononuclear cell,CSFMC)中BDV p24基因片段,同时用β-肌动蛋白(β-actin)作为内参照,脑脊液(CSF)阳性标本基因测序分析并总结出临床特征.结果 32例病毒性脑炎脑脊液标本BDV p24基因片段检出率为12.5%(4/32),拷贝数＞102/μl.对其中一份CSF阳性标本测序后,与BDV标准病毒株Ⅴ和马源的BDV病毒株H1766序列比较同源性分别为95.35%和98.84%.在4个位点出现基因突变(nt1649 T→C、nt1656 G→A、nt1670 C→T、nt1676 C→T).该目的基因片段与马源的BDV病毒株亲缘关系最近.阳性脑炎患者主要以精神行为异常为临床特征.结论 贵州省遵义市部分病毒性脑炎的发生与BDV感染有关,主要以精神行为异常为临床特征.     

Borna disease virus infection in racing horses with behavioral and movement disorders

Summary.  Borna disease virus (BDV) is a neurotropic agent with capacity to infect and cause neurological disease in a broad range of warmblooded hosts including horses, sheep, cattle, cats, and possibly also humans. The epidemiology of BDV is largely unknown. However, it is likely that subclinically infected animals may represent potential virus reservoirs. In two groups of Swedish racing horses, one clinically healthy and one consisting of horses with diffuse neurological signs, the BDV seroprevalence was 24.5% and 57.7%, respectively. BDV RNA was detected in peripheral blood mononuclear cells in 8 out of 28 (28.6%) investigated horses, the majority of the BDV RNA-positive horses belonging to the group with neurological signs. There was a close relationship between the Swedish equine BDV isolates and previously reported equine BDVs in Europe. Our results point to an association of BDV infection with atypical disease patterns in horses such as diffuse mental and gait disturbances. These findings may be of importance for the understanding of the epidemiology of BDV infections in animals and man. Accepted September 4, 1998 Received May 27, 1998     

用蛋白印迹试验检测精神分裂症患者血清中博尔纳病病毒-p24抗体的研究

目的：探讨博尔纳病病毒（BDV）感染在我国的流行情况及其与精神分裂症的相关性。方法：在对优化表达的GST－BDV－p24融合蛋白进行特异性鉴定的基础上，通过确定融合蛋白与第一抗体、第二抗体间的最佳反应条件，建立可行的检测BDV－p24特异抗体的蛋白印迹（Western-blot）方法，进而对黑龙江地区精神分裂症患者和正常人对照血清中BDV－p24特异性抗体进行了检测，并通过血清－GST蛋白吸收后W estern-blot实验对阳性血清进行了确认。结果：116例精神分裂症患者中检出BDV－p24阳性血清10例，阳性检出率为8.6%，而正常人血清标本中未检出阳性。结论：我国存在博尔纳病病毒的感染，博尔纳病病毒的感染可能与精神分裂症的发生有关。     

新疆伊犁地区动物脑组织博尔纳病病毒p24基因片段的检测

目的 了解新疆伊犁地区动物脑博尔纳病病毒(Borna disease virus,BDV)感染现状,分析该病毒的种系来源,预防我国BDV大规模暴发流行.方法 采用巢式逆转录实时荧光定量聚合酶链反应(FQ-RT-PCR)的方法,对新疆地区200匹马、75头驴和100只牧羊犬脑组织标本进行BDVp24基因片段的检测.并对阳性标本进行BDV p40基因片段扩增电泳,同时排除GFP-p24和pMD-19质粒污染,最后克隆测序,进行基因同源性和氨基酸序列分析并分析BDV种系发生.结果 3例伊犁马、4例伊犁驴和9例牧羊犬的脑组织标本BDV p24检测阳性,阳性率分别为1.5%、5.3%、9.0%;BDV p40基因片段检测和质粒标准品GFP-p24、pMD-19检测均为阴性;BDV p24扩增产物序列与He/80株的同源性为100%.结论 新疆伊犁地区马、驴和狗可能存在BDV脑内的自然感染,该地区BDV流行株与He/80株存在同源性.     

中国慢性疲劳综合征患者血浆中BDV-p24抗体的检测

目的检测博尔纳病病毒(BDV)对中国慢性疲劳综合征(CFS)患者和健康对照者感染的情况,探讨CFS与BDV感染之间的相关性。方法按美国CDC 1994年标准,搜集来自全国十一省市的CFS患者61例和健康对照73例,使用蛋白印迹法(WB)对其血浆进行BDV-p24抗体检测。结果 病例组有7例为阳性结果(11.48％),对照组均为阴性(0％),统计学处理两组间差异有显著意义(P<0.010)。结论中国CFS患者存在BDV感染,病例组感染率明显高于对照组,CFS与BDV感染存在相关性。     

Adaptation of a sandwich enzyme-linked immunosorbent assay to determine the concentration of bovine leukemia virus p24 and optimal conditions for p24 expression in short-term cultures of peripheral blood mononuclear cells

Bovine leukemia virus (BLV) is a common retroviral infection of cattle. Infection is accompanied by integration of BLV into the host cell genome and is persistent for the life of the individual as is the presence of anti-BLV antibodies. Lymphosarcoma occurs in a small fraction of infected adult individuals but otherwise there is little or no associated disease. Viremia is undetectable, however, BLV is expressed readily once infected cells are cultured in vitro. A sandwich enzyme-linked immunosorbent assay (sELISA) was optimized, using murine monoclonal antibodies, to quantify the major internal structural protein (p24) produced in short-term cultures of peripheral blood mononuclear cells (PBMCs). Optimal production of BLV p24 was achieved utilizing RPMI supplemented with 10% fetal bovine serum (FBS), pH 7, and 5 x 10(6) cells per ml. Cultures were terminated at 24 h. The sELISA was linear between 30 and 900 ng/ml and the limit of detection was 1.2 ng/ml. At three concentrations of p24, intra- and inter-assay coefficients of variation (CV) varied between 9.2 and 13.3 and 5.1 and 12.9%, respectively.     

Borna disease virus infection,a human mental-health risk

This article focuses on human Borna disease virus (BDV) infections, most notably on the development of valid diagnostic systems, which have arisen as a major research issue in the past decade. The significance of a novel modular triple enzyme-linked immunosorbent assay that is capable of specifically measuring anti-BDV antibodies as well as major structural proteins N (p40) and P (p24) in the blood, either as free antigens in the plasma or as antibody-bound circulating immune complexes (CICs), is explained. The impact of CICs and plasma antigen, which indicate periods of antigenemia in the course of BDV infection, along with other infection markers that are still in use is discussed. The review further provides new insight into possible links of BDV to human diseases, summarizing cross-sectional and longitudinal data which correlate acute depression with the presence and amount of antigen and CICs. Moreover, BDV prevalence in healthy people is reevaluated, suggesting that this was previously underestimated. Antiviral efficacy of amantadine, in vivo and in vitro, is outlined as well, with emphasis on wild-type (human and equine) versus laboratory strains. Finally, the pros and cons of the association of BDV with human disease, as detailed in the literature, are critically discussed and related to our data and concepts. This article supports existing correlative evidence for a pathogenic role of BDV infection in particular human mental disorders, in analogy to what has been proven for a variety of animal species.     

No evidence of endemic Borna disease virus infection in Australian horses in contrast with endemic infection in other continents

Summary. Borna disease virus (BDV) is a unique RNA virus that is a cause of neurological disease in horses, sheep and cats. The finding that BDV also infects humans has raised concern related to the impact of infection with this virus. The extent to which BDV may be endemic in geographical regions outside Europe is of interest in management of international movement of animals including horses. Sera from Australian horses (N = 553) sampled in Sydney, New South Wales (NSW), were analysed for BDV antigen, circulating immune complexes (CICs), and antibodies by monoclonal antibody-based ELISAs. One-tenth of the samples were investigated by further antibody tests, namely immunofluorescence (IFA) and a peptide ELISA, as well as for BDV RNA. The study revealed a very low frequency of serological markers that may be associated with exposure to BDV in Australian horses from NSW with a few sera (0.7%) displaying low range positive results in the CIC assay, and no detectable BDV RNA. This pattern is inconsistent with endemic BDV infection and strongly contrasts with the pattern of endemic infection, particularly in Europe.     

家族聚集性慢性乙型病毒性肝炎患者PBMC免疫相关的基因表达谱

目的:应用寡核苷酸基因芯片技术,研究家族聚集性HBV感染者外周血单个核细胞(PBMC)免疫相关基因的表达谱。方法:在一聚集性HBV感染家族中,选取患者5例和患者的正常配偶4例,提取PBMC中的RNA,与涵盖2.2万个EST的寡核苷酸表达谱基因芯片U133A2.0杂交,通过Affymetrix扫描仪和DNT分析软件比较患病组与对照组PBMC免疫相关基因的的表达谱,获得基因的相对表达比值。结果:在2.2万个EST中,初筛出24个免疫相关差异表达基因,7个基因表达上调,17个表达下调。上调的主要是与适应性免疫相关的基因,下调的主要是与固有免疫相关的基因。结论:宿主感染HBV后,可改变其免疫基因表达,固有免疫功能的障碍可能是HBV感染慢性化的主要原因。