运用PCR及ABC—ELISA法早期诊断结论性脑膜炎的评价

收集了４１例结核性脑膜炎（结脑）患者和３０例非结脑患者脑脊液，运用ＰＣＲ法扩增结核蛋白抗原Ｂ上ＤＮＡ序列４１９ｂｐ片段，用ＡＢＣ－ＥＬＩＳＡ检测抗结核蛋白抗体。结脑组ＰＣＲ阳性率７５．７％（３１／４１），抗结核抗体阳性率６３．５％（２６／４１）；对照组ＰＣＲ全部阳性，抗结核抗阳性率１３．３％。ＰＣＲ阳性可在发病第２天查到且在治疗６个月时仍可见ＰＣＲ阳性。随病程延长抗全阳性率上升。运用ＰＣＲ及ＡＢＣ     

运用PCR及ABC－ELISA法早期诊断结核性脑膜炎的评价

收集了４１例结核性脑膜炎（结脑）患者和３０例非结脑患者脑脊液，运用ＰＣＲ法扩增结核蛋白抗原Ｂ上ＤＮＡ序列４１９ｂｐ片段，用ＡＢＣ－ＥＬＩＳＡ检测抗结核蛋白抗体。结脑组ＰＣＲ阳性率７５．７％（３１／４１），抗结核抗体阳性率６３．５％（２６／４１）；对照组ＰＣＲ全部阳性，抗结核抗体阳性率１３．３％。ＰＣＲ阳性可在发病第２天查到且在治疗６个月时仍可见ＰＣＲ阳性。随病程延长抗体阳性率上升。运用ＰＣＲ及ＡＢＣ－ＥＬＩＳＡ法早期诊断结脑快速而敏感，两法同时应用可提高阳性率。     

3D—CTA,MRA和DSA对脑动静脉畸形成像的对照性研究

目的：评价数字减影血管造影（ＤＳＡ）、三维计算机断层扫描血管造影（３Ｄ－ＣＴＡ）和磁共振血管成像技术（ＭＲＡ）对于脑动静脉畸形（ＡＶＭ）甩像的价值。方法 ４３例ＡＶＭ前瞻性对照研究,结果 ３Ｄ－ＣＴＡ阳性率１００％,与ＤＳＡ比较;供血动脉的来源,数目和形态１００％吻合,引流静脉３３例完全吻合（７８．６％）,９例部分吻合（２１．４％）;病灶部位、大小以及深度１００％吻合。ＭＲＡ阳性率９５．８％,１例假阴性（４．２％）。与ＤＳＡ影像比较,供血动脉１７例完全吻合（７０．８％）,６例部分吻合（２５．０％）;引流静脉１４例完全吻合（５８．３％）,９例部分吻合（３７．５％）,病灶部位、大小以及深度９８．５％吻合。结论 ＤＳＡ是脑ＡＶＭ成像的“金标准”。３Ｄ－ＣＴＡ和ＭＲＡ技术的发展为ＡＶＭ影像不诊断提供了高准确性,无创性的     

PCR DNA扩增、抗原、特异性抗体联合检测对结核性脑膜炎的诊断研究

应用多聚酶链反应技术（ＰＣＲ）体外扩增脑脊液中结核分枝杆菌ＤＮＡ，酶斑免疫结合技术检测结核杆菌蛋白抗原及ＡＢＣ－ＥＬＩＳＡ法检测相应抗体联合检测诊断结核性脑膜炎，阳性率分别为：８３．７％（３６／４３），７６．７％（３３／４３），７４．４％（３２／４３）。讨论三者间发病不同期的关系，三者有阳性交叉现象。     

重症肌无力IgG型及IgM型抗突触前膜抗体检测

利用β－银环蛇毒素（β－ｂｕｎｇａｒｔｏｘｉｎβ－ＢＧＴ）特异性与突触前膜不可逆结合的特性提纯突触前膜相应蛋白受体为抗原，用ＥＬＩＳＡ检测４０例重症肌无力病人，发现重症肌无力病人血中存有抗突触前膜抗体。ＩｇＧ型阳性率（６０％）较ＩｇＭ型阳性率（２７．５％）高。提示重症肌无力病人突触前膜也受自身抗体的损伤。同时本实验还检测（抗乙酰胆碱受体ＡＣｈＲ抗体）作为对照     

单纯疱疹病毒性脑炎脑脊液三项检查的临床评价

为评价目前常用的单纯疱疹病毒性脑炎（ＨＳＥ）脑脊液（ＣＳＦ）３项检查方法的临床应用价值，作者按照平行试验的原则，对４３例ＨＳＥ患者同时进行了ＣＳＦ单纯疱疹病毒抗原（ＨＳＶ－Ａｇ）、抗体（ＨＳＶ－Ａｂ）和ＤＮＡ（ＨＳＶ－ＤＮＡ）检测。结果显示：ＨＳＶ－Ａｂ与ＨＳＶ－ＤＮＡ检测的阳性率高于ＨＳＶ－Ａｇ，分别为８６．０％、７６．７％和４８．８％；４３例患者中２项以上阳性者为３６例，占８３．７％。作者认为：对临床疑诊的所谓“散发性脑炎”患者均应同时进行ＣＳＦＨＳＶ３项检测，如２项或３项阳性者可确诊为ＨＳＥ，１项阳性者应高度怀疑为ＨＳＥ，并应早期使用无环鸟苷等有效药物治疗。     

格林—巴利综合征患者血清中的抗糖脂抗体

采用固相酶联免疫吸附法对３５例急性期格林－巴利综合征（ＧＢＳ）患者、２８例其他神经系统疾病患者和３０例健康体检者的血清中抗硫脂抗体，抗ＧＱ１ｂ及抗ＧＭ１抗体进行检测。结果：ＧＢＳ患者血清中抗硫脂ＩｇＭ抗体、抗ＧＱ１ｂＩｇＧ抗体和抗ＧＭ１ＩｇＧ抗体阳性率分别为３４％、１１％和３１％，均显著高于正常对照组。５６％的抗硫脂抗体阳性患者均有不同程度感觉障碍，而抗硫脂抗体阴性患者仅１６％（Ｐ＜０．０５）。５例有眼肌运动障碍的ＧＢＳ患者中，４例抗ＧＱ１ｂＩｇＧ抗体阳性，无眼肌麻痹的ＧＢＳ患者无１例抗ＧＱ１ｂ抗体阳性。提示不同的抗糖脂抗体可能在ＧＢＳ发病过程中起不同的作用。     

淫斑点免疫渗滤试验诊断单纯疱疹病毒性脑炎

采用斑点免疫渗滤试验（ＤＩＦＡ）对苏州地区１７０例散发性脑炎患者２４０份脑脊液（ＣＳＦ）标本中单纯疱疹病毒抗原（ＨＳＶ－Ａｇ）进行检测，结果表明１７０例散发性脑炎中，经该项检测而能明确单纯疱疹病毒性脑炎（ＨＳＥ）诊断者４２例（２４．７％）；２４０份ＣＳＦ标本中，ＨＳＶ－Ａｇ阳性率为３５．４％，而其中４２例ＨＳＥ患者的９６份ＣＳＦ阳性率为８８．５％，两者差异有非常显著意义（Ｐ〈０．００１）。将ＤＩＦ     

重症肌无力抗突触受体抗体的检测及其意义

目的：探讨一种阳性率更高的实验指标，以提高重症肌无力的实验室诊断率。方法：将α一银环蛇毒素和β－银环蛇毒素混合一起包被酶标板，结合随后加的肌肉提取液中相应蛋白质为抗原，以ＡＢＣ－ＥＬＩＳＡ法检测重症肌无力病人血清中混合的抗乙酰胆碱受体抗体（ＡｃｈＲａｂ）和抗突触前膜受体抗体（ＰｓｍＲａｂ），称为抗突触受体抗体。结果：７５名正常人均阴性。８０例临床对照组中，除２例运动神经元疾病和１例多发性肌炎阳性外，其余均阴性。本组１２２例重症肌无力病人中９８例阳性（８０．３％，其中全身型阳性８７．５％），阳性率明显高于单独检测ＡｃｈＲａｂ，（Ｐ＜０．００１）。结论：检测抗突触受体抗体可作为重症肌无力诊断的一个阳性率较高的较好的实验室指标。     

格林－巴利综合征患者血清髓鞘脂抗体检测及其临床意义

应用ＥＬＩＳＡ方法对２８例格林－巴利综合征（ＧＢＳ）患者和３０例正常对照者、３０例疾病对照者进行血清抗髓鞘脂抗体检测，并对其中５例进行动态观察。结果发现：ＧＢＳ患者血清抗髓鞘脂抗体水平增高，与正常和疾病对照组相比有非常显著性差异（Ｐ＜０．０１）。ＧＢＳ患者髓鞘脂抗体出现的阳性率为２１．４％，ＧＢＳ患者髓鞘脂抗体水平下降与临床症状的改善相一致。     

The antigenic binding sites of autoantibodies to factor XII in patients with recurrent pregnancy losses

Recently, numerous studies have suggested an association between factor XII (FXII) deficiency and recurrent pregnancy losses, and between autoantibodies to FXII and recurrent pregnancy losses. Autoantibodies to FXII rather than FXII deficiency may be a risk factor for recurrent pregnancy losses. To know the pathogenesis of autoantibodies to FXII, epitope mapping study was done. Seventeen anti-FXII antibody positive recurrent pregnancy loss patients were chosen for this study. We used synthetic peptides in inhibition and direct binding studies to identify the antigenic binding site of autoantibodies to FXII. Among plasmas from 17 recurrent pregnancy loss patients who were positive for autoantibodies to FXII, 13 patients (76.5%) recognized amino acids 1-30, the amino-terminal heavy chain region that is known as factor XII binding site to platelet glycoprotein Ibalpha.     

The prevalence of autoantibodies among right and left handed schizophrenic patients and control subjects.

Sera from schizophrenic patients (n = 186) and healthy control subjects (n = 346) were tested for the presence of seven common autoantibodies by standard immunological methods. The association between handedness and autoantibodies was tested in a multi-way contingency table using a log-linear model. For men, but not women, nondextrals (patients and controls) were twice as likely to test positive for autoantibodies than dextrals (p = 0.0002). Although more women (33%) than men (24%) tested positive for autoantibodies, handedness was not a distinguishing factor among women. These data suggest that sinistrality and gender are associated with autoantibodies in a subgroup of schizophrenic patients and healthy control subjects.     

Detection of plasma autoantibodies to brain tissue in young children with and without autism spectrum disorders

Autism spectrum disorders (ASDs) are characterized by impaired language and social skills, often with restricted interests and stereotyped behaviors. A previous investigation of blood plasma from children with ASDs (mean age = 5½ years) demonstrated that 21% of samples contained autoantibodies that reacted intensely with GABAergic Golgi neurons of the cerebellum while no samples from non-sibling, typically developing children showed similar staining (Wills et al., 2009). In order to characterize the clinical features of children positive for these autoantibodies, we analyzed plasma samples from children enrolled in the Autism Phenome Project, a multidisciplinary project aimed at identifying subtypes of ASD. Plasma from male and female children (mean age = 3.2 years) was analyzed immunohistochemically for the presence of autoantibodies using histological sections of macaque monkey brain. Immunoreactivity to cerebellar Golgi neurons and other presumed interneurons was observed for some samples but there was no difference in the rate of occurrence of these autoantibodies between children with ASD and their typically developing peers. Staining of neurons, punctate profiles in the molecular layer of the dentate gyrus, and neuronal nuclei were also observed. Taken together, 42% of controls and subjects with ASD demonstrated immunoreactivity to some neural element. Interestingly, children whose plasma reacted to brain tissue had scores on the Child Behavior Checklist (CBCL) that indicated increased behavioral and emotional problems. Children whose plasma was immunoreactive with neuronal cell bodies scored higher on multiple CBCL scales. These studies indicate that additional research into the genesis and prevalence of brain-directed autoantibodies is warranted.     

No increase of serum autoantibodies during therapy with recombinant human interferon-β1a in relapsing-remitting multiple sclerosis

Objectives - The present investigation was aimed at establishing whether interferon (IFN)-β would induce the synthesis of autoantibodies in patients affected by multiple sclerosis (MS). Materials and methods - The titres of different autoantibodies were measured in a group of 68 relapsing-remitting MS patients before and during treatment with human recombinant IFN-β_la (3 MIU or 9 MIU subcutaneously 3a week). ANA, anti-thyroid, anticardiolipin serum autoantibodies were assayed in all cases: when patients were found positive to ANA>1: 40, they were also tested for anti-DNA and anti-ENA antibodies. Results - No increase was found in autoantibodies synthesis during 6 months of r-hIFNβ_la therapy, either at low or high dosages. The percentage of patients positive to different types of autoantibodies varied between 0 and 29%, which are values similar to those already reported in untreated MS patients. Conclusion - Our data indicate that the short-term use of IFN-β_1a in MS is safe in terms of the induction of humoral autoimmune responses: however, further follow-up is needed to confirm these findings during long-term treatments.     

Cerebrospinal fluid anti-cerebellar soluble lectin antibodies in human immunodeficiency virus type 1 infection.

Cerebrospinal fluid samples from 14 human immunodeficiency type 1 (HIV-1) seropositive patients in various stages of HIV infection were tested for the presence of autoantibodies to an endogenous manose-binding protein, the cerebellar soluble lectin (CSL), which has recently been found to be detected in a high proportion of patients with multiple sclerosis. An immunoblotting test was used with rat CSL as antigen. Seven patients were positive for anti-CSL and seven were negative. The seven anti-CSL-positive patients had signs of intrathecal immunoglobulin G production measured as an elevated IgG index, while the seven anti-CSL-negative patients had a normal IgG index. There was no apparent relation between infectious stage and the presence of anti-CSL. Immunological reactions such as anti-CSL autoantibodies may be a similar pathogenic mechanism in HIV and multiple sclerosis brain disease.     

Anti-GD2-like IgM autoreactivity in multiple sclerosis patients

Seric IgM autoreactivity in 100 multiple sclerosis (MS) and 106 control (70 of whom had other neurological diseases) patients was assessed either by immunohistochemistry on normal human CNS tissue or to GD2, GD1a, GD3 by ELISA and thin layer chromatography (TLC) techniques. By double immunohistochemistry, we found that 44% of the total MS population showed seric IgM reactivity to oligodendrocytes and myelin, this finding being particularly frequent in patients with secondary progressive MS. In the non-MS cohort, positive signals were seen only in one patient. In all cases, extraction of lipids from CNS sections abolished the immunoreactivity. Among the gangliosides investigated by ELISA, anti-GD2-like IgM autoantibodies were detected in the serum of 30% of MS patients, a subgroup of whom (below 10%) reacted also with GD1a and/or GD3. More than 85% of MS cases with anti-GD2-like IgM immunoreactivity by ELISA showed also IgM antioligodendrocyte/myelin staining by immunohistochemistry. However, no immunostaining in MS sera was observed when gangliosides were resolved by TLC. A positive correlation with neurological disability was observed, as the Expanded Disability Status Scale of MS patients with anti-GD2-like IgM autoreactivity by ELISA was significantly worse than seronegative MS cases. The results of the present study enforce the role of glycolipids as potential autoantigens and of IgM autoantibodies in MS pathogenesis.     

Glutamic acid decarboxylase autoantibodies in controlled and uncontrolled epilepsy: a pilot study

There have been anecdotal reports of raised glutamic acid decarboxylase (GAD) autoantibodies in patients with refractory epilepsy. We measured serum GAD autoantibodies in 105 patients with idiopathic or symptomatic epilepsy. There was no significant difference in the absolute titre of GAD autoantibody between patients with controlled and uncontrolled epilepsy. However, four female patients with uncontrolled epilepsy had levels that were over three times above the highest detected in the seizure-free group, three of whom also tested positive for pancreatic islet cell antibodies. Larger scale studies, perhaps comparing different epilepsy syndromes, are required to determine the exact clinical role of GAD autoantibodies in epilepsy.     

Autoantibodies against brain septal region antigens specific to unmedicated schizophrenia?

Health et al. (1989) reported that serum from 96% of unmedicated schizophrenic patients contained IgG autoantibodies specific for the septal region of rhesus monkey brain, compared with 0% of nonschizophrenic control subjects and 6% of schizophrenic patients who were on neuroleptic medication. Using the same technique of crossed immunoelectrophoresis, we have tried to replicate this finding. In contrast to the original report, we observed "positive" precipitin arcs with IgG concentrates from all 14 serum samples tested. The failure of immunoelectrophoretic methods to provide convincing evidence of pathogenic autoantibodies in schizophrenia in no way detracts from the hypothesis that autoimmune processes are involved in some forms of schizophrenia. Such methods have not proved useful in established autoimmune diseases such as Graves' disease and myasthenia gravis in which the pathogenic autoantibodies against cell-surface receptors can only be detected by assays which measure functional interactions with such receptors.     

Activating autoantibodies against G protein-coupled receptors in narcolepsy type 1

Study objectivesNarcolepsy type 1 is a rare hypersomnia of central origin, which is caused by loss of hypothalamic neurons that produce the neuropeptides hypocretin-1 and -2. Hypocretin-containing nerve terminals are found in areas known to play a central role in autonomic control and in pain signaling. Cholinergic M2 receptors are found in brain areas involved with the occurrence of hallucinations and cataplexy. In addition to classical symptoms of narcolepsy, the patients suffer frequently from autonomic dysfunction, chronic pain, and hypnagogic/hypnopompic hallucinations. We aimed to test whether narcolepsy type 1 patients have autoantibodies against autonomic β2 adrenergic receptor, M2 muscarinic receptors, or nociception receptors.MethodsWe tested the serum of ten narcolepsy type 1 patients (five female) for activating β2 adrenergic receptor autoantibodies, M2 muscarinic receptor autoantibodies, and nociception receptor autoantibodies.ResultsTen of ten patients were positive for muscarinic M2 receptor autoantibodies (P < 0.001), 9/10 were positive for autoantibodies against nociception receptors (P < 0.001), and 5/10 were positive for β2 adrenergic receptor autoantibodies (P < 0.001).ConclusionsNarcolepsy type 1 patients harbored activating autoantibodies against M2 muscarinic receptors, nociception receptors, and β2 adrenergic receptors. M2 receptor autoantibodies may be related to the occurrence of cataplexy and, moreover, hallucinations in narcolepsy since they are found in the same brain areas that are involved with these symptoms. The occurrence of nociception receptor autoantibodies strengthens the association between narcolepsy type 1 and pain. The connection between narcolepsy type 1, autonomic complaints, and the presumed cardiovascular morbidity might be associated with the occurrence of β2 adrenergic receptor autoantibodies. On the other hand, the presence of the autoantibodies may be secondary to the destruction of the hypocretin pathways.     

CD5 B cells and CD48 T cells in neuroimmunological diseases

Using 2- and 3-colour FACS analysis we found increased levels of fetal-type CD5+ B cells and CD4-8- T cells in cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) and aseptic meningitis (AM) compared to control probands with muscular tension headache (TH). Similar differences were found for CD5+ B cells in peripheral blood, but at lower levels. CD4-8- T cells in blood exceeded those in CSF in all patient groups, with the exception of relapsing remitting MS, revealing the highest values in AM. There was a positive correlation between CD4-8- T cells and T cell receptor (TCR) gamma delta bearing T cells in blood and CSF. The double-negative T cells exceeded the TCR gamma delta T cells by about 1%. A positive correlation between CD5+ B cells and CD4-8- T cell level in CSF was found in MS and AM, but not in TH, nor in blood of any patient group. HLA-DR expression was lower in CD5+ B cells than in CD5- B cells. We conclude that fetal-type lymphocytes are enriched in CSF compartment of patients with inflammatory diseases of the central nervous system, irrespective of autoimmune mechanisms involved, but the function of CD5+ B cells is mainly to produce the autoantibodies.