Quantitation of herpes simplex virus DNA in cerebrospinal fluid of patients with herpes simplex encephalitis by the polymerase chain reaction

Background: Previous studies have shown the diagnostic utility of qualitative detection of herpes simplex virus (HSV) DNA by the polymerase chain reaction (PCR) in cerebrospinal fluid samples (CSF) from patients with herpes simplex encephalitis (HSE).Objectives: To determine whether quantitation of HSV DNA in CSF could be useful for monitoring efficacy of antiviral therapy and provide prognostic indications.Study design: A quantitative PCR assay using an internal control for evaluation of PCR efficiency and detection of PCR inhibitors was developed and used for retrospective testing of 98 CSF samples from 26 patients with serologically diagnosed HSE during the period 1980–1995.Results: HSV DNA was detected in 36 CSF samples from 23 patients. PCR positivity was 100% for CSF samples collected within 10 days after onset, and 30.4 and 18.7% for samples collected 11–20 and 21–40 days later, respectively. The 3 PCR-negative patients had their first CSF collected 14, 16, and 28 days after onset, respectively. Three of 98 (3.1%) CSF samples were completely or partially inhibitory to PCR. Initial DNA levels were not significantly different in patients with HSE due to either primary or recurrent HSV infection. In addition, they were not related to severity of clinical symptoms nor were predictive of the outcome. A progressive decrease in viral DNA levels was observed both in patients who received acyclovir therapy and in a small number of untreated patients.Conclusions: This study: (i) confirms the high sensitivity of PCR for the diagnosis of HSE; (ii) emphasizes the need for an internal control of amplification to achieve maximal sensitivity and perform reliable quantitation of viral DNA; and (iii) suggests that CSF might not be the best specimen to investigate in studies of the natural history of HSE.     

Serum and cerebrospinal fluid herpes simplex virus type 1 immunoglobulin G and M titers in four cases of herpes simplex encephalitis

Herpes simplex virus type 1 (HSV-1) IgG and IgM ELISA titers were serially determined in serum and cerebrospinal fluid (CSF) samples from 4 patients with HSV-1 encephalitis during a follow-up period of 1-26 months. In 3 out of 4 patients HSV-1 IgM titers raised in CSF during the acute phase of disease, thus allowing differentiation between primary and reactivated forms of HSV-1 encephalitis. HSV-1 IgG titers showed a sharp elevation earlier in serum than in CSF. Specific IgG index documented a large intrathecal production of HSV-1 IgG and their persistence 2 years following clinical onset. The initial trend of serum and CSF specific IgG titer represents a reliable tool for a retrospective diagnosis of HSV-1 encephalitis.     

Proteomics in immunity and herpes simplex encephalitis

The genetic theory of infectious diseases has proposed that susceptibility to life-threatening infectious diseases in childhood, occurring in the course of primary infection, results mostly from individually rare but collectively diverse single-gene variants. Recent evidence of an ever-expanding spectrum of genes involved in susceptibility to infectious disease indicates that the paradigm has important implications for diagnosis and treatment. One such pathology is childhood herpes simplex encephalitis, which shows a pattern of rare but diverse disease-disposing genetic variants. The present report shows how proteomics can help to understand susceptibility to childhood herpes simplex encephalitis and other viral infections, suggests that proteomics may have a particularly important role to play, emphasizes that variation over the population is a critical issue for proteomics and notes some new challenges for proteomics and related bioinformatics tools in the context of rare but diverse genetic defects.     

Coexistence of Alzheimer disease neuropathology with herpes simplex encephalitis.

Several unusual features were observed during routine histopathological confirmation of a clinical diagnosis of Alzheimer's disease (AD) in an 85-year-old, right-handed, married male. The patient presented with a 12-year history of slowly progressive cognitive impairment, which increased in severity just prior to death. Detailed postmortem examination of the frontal lobes revealed a significant number of neuritic plaques and neurofibrillary tangles. Multifocal spongiform encephalopathic changes, mononuclear perivascular infiltrates, subcortical demyelination and gliosis were also found. Of particular interest were well-defined neuronal and astrocytic intranuclear inclusion bodies (Cowdry type I and I), suggestive of viral disease. Electron microscopy, immunohistochemical and immunohistofluorescent studies confirmed a Herpes simplex type I encephalitis (HSV-I). These histological results and the clinical history of progression suggest that reactivation of a latent viral infection may have contributed to the rapid progression of dementia prior to death. The present analysis underscores the fact that multiple etiologic factors may act simultaneously to produce dementia. While one such process may be identified or diagnosed (in the present case AD), it is necessary to be open to the possibility that another mechanism may come into play during the time course of that illness. A differential diagnosis may be difficult when the symptoms of the two disease processes are very similar. Such may be the case if there is reactivation of a previously undiagnosed herpes virus infection. With the development of PCR and in situ hybridization diagnosis will be simplified and more definitive.     

手足口病脑炎患儿脑脊液中细胞因子水平的比较

目的探讨不同病原体(CV-A6、CV-A16、EV-A71)所致手足口病脑炎患儿脑脊液中IL-6、IL-10、TNF-α、IFN-γ水平及意义。方法采用随机抽样的方法选取2018年3—5月在医院感染科收治的HFMD患儿,其中EV-A71组90例,CV-A6组77例,CV-A16组65例,选择同期高热惊厥患儿20例作为对照(高热惊厥组)。患儿入院后1～2 d行腰椎穿刺术,收集脑脊液2 mL,用流式细胞检测术分别检测细胞因子IL-6、IL-10、TNF-α、IFN-γ水平。结果 EV-A71组、CV-A6组、CV-A16组IL-6水平均明显高于高热惊厥组(t分别为6.224、7.579、6.667,P!0.05),且组间差异有统计学意义(F=18.631,P<0.05)。EV-A71组、CV-A6组、CV-A16组IL-10水平均高于高热惊厥组,差异具有统计学意义(t分别为5.387、3.227、3.084,P!0.05),且组间差异有统计学意义(F=17.480,P<0.05)。EV-A71组、CV-A6组、CV-A16组TNF-α水平与高热惊厥组差异均无统计学意义(t=-1.071、1.498、0.400,P>0.05),组间差异有统计学意义(F=6.069,P<0.05)。EV-A71组、CV-A6组、CV-A16组IFN-γ水平均高于高热惊厥组,差异具有统计学意义(t分别为4.718、7.303、8.919,P!0.05),且组间差异有统计学意义(F=16.566,P<0.05)。结论 EV-A71、CV-A6、CV-A16所致重症HFMD患儿脑脊液中IL-6、IL-10、IFN-γ均升高,表明在这3种不同病原体所致HFMD患儿脑炎中,IL-6、IL-10、IFN-γ均起到重要作用。其中,IL-6、IFN-γ明显升高,可作为疾病严重程度的评价指标。     

Biopsy histopathology in herpes simplex encephalitis and in encephalitis of undefined etiology.

The histopathology of herpes simplex encephalitis (HSE) has been described principally from postmortem studies which reveal end-stage disease. Biopsy material, which selects an earlier stage in disease development, has been used principally to isolate virus, identify viral particles, and locate viral antigens. Further, little attention has been paid to the histopathology of biopsies of encephalitis of undefined etiology. In the present study, sections from biopsies which yielded virus and those which were negative for virus were evaluated in a systematic and controlled manner. Biopsies yielding virus were characterized by meningeal inflammation, perivascular infiltrates, and glial nodules. Biopsies which did not yield virus and which failed to reveal another diagnosis were characterized by nonspecific gliosis. Thus the early histiopathology of HSE is characterized by early signs of inflammation in the absence of necrosis and generally differs from biopsies in which virus is not isolated.     

Cerebrospinal fluid miRNA profile in HIV‐encephalitis

MicroRNAs are short non‐coding RNAs that modulate gene expression by translational repression. Because of their high stability in intracellular as well as extracellular environments, miRNAs have recently emerged as important biomarkers in several human diseases. However, they have not been tested in the cerebrospinal fluid (CSF) of HIV‐1 positive individuals. Here, we present results of a study aimed at determining the feasibility of detecting miRNAs in the CSF of HIV‐infected individuals with and without encephalitis (HIVE). We also evaluated similarities and differences between CSF and brain tissue miRNAs in the same clinical setting. We utilized a high throughput approach of miRNA detection arrays and identified differentially expressed miRNAs in the frontal cortex of three cases each of HIV+, HIVE, and HIV? controls, and CSF of 10 HIV‐positive and 10 HIV‐negative individuals. For the CSF samples, the group of HIV+ individuals contained nine cases of HIV‐associated neurological disorders (HAND) and, among those, four had HIVE. All the HIV‐negative samples had non‐viral acute disseminate encephalomyelitis. A total of 66 miRNAs were found differentially regulated in HIV+ compared to HIV? groups. The greatest difference in miRNA expression was observed when four cases of HIVE were compared to five non‐HIVE cases, previously normalized with the HIV‐negative group. After statistical analyses, 11 miRNAs were fund significantly up‐regulated in HIVE. Although more clinical samples should be examined, this work represents the first report of CSF miRNAs in HIV‐infection and offers the basis for future investigation. J. Cell. Physiol. © 2012 Wiley Periodicals, Inc.     

Macrophages and cytokines in the early defence against herpes simplex virus

Herpes simplex virus (HSV) type 1 and 2 are old viruses, with a history of evolution shared with humans. Thus, it is generally well-adapted viruses, infecting many of us without doing much harm, and with the capacity to hide in our neurons for life. In rare situations, however, the primary infection becomes generalized or involves the brain. Normally, the primary HSV infection is asymptomatic, and a crucial element in the early restriction of virus replication and thus avoidance of symptoms from the infection is the concerted action of different arms of the innate immune response. An early and light struggle inhibiting some HSV replication will spare the host from the real war against huge amounts of virus later in infection. As far as such a war will jeopardize the life of the host, it will be in both interests, including the virus, to settle the conflict amicably. Some important weapons of the unspecific defence and the early strikes and beginning battle during the first days of a HSV infection are discussed in this review. Generally, macrophages are orchestrating a multitude of anti-herpetic actions during the first hours of the attack. In a first wave of responses, cytokines, primarily type I interferons (IFN) and tumour necrosis factor are produced and exert a direct antiviral effect and activate the macrophages themselves. In the next wave, interleukin (IL)-12 together with the above and other cytokines induce production of IFN-γ in mainly NK cells. Many positive feed-back mechanisms and synergistic interactions intensify these systems and give rise to heavy antiviral weapons such as reactive oxygen species and nitric oxide. This results in the generation of an alliance against the viral enemy. However, these heavy weapons have to be controlled to avoid too much harm to the host. By IL-4 and others, these reactions are hampered, but they are still allowed in foci of HSV replication, thus focusing the activity to only relevant sites. So, no hero does it alone. Rather, an alliance of cytokines, macrophages and other cells seems to play a central role. Implications of this for future treatment modalities are shortly considered.     

Overproduction of gamma interferon in B/Jas inbred rabbits with herpes simplex virus encephalitis.

Inbred rabbits of the B/Jas strain are highly susceptible to herpes simplex virus type-1 (HSV-1) encephalitis, developing seizures of encephalitis after intravenous injection of the KOS strain of the virus. Anti-viral interferon activity became detectable in the serum just prior to or at the onset of seizures, its level being lower in the serum than in the cerebrospinal fluid. The activity was of gamma interferon, as suggested by the acid instability and the inability of Mx protein induction. An immunohistochemical analysis of the brain tissues of encephalitic rabbits showed that MHC class I antigen was expressed on the microglia cells of inflamed lesions but not on these cells in uninflamed areas. These findings were discussed in correlation with the pathogenesis of herpetic encephalitis in the inbred rabbits.     

Cerebrospinal fluid gamma aminobutyric acid levels in migraine.

Gamma-Aminobutyric acid (GABA) levels in cerebrospinal fluid were measured in seven patients with tension headache and 12 patients with migraine. GABA was detected only during the migraine attack. The results suggest disordered GABA metabolism in migraine.     

Cerebrospinal fluid S-100 protein levels in neurological pathologies

The aim of this paper was to evaluate S-100 concentration in cerebrospinal fluid (CSF) from patients with different neurological disorders, and in subjects with no proven neurological pathology, in order to study possible differences in their protein concentrations. The total number of patient-samples examined was 119 (58 males and 61 females; mean age 35 yrs, 1-79 yrs). Based on the final diagnoses, nine patient groups were studied: a control group, meningitis, acute lymphatic leukemia (ALL), dementia, hydrocephalia, polyneuropathy-motor neuron disease, acute cerebral infarction (ACI), and patients diagnosed with multiple sclerosis. S-100 protein concentrations were measured by the Sangtec 100 two-site immunoradiometric assay. The highest S-100 levels in CSF were found in the dementia group, ACI group, bacterial-fungal and lymphocytic meningitis groups (Kruskal-Wallis test). The S-100 concentrations in these groups were significantly higher compared with the control group (Mann-Whitney U test, p<0.05, p<0.01) and the multiple sclerosis group (p<0.05, p<0.01). No other significant differences were found between groups. Our results suggest that the high protein levels in CSF found in these pathologies may reflect the presence of brain damage. However, the levels need to be considered individually, as they depend on several factors, such as age, severity of brain damage or interval between the onset of brain damage and the taking of the sample.     

Detection of DNA of six human herpesviruses in the cerebrospinal fluid of immunocompetent non‐herpetic acute limbic encephalitis patients

In order to determine whether six other human herpesviruses, aside from herpes simplex virus, are associated with non‐herpetic acute limbic encephalitis in immunocompetent individuals, real‐time PCR was used to detect the DNA of herpesviruses in CSF collected from 61 patients with this form of encephalitis. Five of the human herpesviruses tested were not detected in any of the 61 CSF samples. EBV DNA was detected in one CSF sample. The EBV DNA‐positive patient was a 36‐year‐old woman who presented with fever, headache, mild somnolence, and the typical neuroimaging findings.     

Cerebrospinal fluid cyclic AMP levels in rhesus monkeys: daily fluctuations.

Daily fluctuations in cerebrospinal fluid cyclic AMP levels were found in neurologically intact rhesus monkeys. When animals were maintained in continuous dim light as opposed to a 12L/12D regimen, the fluctuations in cerebrospinal fluid cyclic AMP levels appeared to be reduced. Knowledge of these daily fluctuations may be helpful for protocols of future experiments concerning measurements of CSF cAMP.