Antigen In Cerebrospinal Fluid Research Articles

Meningeal cryptococcosis and SARS-CoV-2 infection in people living with HIV/AIDS.

Fungal infections in patients with COVID-19 was one of the most debated topics during the pandemic. To analyze the clinical characteristics and evolution of people living with HIV/AIDS and coinfection with cryptococcus and COVID-19 (group A) or without it (group B). This is an analytical and retrospective study. We reviewed medical records of patients with meningeal cryptococcosis between April 2020 and May 2021. We studied 65 people living with HIV/AIDS and with cryptococcosis infection diagnosed from April 2020 to May 2021. Fifteen patients with HIV/AIDS suffered from cryptococcosis and COVID-19, and out of these, 14 presented meningitis (group A), while 28 suffered from meningeal cryptococcosis, but did not have COVID-19 (group B). No statistically significant differences were observed between the two groups (A and B) considering: intracranial hypertension, presence of Cryptococcus antigens in cerebrospinal fluid, sensorium deterioration or mortality. The detection of Cryptococcus antigens in serum by lateral flow assay was highly effective to rapidly diagnose cryptococcosis in patients with HIV/AIDS who also developed COVID-19. Patients of both groups consulted for cryptoccocosis sometime after, in comparison with the pre-pandemic cases related to this infection.

Arachnoid granulations are lymphatic conduits that communicate with bone marrow and dura-arachnoid stroma.

Arachnoid granulations (AG) are poorly investigated. Historical reports suggest that they regulate brain volume by passively transporting cerebrospinal fluid (CSF) into dural venous sinuses. Here, we studied the microstructure of cerebral AG in humans with the aim of understanding their roles in physiology. We discovered marked variations in AG size, lobation, location, content, and degree of surface encapsulation. High-resolution microscopy shows that AG consist of outer capsule and inner stromal core regions. The fine and porous framework suggests uncharacterized functions of AG in mechanical CSF filtration. Moreover, internal cytokine and immune cell enrichment imply unexplored neuroimmune properties of these structures that localize to the brain-meningeal lymphatic interface. Dramatic age-associated changes in AG structure are additionally identified. This study depicts for the first time microscopic networks of internal channels that communicate with perisinus spaces, suggesting that AG subserve important functions as transarachnoidal flow passageways. These data raise new theories regarding glymphatic-lymphatic coupling and mechanisms of CSF antigen clearance, homeostasis, and diseases.

Investigation of Concurrent Pneumococcal Meningitis in Two Children Attending the Same Day-Care Center

Only a few clusters of invasive pneumococcal disease have been described globally in children, and most of these cases occurred before pneumococcal vaccination implementation. Two unusual cases of pneumococcal meningitis, occurring in the same daycare center over a 3-day period, were reported. Both cerebrospinal fluid (CSF) were sent to the National reference center for pneumococci. In addition, we decided to perform a pneumococcal carriage study on all children and staff of the daycare center to analyze the pneumococcal serotypes circulating in this DCC and to discuss an antibiotic chemoprophylaxis. CSF culture was positive for pneumococcus, and serotype 25A was identified by latex agglutination. The second case had negative CSF culture, but CSF antigen test and gene amplification results were positive for Streptococcus pneumoniae. Serotype 12F was identified by using molecular biology. The absence of correlation between these strains was confirmed by multi-locus sequence typing. In the carriage study, we included 29 children (median age 1.9 years, interquartile range 1.4–2.5) and 10 adults. Among the children, 24 carried Streptococcus pneumoniae (83%). The main serotypes isolated were 23A for 6 children and 25A for 5 children; serotypes were non-typeable for 3 children. Only 1 of 10 adults tested carried Streptococcus pneumoniae (serotype 12F). Despite this temporo-spatial pattern, the cases were unrelated and not due to carriage of a particular serotype. No specific action has been taken for the other children attending this DCC, and no other case of bacterial meningitis occurred.

Viral Antigen and Inflammatory Biomarkers in Cerebrospinal Fluid in Patients With COVID-19 Infection and Neurologic Symptoms Compared With Control Participants Without Infection or Neurologic Symptoms

Neurologic symptoms are common in COVID-19, but the central nervous system (CNS) pathogenesis is unclear, and viral RNA is rarely detected in cerebrospinal fluid (CSF). To measure viral antigen and inflammatory biomarkers in CSF in relation to neurologic symptoms and disease severity. This cross-sectional study was performed from March 1, 2020, to June 30, 2021, in patients 18 years or older who were admitted to Sahlgrenska University Hospital, Gothenburg, Sweden, with COVID-19. All patients had CSF samples taken because of neurologic symptoms or within a study protocol. Healthy volunteer and prepandemic control groups were included. SARS-CoV-2 infection. Outcomes included CSF SARS-CoV-2 nucleocapsid antigen (N-Ag) using an ultrasensitive antigen capture immunoassay platform and CSF biomarkers of immune activation (neopterin, β2-microglobulin, and cytokines) and neuronal injury (neurofilament light protein [NfL]). Forty-four patients (median [IQR] age, 57 [48-69] years; 30 [68%] male; 26 with moderate COVID-19 and 18 with severe COVID-19 based on the World Health Organization Clinical Progression Scale), 10 healthy controls (median [IQR] age, 58 [54-60] years; 5 [50%] male), and 41 patient controls (COVID negative without evidence of CNS infection) (median [IQR] age, 59 [49-70] years; 19 [46%] male) were included in the study. Twenty-one patients were neuroasymptomatic and 23 were neurosymptomatic (21 with encephalopathy). In 31 of 35 patients for whom data were available (89%), CSF N-Ag was detected; viral RNA test results were negative in all. Nucleocapsid antigen was significantly correlated with CSF neopterin (r = 0.38; P = .03) and interferon γ (r = 0.42; P = .01). No differences in CSF N-Ag concentrations were found between patient groups. Patients had markedly increased CSF neopterin, β2-microglobulin, interleukin (IL) 2, IL-6, IL-10, and tumor necrosis factor α compared with controls. Neurosymptomatic patients had significantly higher median (IQR) CSF interferon γ (86 [47-172] vs 21 [17-81] fg/mL; P = .03) and had a significantly higher inflammatory biomarker profile using principal component analysis compared with neuroasymptomatic patients (0.54; 95% CI, 0.03-1.05; P = .04). Age-adjusted median (IQR) CSF NfL concentrations were higher in patients compared with controls (960 [673-1307] vs 618 [489-786] ng/L; P = .002). No differences were seen in any CSF biomarkers in moderate compared with severe disease. In this study of Swedish adults with COVID-19 infection and neurologic symptoms, compared with control participants, viral antigen was detectable in CSF and correlated with CNS immune activation. Patients with COVID-19 had signs of neuroaxonal injury, and neurosymptomatic patients had a more marked inflammatory profile that could not be attributed to differences in COVID-19 severity. These results highlight the clinical relevance of neurologic symptoms and suggest that viral components can contribute to CNS immune responses without direct viral invasion.

Identification of bacterial aetiology in acute meningitis.

The lack of rapid and sensitive test remains a key issue in diagnosing meningitis and affordability impedes using the molecular techniques. However, conventional PCR is currently becoming more affordable. Optimize and establish a multiplex PCR and to compare the above PCR to Cerebrospinal fluid (CSF) culture and antigen detection in sensitivity and specificity for the detection of bacterial meningitis. CSF specimens were collected from patients with suspected acute meningitis admitted to Teaching Hospital, Peradeniya from December 2016 to March 2017. A multiplex PCR was used to detect Neisseria meningitides, Streptococcus pneumonia and Haemophilus influenzae. Eighty specimens of CSF were collected during the study period. The mean duration to sample collection was 4.78(SD 2.6) from the onset of symptoms. None of the samples given positive CSF culture. CSF antigen detection was performed on 50 specimens and all were negative. Of the total samples, eight yielded positive PCR results. In two of the positives, the full report was normal, one was suggestive of viral aetiology and five were suggestive of bacterial aetiology. Three were positive for S.pneumoniae and five for H.influenzae. positive PCR results were associated with a shorter time gap between hospitalization and sample collection and a larger CSF volume. Findings of the study highlight the usefulness and recommendation of multiplex PCR in the diagnosis of pathogens causing acute bacterial meningitis. Collection of an adequate volume of CSF early in the illness, without delay may improve the diagnosis.

An observational study on the association of anti-thyroid autoantibodies with clinical, EEG, MRI, FDG-PET, cerebrospinal fluid and anti-neuronal antibody findings in 530 patients with schizophreniform and affective disorders

IntroductionAlthough the link between autoimmune thyroiditis and mental illnesses is well established, the precise underlying pathophysiology and the influence of anti-thyroid antibodies on diagnostic findings require further research. Patients and MethodsA total of 530 patients with schizophreniform and affective syndromes were screened for anti-thyroid antibodies against thyroid peroxidase (TPO), thyroglobulin (TG), and thyroid-stimulating hormone receptor (TSH-R). The patient group analyzed here is a patient subgroup of a previously published cohort (Endres et al., 2020, Translational Psychiatry). The anti-thyroid antibody positive (N = 91) and negative (N = 439) patients were compared in terms of various clinical parameters, routine cerebrospinal fluid (CSF) findings, and the number of positive anti-neuronal antibodies in serum and/or CSF, as well as electroencephalography (EEG), magnetic resonance imaging (MRI), and [18 F]fluorodeoxyglucose positron emission tomography (FDG-PET) findings. ResultsAnti-TPO antibodies were increased in 17%, anti-TG antibodies in 15%, and anti-TSH-R antibodies in 2% of all patients. In CSF, higher protein concentrations (p = 0.018) and albumin quotients (p = 0.008) were found in the anti-thyroid antibody positive patient group. Also, there were more patients with elevated age-corrected albumin quotients in this group (p = 0.031). FDG-PET hypometabolism was significantly more frequent and the number of positive anti-neuronal intracellular antibodies was significantly higher in patients with anti-thyroid antibodies (p = 0.048, N = 29 and p = 0.032, N = 497 respectively). In addition, there was a trend for higher white blood cell (WBC) counts in all patients with anti-thyroid antibodies (p = 0.090). In the patient subgroup with anti-TPO antibodies this difference was statistically significant (p = 0.027). No relevant differences were found in the other CSF routine parameters, the number of anti-neuronal antibodies against cell surface antigens in serum and/or CSF, EEG and MRI findings. DiscussionThe present study provides evidence of impaired blood CSF barrier (BCSFB) function in patients with anti-TPO and anti-TG antibodies. An influence of anti-TG antibodies on BCSFB structures has been shown in previous laboratory studies, which reported that the antibodies bind to vascular smooth muscle cells. Due to BCSFB breakdown anti-thyroid antibodies might lead to increased autoimmune susceptibility. The alterations in the FDG-PET, WBC count, and anti-neuronal antibody findings against intracellular structures indicate that it could be useful to extend diagnostic investigations in patients with anti-thyroid antibodies. Further studies should investigate whether anti-thyroid antibodies can also act as “drivers of disease”.

Clinical features of bacterial meningitis among hospitalised children in Kenya

BackgroundDiagnosing bacterial meningitis is essential to optimise the type and duration of antimicrobial therapy to limit mortality and sequelae. In sub-Saharan Africa, many public hospitals lack laboratory capacity, relying on clinical features to empirically treat or not treat meningitis. We investigated whether clinical features of bacterial meningitis identified prior to the introduction of conjugate vaccines still discriminate meningitis in children aged ≥60 days.MethodsWe conducted a retrospective cohort study to validate seven clinical features identified in 2002 (KCH-2002): bulging fontanel, neck stiffness, cyanosis, seizures outside the febrile convulsion age range, focal seizures, impaired consciousness, or fever without malaria parasitaemia and Integrated Management of Childhood Illness (IMCI) signs: neck stiffness, lethargy, impaired consciousness or seizures, and assessed at admission in discriminating bacterial meningitis after the introduction of conjugate vaccines. Children aged ≥60 days hospitalised between 2012 and 2016 at Kilifi County Hospital were included in this analysis. Meningitis was defined as positive cerebrospinal fluid (CSF) culture, organism observed on CSF microscopy, positive CSF antigen test, leukocytes ≥50/μL, or CSF to blood glucose ratio <0.1.ResultsAmong 12,837 admissions, 98 (0.8%) had meningitis. The presence of KCH-2002 signs had a sensitivity of 86% (95% CI 77–92) and specificity of 38% (95% CI 37–38). Exclusion of ‘fever without malaria parasitaemia’ reduced sensitivity to 58% (95% CI 48–68) and increased specificity to 80% (95% CI 79–80). IMCI signs had a sensitivity of 80% (95% CI 70–87) and specificity of 62% (95% CI 61–63).ConclusionsA lower prevalence of bacterial meningitis and less typical signs than in 2002 meant the lower performance of KCH-2002 signs. Clinicians and policymakers should be aware of the number of lumbar punctures (LPs) or empirical treatments needed for each case of meningitis. Establishing basic capacity for CSF analysis is essential to exclude bacterial meningitis in children with potential signs.

CTNI-06. SINGLE-CENTER REAL-WORLD ANALYSIS ON CEREBROSPINAL FLUID IN PATIENTS WITH LEPTOMENINGEAL METASTASES OF NON-SMALL CELL LUNG CANCER

Abstract OBJECTIVE To investigate the clinical values of real-world cerebrospinal fluid (CSF) examination in the diagnosis and prognosis of leptomeningeal metastases (LM) in the patients with non-small cell lung cancer (NSCLC). METHODS The clinical manifestations of patients diagnosed with NSCLC-LM in Guangdong Sanjiu Brain Hospital from 2010 until 2019 and the results from the routine, biochemical and cytologic examinations of their CSF were analyzed retrospectively. Moreover, the effects of different indicators on the prognosis of patients were determined by survival analysis. RESULTS Fifty-eight of the 80 patients underwent CSF examination, and the median values of the initial CSF pressure measured by lumbar puncture was 255mmH2O (60-350mmH2O). Tumor cells were found by the cytologic examination of CSF in 56 of the 58 cases (96.6%). The median levels of protein, glucose, chloride, lactate dehydrogenase (LDH) and carcinoembryonic antigen (CEA) in CSF were 0.4g/L (0–8.3g/L), 2.7mmol/L (0–11.9mmol/L), 120.7mmol/L (0–140.5mmol/L), 21.3U/L (0–221.5U/L) and 31.4ng/ml (0–784.2ng/ml), respectively. Sixteen patients underwent gene detection, and the results showed that there were 4 cases of wild type and 12 cases of mutation type. The survival analysis revealed that the median overall survival (mOS) after being diagnosed with LM, patients in LDH≤37U/L group and those in LDH &amp;gt;37U/L group were 8.0 months, 23.7 months and 4.6 months (P=0.039), respectively, which indicated that the LDH level in CSF was the independent prognostic factor for the survival. CONCLUSION Using the cytologic examination of CSF in patients with LM to detect tumor cells is conducive to the early diagnosis of LM. The high CSF pressure in patients with LM is one of the reasons for the neurologic signs and symptoms. The gene mutations found in the CSF in some patients can be used to guide the further systematic treatment. The LDH level in CSF is the independent prognostic factor affecting the survival.

Pathogen-specific risk of seizure in children with moderate-to-severe diarrhoea: Case control study with follow-up.

To determine the pathogen-specific risk of seizure in under-five children hospitalised with moderate-to-severe diarrhoea (MSD) in rural settings. This was a prospective case-control study with follow-up, conducted in a sentinel facility of Global Enteric Multicenter Study in Mirzapur, a rural community of Bangladesh between 2007 and 2010. Children aged 0-59months who presented with MSD and seizure constituted the cases whereas those who did not have seizure comprised the controls. MSD was defined if the episodes were associated with dehydration or dysentery or required hospitalisation with diarrhoea or dysentery. All enrolled children were followed up at home within 50-90days of enrolment. A total of 64 cases and 128 randomly selected controls formed the analysable dataset. The result of logistic regression analysis after adjusting for potential confounders revealed that shigellosis (Shigella species, OR=5.34, 95% CI=2.37-12.04) particularly S. flexneri (OR=3.34, 95% CI=1.48-7.57), S. flexneri 6 (OR=23.24, 95% CI=2.79-193.85), S. sonnei (OR=6.90, 95% CI=2.34-19.85); norovirus (OR=6.77, 95% CI=1.69-27.11), fever (OR=16.75, 95% CI=1.81-154.70) and loss of consciousness (OR=35.25, 95% CI=1.71-726.20) were the independent risk factors for seizure in MSD children. At enrolment, cases had lower WHZ (P=0.006) compared to their peers, follow-up anthropometrics showed significant improvement in WHZ (P<0.001) and WAZ (P<0.05), whereas deterioration in HAZ (P<0.001) in both cases and controls. Childhood MSD episodes particularly due to Shigella and norovirus are often associated with seizure. Prompt identification and appropriate management of children with shigellosis may reduce occurrence and adverse consequences of seizure linked with MSD.

Cryptococcosis in HIV – Infected Hospitalized Patients in Latvia

Aims: To determine the prevalence of cryptococcal infection among HIV hospitalized patients, to evaluate clinical characteristics and outcomes in Latvia.&#x0D; Study Design: Cross-sectional study.&#x0D; Place and Duration of Study: Riga Eastern Clinical University Hospital, Latvian Center of Infectology, between January 2014 and February 2017.&#x0D; Methodology: We conducted the study reporting demographics, epidemiological (age, sex, clinical aspects, paraclinical results (cryptococcal antigen in cerebrospinal fluid, serum, urine, cryptococcal DNA, HIV RNA and lymphocyte T CD4+ count), treatment and outcome aspects. We analyze 69 patients (71% men, 29% women) with HIV infection and cryptococcosis.&#x0D; Results: 69 cases of cryptococcosis were confirmed for 699 HIV infected hospitalized patients tested, giving a prevalence of 9.9%. 38% (n=26/69) of patients were with clinical signs of infection with the central nervous system involvement, 19% (13/69) patients had pulmonary involvement.&#x0D; Other 43% (n=30/69) of patients had disseminated non-CNS disease (elevated serum cryptococcal Ag or DNA). Most patients had advanced HIV disease (Median lymphocyte T CD4+ count=48, 5 cells/uL, (1-1041), the average was 112, 9 cells/ uL (SD 184.98). 87% (n=59/68) of patients had lymphocyte T CD4+cell count &lt; 200 cells/μL Only 25% (n=14) of the patients known to have HIV infection (n=56/69) were receiving antiretroviral therapy at the time of presentation. Overall mortality rate was 59% (n=41/69).&#x0D; Conclusion: Prevalence of cryptococcal antigenemia was 9.9%, indicating that the prevalence of cryptococcal infection among HIV patients in Latvia may be high enough to consider targeted screening.HIV positive patients have high mortality (35%) following cryptococcal infection which persists beyond their initial hospitalization. Follow-up studies of late mortality would be beneficial.

Selective and Sensitive Mass Spectrometric Identification of Immune Complex Antigens in Cerebrospinal Fluid.

Comprehensive identification of immune complex antigens (IC-antigens) in cerebrospinal fluid (CSF) is useful to provide insights into pathophysiology and could form the basis for novel diagnostic and treatment strategies for central nervous system autoimmune diseases and other neurological disorders. Immune complexome analysis is the method for comprehensively identifying IC-antigens in biological fluids (such as serum and CSF). Here, we describe IC-antigens detection method; specifically, ICs in CSF are captured and are subjected to papain-digestion elution and tryptic digestion, and are analyzed by nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS).

Occurrence of Leishmania infantum in the central nervous system of naturally infected dogs: Parasite load, viability, co-infections and histological alterations.

Zoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and little is known about the occurrence and pathogenesis of this parasite in the CNS. The aims of this study were to evaluate the occurrence, viability and load of L. infantum in the CNS, and to identify the neurological histological alterations associated with this protozoan and its co-infections in naturally infected dogs. Forty-eight Leishmania-seropositive dogs from which L. infantum was isolated after necropsy were examined. Cerebrospinal fluid (CSF) samples were analyzed by parasitological culture, quantitative real-time PCR (qPCR) and the rapid immunochromatographic Dual Path Platform test. Brain, spinal cord and spleen samples were submitted to parasitological culture, qPCR, and histological techniques. Additionally, anti-Toxoplasma gondii and anti-Ehrlichia canis antibodies in serum and distemper virus antigens in CSF were investigated. None of the dogs showed neurological signs. All dogs tested positive for L. infantum in the CNS. Viable forms of L. infantum were isolated from CSF, brain and spinal cord in 25% of the dogs. Anti-L. infantum antibodies were detected in CSF in 61% of 36 dogs. Inflammatory histological alterations were observed in the CNS of 31% of the animals; of these, 66% were seropositive for E. canis and/or T. gondii. Amastigote forms were associated with granulomatous non-suppurative encephalomyelitis in a dog without evidence of co-infections. The highest frequency of L. infantum DNA was observed in the brain (98%), followed by the spinal cord (96%), spleen (95%), and CSF (50%). The highest L. infantum load in CNS was found in the spinal cord. These results demonstrate that L. infantum can cross the blood-brain barrier, spread through CSF, and cause active infection in the entire CNS of dogs. Additionally, L. infantum can cause inflammation in the CNS that can lead to neurological signs with progression of the disease.

Clinical study on the cerebrospinal fluid antigen test for rapid diagnosis of pneumococcal meningitis

Objective To evaluate the value of Binax NOW pneumococcal antigen test for rapid diagnosis in patients with purulent meningitis. Methods Cerebrospinal fluid (CSF) were collected prospectively from 86 children who were diagnosed with purulent meningitis in the Department of Infectious Diseases at Beijing Children′s Hospital from March 2013 to March 2014 and were tested by Binax NOW method. CSF culture was considered as golden standard to evaluate the sensitivity and specificity of the Binax NOW method. Continuous variables were compared with t test, categorical variables were compared with χ2 test. Results Among 86 cases of purulent meningitis, 34 (39.5%) had positive culture and 21 (24.4%) were Streptococcus pneumonia, while 31 (36.0%) were positive by Binax NOW test. Using CSF culture as golden standard, the sensitivity and specificity of Binax NOW for diagnosis of pneumococcal meningitis were 90.5% (19/21) and 81.5% (53/65), respectively. Conclusion Binax NOW pneumoniae antigen test in CSF yields high sensitivity and specificity, which is useful in clinical application and enables prompt and adequate treatment. Key words: Purulent meningitis; Meningitis, Bacterial; Antigens; Streptococcus pneumoniae; Binax NOW

Significance of immune response to Mycobacterium tuberculosis culture filtrate protein antigens in cerebrospinal fluid of tuberculous meningitis patients: A search for diagnostic marker

ABSTRACTMycobacterium tuberculosis (H37Ra) culture filtrate proteins (CFP) are explored as a diagnostic marker for tuberculous meningitis (TBM). Cerebrospinal fluid (CSF) samples from patients were categorized as confirmed (n = 47), suspected (n = 20), and non-TBM (n = 25) cases. Immune response by Western blot revealed TBM CSF samples are having heterogeneous response to CFP. CFP ELISA was 92% sensitive and 38.30% specific. ODs of confirmed TBM and non-TBM cases were significantly different (P < 0.0001) and also the suspected TBM and non-TBM cases (P = 0.0001). No significant difference noticed in TBM and suspected TBM (P = 0.90). Thus, CFP can be a better biomarker for the diagnosis of TBM.

Cryptococcosis in Anti-Interferon-Gamma Autoantibody-Positive Patients: a Different Clinical Manifestation from HIV-Infected Patients

Disseminated nontuberculous mycobacterial (NTM) infection is the most common feature of patients who test positive for anti-interferon-gamma autoantibodies (IFN-γ Ab). Cryptococcus co-infects these patients. A retrospective, matched case-control study was conducted at Srinagarind Hospital between 1992 and 2013. We reviewed the medical records of non-HIV-infected patients with cryptococcosis and disseminated NTM infection (case) and matched 2 HIV-infected patients (control) with cryptococcosis to each case. There were 16 patients in the case group and 32 in the control group. Ten of the 16 patients in the case group had sera available for testing, and all tested positive for IFN-γ Ab. Compared to those in the control group, patients in the case group were significantly older, had a longer of illness duration and were less likely to present with meningitis but more likely to present with bone and/or joint, lung/pleura, and skin infections. Based on culture and staining results, patients in the case group were significantly less likely to test positive for the cryptococcal antigen in cerebrospinal fluid and serum samples, but were more likely to test positive in pus and skin lesion(s), compared to control patients. This is the first study of cryptococcosis in patients with disseminated NTM infection who tested positive for IFN-γ Ab. The clinical manifestations of cryptococcosis are different in these patients compared to those in HIV-infected patients. Recognizing the clinical features of these patients may improve diagnosis and promote timely treatment.

Diagnosis of Herpes Simplex Encephalitis by ELISA Using Antipeptide Antibodies Against Type-Common Epitopes of Glycoprotein B of Herpes Simplex Viruses

Herpes simplex encephalitis (HSE) represents one of the most severe infectious diseases of the central nervous system (CNS). As effective antiviral drugs are available, an early, rapid, and reliable diagnosis has become important. The objective of this article was to develop a sensitive ELISA protocol for herpes simplex viruses (HSV) antigen detection and quantitation by assessing the usefulness of antipeptide antibodies against potential peptides of HSV glycoprotein B (gB). A total of 180 cerebrospinal fluid (CSF) samples of HSE and non-HSE patients were analyzed using a panel of antipeptide antibodies against synthetic peptides of HSV glycoprotein gB. The cases of confirmed and suspected HSE showed 80% and 51% positivity for antipeptide against synthetic peptide QLHDLRF and 77% and 53% positivity for antipeptide against synthetic peptide MKALYPLTT, respectively for the detection of HSV antigen in CSF. The concentration of HSV antigen was found to be higher in confirmed HSE as compared to suspected HSE group and the viral load correlated well with antigen concentration obtained using the two antipeptides in CSF of confirmed HSE group. This is the first article describing the use of antibodies obtained against synthetic peptides derived from HSV in diagnostics of HSE using patients’ CSF samples.

Clinical and Laboratory Diagnosis of Central Nervous System Infections

Central nervous system (CNS) infections, including acute encephalitis, bacterial meningitis, tuberculous meningitis, and fungal meningitis, represent a potentially life-threatening neurological emergency. An etiological diagnosis of CNS infection is made a definite diagnosis based on the identification of pathogenic agents in the CNS using cerebrospinal fluid (CSF) smears or cultures, the identification of pathogenic antigens in CSF, the identification of pathogenic products in brain tissues or CSF with polymerase chain reaction (PCR), or the identification of specific antibody responses in CSF. Since time is required to obtain results from these laboratory tests, initial empirical treatments for the suspected CNS infection are required. The optimal specimen sampling and delivery is an essential requirement to reach the definite diagnosis. Nucleic acid amplification tests, such as PCR assays, show high sensitivity and specificity in detecting pathogens from CSF. In Japan, most genetic tests for pathogenic agents are performed as part of academic research. The number of commercially available genetic tests is limited. Moreover, no genetic tests for pathogenic agents in the CSF are covered by the national health insurance system in Japan. To manage the continuity of genetic tests, this inefficient system should be improved.

Identification of target antigens of naturally occurring autoantibodies in cerebrospinal fluid

Naturally occurring autoantibodies have natural physiologic functions related to normal cell processes. However, the repertoire of naturally occurring autoantibodies against neuronal antigens in CSF is unclear. The purpose of this study was to identify naturally occurring autoantibodies against neuronal antigens in CSF from patients with various neurologic diseases by proteomics-based analysis. The CSF samples were collected from 77 patients with various neurologic disorders. The antigen source for 2-dimensional immunoblotting was the SH-SY5Y human neuroblastoma cell line. There were 8 spots recognized in CSF from more than one-fourth of the 77 patients including all patient groups and these spots were recognized in intravenous immunoglobulin preparations. These antigen spots were identified as heat shock 105-kDa/110-kDa protein 1, isoform CRA_b, 78-kDa glucose-regulated protein, heat shock cognate 71-kDa protein, tubulin beta chain, vimentin (2 spots), and 60-kDa heat shock protein, mitochondrial; we could not identify the protein name corresponding to 1 of the 8 spots. In summary, there were 6 proteins identified that were main target antigens that reacted with naturally occurring autoantibodies in CSF from patients with varied neurologic disorders; the functions of autoantibodies against the identified antigens are unknown and may be clarified with further studies. Biological significanceNaturally occurring autoantibodies may have important functions in tissue homeostasis. In this study, we identified 6 common target antigens that reacted with autoantibodies in cerebrospinal fluid (CSF) from patients, independent of disease type. These findings may clarify the importance of naturally occurring autoantibodies in CSF and the use of these antibodies potentially may be a novel therapy for various neurologic disorders.

Cerebral aspergillosis and acute myeloid leukemia

A 32-year-old man with no previous medical history presented with weight-loss, frequent rhinorrhagia, gum swelling, cephalalgia, fatigue, fever, and somnolence. Neither motor nor sensory deficits were noted. The patient quickly became comatose and required mechanical ventilation. The complete blood count and the peripheral blood smear provided results compatible with leukemia (leukocytosis of 17,600/mm3, neutrophils 14,000/mm3, platelets 151,000/mm3, hemoglobin 13 g/dl, Auer rods). Consequently, bone marrow and CSF (cerebrospinal fluid) samples were collected and analyzed with the aid of cytochemistry and flow cytometry. The sandwich enzyme-linked immunosorbent assay (ELISA), “PlateliaAspergillus” (Bio-Rad Laboratories, Marnes La Coquette, France) detected galactomannan antigens both in CSF and serum. Brain magnetic resonance imaging [Figure ​[Figure1a1a-​-d]d] revealed a right frontal lesion with surrounding edema, which was ring enhancing after contrast administration. The DWI technique and the ADC values showed a limited diffusion [Figure ​[Figure2a2a and ​andb].b]. The diagnosis of cerebral aspergillosis was set. A stereotactic biopsy confirmed Aspergillus fumigatus as the cause of infection. Hyphae with branches of 45 degrees were seen (Grocott silver stain). The melanin-specific Masson-Fontana stain was used to exclude cerebral phaeohyphomycosis. Conidiophores of 8μm terminating in subclavate vesicles of 24 μm bearing uniseriate phialides were observed. The brain specimen was also inoculated on Sabouraud dextrose agar, and growth of white-dark grey colonies was observed. Amphotericin B (1 mg/Kg/day) was administered intravenously for a period of 6 weeks along with the liposomal formulation of daunorubicin (150 mg/m2) for 3 days. The new DWI images and the ADC map exhibited an increased diffusion [Figure ​[Figure2c2c and ​andd].d]. The primary focus of infection was not discovered. The patient failed to improve significantly, he did not regain consciousness and he eventually succumbed. An autopsy was not requested honoring the family's wishes. Figure 1 (a) T1: A hypo-intense lesion is depicted (right frontal lobe) (b) T2: A large area of edema is shown (c) FLAIR: The central portion exhibits a lower intensity signal as compared to the higher-signal perilesional area (angiogenic edema) (d) T1 postcontrast: ... Figure 2 (a) DWI (diffusion-weighted imaging) (b-value: 1,000): High signal of the lesion (diffusion is restricted) (b) ADC (apparent diffusion coefficient) values show a limited diffusion (values ranging between 0.45 and 0.65 mm2/s*10-3) (c) Same patient, 6 weeks ... This case represents one of a few reports depicting the presence of Aspergillus in the brain parenchyma in leukemia-harboring patients with the aid of modern magnetic resonance imaging techniques. Cerebral aspergillosis represents an opportunistic infection, which is commonly transmitted hematogenously.[1] It also manifests as a vascular insult or/and an abscess.[2] CT has always been the first line of evaluation but MRI still remains the method of choice due to its high sensitivity.[3,4] Additionally, diffusion techniques increase specificity,[1,2] and can evaluate the patients’ response to specific antifungal treatments. The three major neuroimaging findings in cerebral aspergillosis include infarction areas, ring lesions (post-infarct abscess formation), and dural/vascular infiltration.[5] Recognition of these patterns results in an effective treatment.[4,5] The authors’ scope was not only to draw the readership's attention to this rare fungal entity but also to raise awareness for the substantial help that new imaging modalities could provide in setting an accurate diagnosis for the benefit of the patient. Especially the absence or presence of restricted diffusion in the setting of brain fungal infections may be of great clinical utility.

Another Consequence of Severe Lupus: Invasive Fungal Disease

Infections are a substantial cause of morbidity and mortality in patients with systemic lupus erythematosus (SLE), accounting for one-quarter of all deaths1,2. The mechanisms for increased susceptibility to infections in patients with SLE are classically attributed to disease- and treatment-related factors. Polymorphisms in the mannose-binding lectin3 protein, which helps in host defense by activating the complement cascade, have been reported as a risk factor primarily for bacterial infections in SLE3. Patients with SLE may also have acquired hypogammaglobulinemia (as reviewed by Yong, et al 4) or hyposplenism, which leads to increased susceptibility to infections with encapsulated organisms. Acquired neutropenia has also been cited as a risk factor for infections in SLE5 and for fungal infections in other populations6. Corticosteroids are frequently necessary in the treatment of SLE, resulting in diminished cellular immunity. Immunosuppressant therapy also clearly increases the risk of bacterial infection7. Invasive fungal infections were initially defined in 20028, with revisions occurring in 2008, including a change in terminology to “invasive fungal disease” (IFD)9 by the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group. Proven IFD are subdivided into molds ( Aspergillus ) and yeasts ( Candida species and Cryptococcus ) and depend on culture or microscopic analysis from sterile anatomic sites, blood cultures, or cryptococcal antigen in cerebrospinal fluid. Separate criteria … Address correspondence to Dr. Barber; E-mail: cehbarbe{at}ucalgary.ca