Cerebrospinal Fluid Lymphocytes Research Articles

High expression of CD3+T-lymphocytes in cerebrospinal fluid increases the risk of critical cerebral hemorrhage with systemic inflammatory response syndrome (SIRS) after surgery

ObjectiveTo assess the frequency of lymphocyte subsets and other laboratory indicators in paired cerebrospinal fluid (CSF) and peripheral blood (PB) samples from critically ill patients with intracerebral hemorrhage (ICH) who developed systemic inflammatory response syndrome (SIRS) following surgery. IntroductionNeuroinflammation and systemic inflammatory responses significantly contribute to secondary brain injury following ICH. Post-surgery SIRS is known to worsen clinical outcomes in ICH patients; however, the immune response in the CSF and PB has not been fully characterized. Understanding immunological changes in ICH patients with SIRS could lead to improved clinical management and prognostic outcomes. MethodsThis study involved a retrospective analysis of data from patients with ICH who underwent surgery in the Neurological Intensive Care Unit (NICU) of Baoding No. 1 Hospital, Hebei Province, China, between January and July 2022. Patients were divided into SIRS and non-SIRS groups based on the clinical criteria. Demographic, clinical, and laboratory data, including lymphocyte subsets in CSF and PB, were collected and analyzed. This study compared lymphocyte subsets and other inflammatory markers between the SIRS and non-SIRS groups. ResultsPatients with SIRS demonstrated higher systolic blood pressure (SBP) at admission, worse 90-day prognoses, elevated inflammatory markers, increased levels of complement proteins C3 and C4, and lower levels of immunoglobulin G (IgG) compared to patients without SIRS. Between 3–6 days post-surgery, SIRS patients showed higher percentages of CD3+T cells, CD4+T cells, and CD4+/CD8+ ratios in the CSF than non-SIRS patients. CD3+T cell percentages in the CSF were consistently higher than those in the PB and were independent of PB levels. In contrast, CD3-CD16+CD56+ natural killer (NK) cell percentages were lower in patients with SIRS. No significant differences in PB lymphocyte subsets were found between the two groups. A high CSF CD3+T cell percentage (≥85.68 %) was identified as the strongest predictor of critical ICH with SIRS after surgery, with an appropriate use criterion (AUC) of 0.7742, sensitivity of 77.42 %, specificity of 76.19 %, and 95 % CI of 0.6655–0.8829 (P < 0.0001). ConclusionElevated levels of CD3+T lymphocytes in CSF are strongly associated with an increased risk of severe cerebral hemorrhage and SIRS following surgery. These findings suggest that monitoring CSF immune markers, particularly CD3+T lymphocytes, could serve as valuable predictors for the development of SIRS in critically ill ICH patients and inform post-surgical treatment strategies.

Cognitive impairments in autoimmune encephalitis: the role of autoimmune antibodies and oligoclonal bands.

The presence of oligoclonal bands (OCBs) in cerebrospinal fluid (CSF) is a pivotal diagnostic marker for multiple sclerosis (MS). These bands play a crucial role in the diagnosis and understanding of a wide array of immune diseases. In this study, we explore the relationship between the cognitive profile of autoimmune encephalitis (AIE) and the presence of OCBs in CSF, with a particular emphasis on NMDA receptor antibodies. We studied a cohort of 21 patients across five tertiary centers, segregated into two distinct categories. One group comprised individuals who tested positive only for autoimmune encephalitis antibodies indicative of encephalitis, while the other group included patients whose CSF was positive for both autoimmune encephalitis antibodies and OCBs. Our investigation focused primarily on cognitive functions and behavioral alterations, supplemented by auxiliary diagnostic assessments such as CSF cell count, magnetic resonance imaging (MRI), and electroencephalogram (EEG) results, evaluated for the two patient groups. To validate our findings, we employed statistical analyses such as Fisher's exact test with Benjamini-Hochberg correction. Our study included 21 patients, comprising 14 who were presented with only autoimmune encephalitis antibodies, and 7 who were dual-positive. Among these patients, we focused on those with NMDA receptor antibodies. Of these, five were dual positive, and nine were positive only for NMDA receptor antibodies. The dual-positive NMDA group, with an average age of 27 ± 16.47 years, exhibited significantly higher CSF cell counts (p=0.0487) and more pronounced language and attention deficits (p= 0.0264). MRI and EEG results did not differ significantly between the groups. Our results point to OCBs as an additional marker of disease severity in AIE, especially in NMDA receptor-antibody positive patients, possibly indicating a broader inflammatory process, as reflected in elevated CSF lymphocytes. Regular testing for OCBs in cases of suspected AIE may aid in disease prognosis and identification of patients more prone to language and attention disorders, improving diagnosis and targeting treatment for these cognitive aspects.

Comparative study on the immune surveillance injury of blood cerebrospinal fluid barrier induced by exposure to lead acetate and nano-lead sulfide

Objective: To investigate the differences in terms of blood cerebrospinal fluid barrier immune surveillance injury by lead acetate and nano-lead sulfide exposure in order to provide basis for the study of their mechanism of nerve injury caused by exposure to lead and nano lead. Methods: In June 2015, forty-five SPF SD male rats were randomly divided into control group, lead acetate group (20 mg/kg) and nano-lead sulfide group (20 mg/kg), with 15 rats in each group. The rats were intragastric five times a week, for nine weeks. The numbers of CD4(+) T lymphocytes in blood and cerebrospinal fluid were detected by flow cytometry. The levels of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum and cerebrospinal fluid were detected by ELISA. The expressions and distribution of intercellular cell adhesion molecule-1 (ICAM-1) and CD4(+) T lymphocytes in choroid plexus were detected by laser confocal fluorescence immunoassay. The mRNA expression levels of IL-4, IFN-γ and ICAM-1 in the choroid plexus were detected by real-time PCR. Results: Compared with the control group, the proportion of CD4(+) T lymphocytes in blood of rats in lead acetate group was increased, the proportions of CD4(+) T lymphocytes in cerebrospinal fluid of rats in lead acetate group and nano-lead sulfide group were increased, the contents of IL-4 and IFN-γ in serum of rats in lead acetate group and nano-lead sulfide group were increased, the content of IL-4 in cerebrospinal fluid of rats in lead acetate group and the contents of IL-4 and IFN-γ in cerebrospinal fluid of rats in nano-lead sulfide group were increased, the differences were statistically significant (P<0.05). The fluorescence intensity of ICAM-1 and CD4(+) T lymphocytes in choriochoroid plexus of rats in lead acetate group and nano-lead sulfide group were stronger than those in control group, and the fluorescence intensity of CD4(+) T lymphocytes of rats in nano-lead sulfide group was weaker than that in lead acetate group. Compared with the control group, the mRNA expression levels of ICAM-1, IL-4 and IFN-γ in choriochoroids plexus of rats in lead acetate group and nano-lead sulfide group were increased, and the mRNA expression levels of ICAM-1 and IL-4 in nano-lead sulfide group were higher than those in lead acetate group, while the mRNA expression level of IFN-γ in nano-lead sulfide group was lower than that in lead acetate group (P<0.05) . Conclusion: Exposure to lead and nano-lead sulfide can cause the increase of CD4(+) T lymphocytes, IL-4, IFN-γ and ICAM-1, which may be related to the damage to the immune surveillance of the blood cerebrospinal fluid barrier. And there is a difference in the injury caused by lead and nano-lead sulfide exposure.

A comprehensive investigation into the ranges of laboratory tests present in cerebrospinal fluid across various types of meningitis within different age categories

Background. Diagnosing meningitis requires a lumbar puncture to analyze cerebrospinal fluid (CSF) and determine the causative pathogen. Key diagnostic measures include Glucose and Protein concentrations, cell count, differential leukocyte count, Gram stain, culture, and PCR if necessary. Objective. Our analysis assessed patterns in key biomarkers—neutrophils, lymphocytes, cerebrospinal fluid white blood cell count, and CSF/blood Glucose and Protein ratios—across various types of meningitis, including meningococcemia, meningococcal, tuberculous, pneumococcal, aseptic, and other bacterial meningitis. We examined these biomarkers within distinct age categories: children (0-12 years), adults (13-64 years), and the elderly (≥ 65 years). Methods. Descriptive statistics were used to analyze various types of meningitis across different age groups. We assessed the normality of laboratory biomarkers using the Shapiro-Wilk test, and applied Welch ANOVA to explore potential differences in biomarker levels among various types of meningitis. A Games-Howell post hoc test was used to identify specific pairs of meningitis types with statistically significant differences in biomarker levels. Results. The analysis of variance of cerebrospinal fluid biomarkers revealed significant differences in CSF WCC (cells/mm3), Neutrophils (%), Lymphocytes (%), CSF/blood Glucose and Protein ratios in children and adults (p &lt;0.05). However, differences were observed solely in neutrophil levels among elderly subjects. In bacterial meningitis, neutrophil levels increased across all ages, with adults showing higher levels. For meningococcal meningitis, adults had a median neutrophil level of 78.5% [60%, 90%] compared to 57% [24.5%, 85.25%] in children. The CSF white cell count (WCC) median was also higher in adults (339 cells/mm³) compared to children (224 cells/mm³). Viral meningitis cases exhibited higher lymphocyte levels across all age groups, with medians of 69% [34%, 87%] in children, 82% [61%, 93%] in adults, and 77% [55%, 90%] in the elderly. The glucose ratio was lower in bacterial meningitis (&lt;0.3) and higher in viral cases (&gt;0.5). Protein ratios were elevated in bacterial meningitis, indicating increased blood-brain barrier permeability. These results demonstrate a distinct biological profile for different causative agents, modulated by patient age.

A novel diagnostic model for tuberculous meningitis using Bayesian latent class analysis

BackgroundDiagnosis of tuberculous meningitis (TBM) is hampered by the lack of a gold standard. Current microbiological tests lack sensitivity and clinical diagnostic approaches are subjective. We therefore built a diagnostic model that can be used before microbiological test results are known.MethodsWe included 659 individuals aged ≥16\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\ge 16$$\\end{document} years with suspected brain infections from a prospective observational study conducted in Vietnam. We fitted a logistic regression diagnostic model for TBM status, with unknown values estimated via a latent class model on three mycobacterial tests: Ziehl–Neelsen smear, Mycobacterial culture, and GeneXpert. We additionally re-evaluated mycobacterial test performance, estimated individual mycobacillary burden, and quantified the reduction in TBM risk after confirmatory tests were negative. We also fitted a simplified model and developed a scoring table for early screening. All models were compared and validated internally.ResultsParticipants with HIV, miliary TB, long symptom duration, and high cerebrospinal fluid (CSF) lymphocyte count were more likely to have TBM. HIV and higher CSF protein were associated with higher mycobacillary burden. In the simplified model, HIV infection, clinical symptoms with long duration, and clinical or radiological evidence of extra-neural TB were associated with TBM At the cutpoints based on Youden’s Index, the sensitivity and specificity in diagnosing TBM for our full and simplified models were 86.0% and 79.0%, and 88.0% and 75.0% respectively.ConclusionOur diagnostic model shows reliable performance and can be developed as a decision assistant for clinicians to detect patients at high risk of TBM.SummaryDiagnosis of tuberculous meningitis is hampered by the lack of gold standard. We developed a diagnostic model using latent class analysis, combining confirmatory test results and risk factors. Models were accurate, well-calibrated, and can support both clinical practice and research.

Development and validation of a new model for the early diagnosis of tuberculous meningitis in adults based on simple clinical and laboratory parameters

BackgroundThe differential diagnosis between tuberculous meningitis (TBM) and viral meningitis (VM) or bacterial meningitis (BM) remains challenging in clinical practice, particularly in resource-limited settings. This study aimed to establish a diagnostic model that can accurately and early distinguish TBM from both VM and BM in adults based on simple clinical and laboratory parameters.MethodsPatients diagnosed with TBM or non-TBM (VM or BM) between January 2012 and October 2021 were retrospectively enrolled from the General Hospital (derivation cohort) and Branch Hospital (validation cohort) of Ningxia Medical University. Demographic characteristics, clinical symptoms, concomitant diseases, and cerebrospinal fluid (CSF) parameters were collated. Univariable logistic analysis was performed in the derivation cohort to identify significant variables (P < 0.05). A multivariable logistic regression model was constructed using these variables. We verified the performance including discrimination, calibration, and applicability of the model in both derivation and validation cohorts.ResultsA total of 222 patients (70 TBM and 152 non-TBM [75 BM and 77 VM]) and 100 patients (32 TBM and 68 non-TBM [31 BM and 37 VM]) were enrolled as derivation and validation cohorts, respectively. The multivariable logistic regression model showed that disturbance of consciousness for > 5 days, weight loss > 5% of the original weight within 6 months, CSF lymphocyte ratio > 50%, CSF glucose concentration < 2.2 mmol/L, and secondary cerebral infarction were independently correlated with the diagnosis of TBM (P < 0.05). The nomogram model showed excellent discrimination (area under the curve 0.959 vs. 0.962) and great calibration (P-value in the Hosmer–Lemeshow test 0.128 vs. 0.863) in both derivation and validation cohorts. Clinical decision curve analysis showed that the model had good applicability in clinical practice and may benefit the entire population.ConclusionsThis multivariable diagnostic model may help clinicians in the early discrimination of TBM from VM and BM in adults based on simple clinical and laboratory parameters.

Associations of the cerebrospinal fluid lymphocyte population with a clinical presentation of tick-borne encephalitis

In tick-borne encephalitis (TBE), lymphocytes infiltrating central nervous system are indispensable for the infection control, but also potentially immunopathogenic. To clarify their roles, we have evaluated cerebrospinal fluid (CSF) count of the main lymphocyte populations (considered as a proxy of the brain parenchyma lymphocytic infiltrate) in TBE patients and analyzed if they associate with clinical presentation, blood-brain barrier disruption and intrathecal antibody synthesis.We have studied CSF from 96 adults with TBE (50 with meningitis, 40 with meningoencephalitis, 6 with meningoencephalomyelitis), 17 children and adolescents with TBE and 27 adults with non-TBE lymphocytic meningitis. Th CD3+CD4+, Tc CD3+CD8+, double positive T CD3+CD4+CD8+, B CD19+ and NK CD16+/56+ cells were counted cytometrically with a commercial fluorochrome-stained monoclonal antibody set. The associations between the counts and fractions of these cells and clinical parameters were analyzed with non-parametric tests, p<0.05 considered significant.The TBE patients had lower pleocytosis with similar proportions of the lymphocyte populations compared to non-TBE meningitis. The different lymphocyte populations correlated positively with one another, as well as with CSF albumin, IgG and IgM quotients. The higher pleocytosis and expansion of Th, Tc and B cells associated with a more severe disease and neurologic involvement: Th with encephalopathy, myelitis and weakly with cerebellar syndrome, Tc with myelitis and weakly with encephalopathy, B with myelitis and with at least moderately severe encephalopathy. The double-positive T lymphocytes associated with myelitis, but not with other forms of CNS involvement. The fraction of double positive T cells decreased in encephalopathy and the fraction of NK in patients with neurologic deficits. In children with TBE, Tc and B counts were increased at the expense of Th lymphocytes in comparison with adults.The concerted intrathecal immune response, involving the main lymphocyte populations, increases with the clinical severity of TBE, with no evidently protective or pathogenic elements distinguishable. However, the particular populations including B, Th and Tc cells associate with different, though overlapping, spectra of CNS manifestations, suggesting they may be specifically related to TBE manifesting as myelitis, encephalopathy and cerebellitis. The double-positive T and NK cells do not expand evidently with severity and may be most closely associated with the protective anti-TBEV response.

Neurological Manifestations of Scrub Typhus in Children: Clinical Profile and Outcome

Abstract Background: Re-emerging scrub typhus is gaining recognition as an important cause of focal or disseminated vasculitis or perivasculitis which is responsible for its various clinical manifestations and complications. Neurological manifestations with varied presentations have been reported in rickettsial diseases with variable prevalence (28%–80%). Methods: A cross-sectional study among admitted children was conducted for 1½ years in a tertiary care hospital with the objective to study the clinical profile and outcome of the patients with neurological manifestations of scrub typhus. Results: Out of 105 scrub typhus cases (immunoglobulin M enzyme-linked immunosorbent assay positive), 32 patients (30.4%) had neurological manifestations with a male: female ratio of 19:13. These children presented with fever (100%), headache (71.8%), vomiting (62.5%), convulsion (53.1%), and altered sensorium (43.7%). Neurological manifestations included meningeal signs (75%), ataxia (6.2%), lateral rectus palsy (9.3%), papilledema (18.7%), stroke (6.2%), and retinal hemorrhage (3.1%). Lumbar puncture was done in 31 cases and the mean cerebrospinal fluid (CSF) white blood cell count, lymphocyte, and neutrophil proportion was 66 cells/cu.mm, 60%, and 27%, respectively, with the mean CSF protein and glucose is 67 mg/dl and 55 mg/dl. Neuroimaging was done in 24 patients and the abnormalities found were cerebral edema (n = 5), basal ganglia infarct (n = 2), and features of cerebellitis (n = 2). Conclusion: Rickettsial disease should be considered an etiological agent in all febrile illnesses with neurological manifestations, especially in endemic areas. Early recognition and treatment of rickettsial infections with neurological manifestations are important to prevent morbidity and mortality of the disease.

Association of cerebrospinal fluid CD4+/CD8+Ratio with 60-day functional outcome after intracerebral hemorrhage.

Background: The immune inflammatory reaction has vital function in pathologic mechanism of critical intracerebral hemorrhage. It recently has been reported that CD4/CD8 ratio may represent a novel composite immune inflammatory marker to predict prognosis of critical intracerebral hemorrhage (ICH). Nevertheless, as for considering the effects of surgical evacuation upon initiation of immune inflammatory reactions, the association between cerebrospinal fluid (CSF) CD4/CD8 ratio and 60-day functional outcome of patients with critical ICH after surgery has not been investigated. Present study aimed to evaluate the predictive value concerning postoperative complement system and immunoglobulin, paired cerebrospinal fluid and peripheral blood lymphocyte subsets, as well as inflammation index before and after the operations upon the 60-day prognosis of patients with ICH.Methods: In total, 69 patients with acute critical ICH admitted in First Central Hospital of Baoding City from January to July in 2022 were prospectively enrolled. We recorded and analyzed the relevant clinical data. Laboratory parameters included postoperative lymphocyte subsets in paired cerebrospinal fluid and peripheral blood, inflammation index before and after operation. The associations between 60-day outcome and laboratory biomarkers were assessed by multivariable logistic regression analysis. Comparisons of predictive value regarding independent predictors was evaluated by receiver operating characteristic (ROC) curves.Results: In total, 51 patients with critical ICH exhibited poor outcomes at 60days, which was associated with fever after surgery, hernia before surgery, SAH and lower Glasgow Coma Scale (GCS) at admission and large hematoma volume, greater CD3T%CSF, greater CD4T%CSF, and greater CD4/CD8 ratioCSF. CD4/CD8ratio CSF showcased significant predictive power by comparing with other laboratorial variables (AUC = 0.6808; cut-off = 1.61; sensitivity = 80.39%; specificity = 61.11%; 95% CI: 0.5232-0.8385; p = .0233), which was found to correlated linearly with postoperative fever, first CSF test time, CD3T% CSF, CD4T% CSF, CD8T% CSF, NKCSF, CD3T%PB, CD8T%PB, CD4/CD8 ratioPB, and glucoseCSF. Poor outcome at 60days linearly correlated with CD4/CD8ratioCSF after adjustments. In 3-5days after surgery tested CSF lymphocyte subsets, CD4/CD8ratioCSF ≥1.61 was associated with a higher risk for 60-day poor outcome comparing with corresponding subgroups.Conclusions: In association of critical ICH patient prognosis, CSF CD4/CD8 ratio, especially in 3-5days after surgery, exhibited potential independent predictive ability for 60-day functional outcomes of patients with critical ICH.

Investigation of white blood cell characteristics in cerebrospinal fluid samples at pediatric brain tumor diagnosis.

The role of white blood cells (WBC) in the pediatric central nervous system (CNS) tumor microenvironment is incompletely defined. We hypothesized that the WBC profile in cerebrospinal fluid (CSF) correlates with the presence of tumor cells and prognosis in pediatric CNS tumors, as well as other patient and disease characteristics, and differs by tumor type, thus giving insight into the tumor immune response. We conducted a retrospective analysis of CSF WBC profiles at CNS tumor diagnosis in 269 patients at our institution. We examined total nucleated cell count, absolute counts, and percentages by WBC subtype. We compared CSF WBC values by tumor cell presence, patient vital status, tumor location, and the most common tumor types. Patients who died of their tumor had a lower CSF lymphocyte percentage and a higher absolute monocyte count in CSF at diagnosis. The presence of tumor cells in CSF was associated with fewer lymphocytes and monocytes. Ventricular tumors had higher CSF lymphocyte, monocyte, macrophage, and total nucleated cell counts than extraventricular tumors. Germ cell tumors, low-grade glioma, high-grade glioma, and ependymoma had lower macrophage counts or percentages compared to other tumor types. WBC profile in CSF at pediatric CNS tumor diagnosis correlates with patient prognosis and presence of metastatic cells, along with tumor type and other tumor characteristics like relationship to the ventricles. Prospective CSF profiling and study may be useful to future immunotherapy and other pediatric CNS tumor clinical trials.

Role of SPAK\u2013NKCC1 signaling cascade in the choroid plexus blood\u2013CSF barrier damage after stroke

BackgroundThe mechanisms underlying dysfunction of choroid plexus (ChP) blood–cerebrospinal fluid (CSF) barrier and lymphocyte invasion in neuroinflammatory responses to stroke are not well understood. In this study, we investigated whether stroke damaged the blood–CSF barrier integrity due to dysregulation of major ChP ion transport system, Na+–K+–Cl− cotransporter 1 (NKCC1), and regulatory Ste20-related proline-alanine-rich kinase (SPAK).MethodsSham or ischemic stroke was induced in C57Bl/6J mice. Changes on the SPAK–NKCC1 complex and tight junction proteins (TJs) in the ChP were quantified by immunofluorescence staining and immunoblotting. Immune cell infiltration in the ChP was assessed by flow cytometry and immunostaining. Cultured ChP epithelium cells (CPECs) and cortical neurons were used to evaluate H2O2-mediated oxidative stress in stimulating the SPAK–NKCC1 complex and cellular damage. In vivo or in vitro pharmacological blockade of the ChP SPAK–NKCC1 cascade with SPAK inhibitor ZT-1a or NKCC1 inhibitor bumetanide were examined.ResultsIschemic stroke stimulated activation of the CPECs apical membrane SPAK–NKCC1 complex, NF-κB, and MMP9, which was associated with loss of the blood–CSF barrier integrity and increased immune cell infiltration into the ChP. Oxidative stress directly activated the SPAK–NKCC1 pathway and resulted in apoptosis, neurodegeneration, and NKCC1-mediated ion influx. Pharmacological blockade of the SPAK–NKCC1 pathway protected the ChP barrier integrity, attenuated ChP immune cell infiltration or neuronal death.ConclusionStroke-induced pathological stimulation of the SPAK–NKCC1 cascade caused CPECs damage and disruption of TJs at the blood–CSF barrier. The ChP SPAK–NKCC1 complex emerged as a therapeutic target for attenuating ChP dysfunction and lymphocyte invasion after stroke.

Main lymphocyte subpopulations in cerebrospinal fluid and peripheral blood in HIV-1 subtypes C and B.

HIV-1 subtype C (HIV-1C) shows reduced Tat protein chemoattractant activity compared with HIV-1B. The impact of HIV-1C Tat on the chemotaxis of the main lymphocyte subpopulations in the cerebrospinal fluid (CSF) and the peripheral blood (PB) is unclear. We hypothesized that there would be a lower frequency of specific lymphocyte subpopulations CD3+ or CD19+ in CSF in HIV-1C than in HIV-1B. The objectives were to detect the differences in the proportions of main lymphocyte subpopulations in CSF and PB, between people with HIV (PWH) and HIV-1-uninfected volunteers (PWoH) and in HIV-1B and HIV-1C. Lymphocyte immunophenotyping was studied in CSF and paired PB samples of PWH (n = 22) and PWoH (n = 14). Lymphocytes were analyzed within the CD45+ gated region. The proportions of CSF CD3+CD4+, CD3+CD8+, and CD3-CD19+ lymphocytes in CSF were comparable in HIV-1B and C. There was an increase in the proportion of CD3+CD8+ cells and a decrease in CD3+CD4+ T cells (ps = 0.016) in the CSF samples of the PWH compared with the PWoH group. In the PWH group, both CD3+CD4+ and CD3+CD8+ lymphocytes were significantly higher in the CSF than in the PB (p = 0.047 and 0.005). The proportion of CD3+CD4+ was lower and that of CD3+CD8+ was higher in the CSF samples of the aviremic group than that of HIV-negative control (p = 0.0008 and < 0.0001, respectively). HIV-1C Tat substitution (C30S) did not interfere with the CNS migration of the main lymphocyte subpopulations. This is the first study to evaluate these lymphocytes in CSF and PB of HIV-1C compared with HIV-1B.

Primary central nervous system lymphoma - an overview

Introduction. Primary central nervous system lymphoma is a rare entity mostly presenting with non-GCB diffuse large B-cell lymphoma, being confined to the brain, spinal cord, meninges, and eyes. Diagnosis. The diagnosis is frequently established by stereotactic or open the brain biopsy, but in some cases with isolated leptomeningeal involvement, the only way is to identify atypical/monoclonal lymphocytes in cerebrospinal fluid. By workup, we aim to define the extent of disease in the central nervous system and to exclude systemic involvement. Treatment. Treatment is tailored according to the patient?s age, fitness, vital organ function, comorbidities, and available therapy. The backbone of induction treatment is high-dose methotrexate, usually within polychemotherapy. Consolidation phase is a matter of debate between two approaches: 1. high dose chemotherapy with autologous stem cell transplantation, which appears to be the preferable option for young fit patients, and 2. whole brain radiotherapy, preserved for transplant-ineligible ones. Whole brain radiotherapy has been raising concerns because of frequent cognitive impairment, which has been significantly diminished by reducing the irradiation dose. Despite a comprehensive treatment approach, many patients relapse, and since the prognosis of relapsed/refractory disease is devastating, there is a sense of urgency for novel treatment strategies. Several targeted agents and immunomodulatory drugs have been investigated in the settings of both relapsed/refractory and initial therapy, but with limited success. Ibrutinib monotherapy can induce durable remissions in the first line, but in relapse/refractory settings, the results are controversial. Conclusion. Adequate patient selection and new prospective trials should improve survival and preserve the patient?s neurological status.

Diagnostic Model for Discrimination Between Tuberculous Meningitis and Bacterial Meningitis.

BackgroundThe differential diagnosis between tuberculous meningitis (TBM) and bacterial meningitis (BM) remains challenging in clinical practice. This study aimed to establish a diagnostic model that could accurately distinguish TBM from BM.MethodsPatients with TBM or BM were recruited between January 2017 and January 2021 at Tongji Hospital (Qiaokou cohort) and Sino-French New City Hospital (Caidian cohort). The detection for indicators involved in cerebrospinal fluid (CSF) and T-SPOT assay were performed simultaneously. Multivariate logistic regression was used to create a diagnostic model.ResultsA total of 174 patients (76 TBM and 98 BM) and another 105 cases (39 TBM and 66 BM) were enrolled from Qiaokou cohort and Caidian cohort, respectively. Significantly higher level of CSF lymphocyte proportion while significantly lower levels of CSF chlorine, nucleated cell count, and neutrophil proportion were observed in TBM group when comparing with those in BM group. However, receiver operating characteristic (ROC) curve analysis showed that the areas under the ROC curve (AUCs) produced by these indicators were all under 0.8. Meanwhile, tuberculosis-specific antigen/phytohemagglutinin (TBAg/PHA) ratio yielded an AUC of 0.889 (95% CI, 0.840–0.938) in distinguishing TBM from BM, with a sensitivity of 68.42% (95% CI, 57.30%–77.77%) and a specificity of 92.86% (95% CI, 85.98%–96.50%) when a cutoff value of 0.163 was used. Consequently, we successfully established a diagnostic model based on the combination of TBAg/PHA ratio, CSF chlorine, CSF nucleated cell count, and CSF lymphocyte proportion for discrimination between TBM and BM. The established model showed good performance in differentiating TBM from BM (AUC: 0.949; 95% CI, 0.921–0.978), with 81.58% (95% CI, 71.42%–88.70%) sensitivity and 91.84% (95% CI, 84.71%–95.81%) specificity. The performance of the diagnostic model obtained in Qiaokou cohort was further validated in Caidian cohort. The diagnostic model in Caidian cohort produced an AUC of 0.923 (95% CI, 0.867–0.980) with 79.49% (95% CI, 64.47%–89.22%) sensitivity and 90.91% (95% CI, 81.55%–95.77%) specificity.ConclusionsThe diagnostic model established based on the combination of four indicators had excellent utility in the discrimination between TBM and BM.

Cryptococcosis in patients with liver cirrhosis: Death risk factors and predictive value of prognostic models

BackgroundLiver cirrhosis is associated with immune deficiency, which causes these patients to be susceptible to various infections, including cryptococcus infection. Mortality in cirrhotic patients with cryptococcosis has increased. The present study was to explore the risk factors of mortality and the predictive ability of different prognostic models. MethodsForty-seven cirrhotic patients with cryptococcosis at a tertiary care hospital were included in this retrospective study. Data on demographics, clinical parameters, laboratory exams, diagnostic methods, medication during hospitalization, severity scores and prognosis were collected and analyzed. Student's t test and Mann-Whitney test were used to compare characteristics of survivors and non-survivors at a 90-day follow-up and cerebrospinal fluid (CSF) manifestations of cryptococcal meningitis. Multivariate Cox regression analysis was used to identify the independent risk factors for mortality. Kaplan-Meier curves were used to analyze patient survival. Receiver operating characteristic (ROC) curves were used to evaluate the different prognostic factors. ResultsThe 30- and 90-day survival rates were 93.6% and 80.9%, respectively, in cirrhotic patients with cryptococcosis. Cryptogenic liver diseases [hazard ratio (HR) = 7.567, 95% confidence interval (CI): 1.616-35.428, P = 0.010], activated partial thromboplastin time (APTT) (HR = 1.117, 95% CI: 1.016-1.229, P = 0.022) and Child-Pugh score (HR = 2.146, 95% CI: 1.314-3.504, P = 0.002) were risk factors for 90-day mortality in cirrhotic patients with cryptococcosis. Platelet count (HR = 0.965, 95% CI: 0.940-0.991, P = 0.008) was a protective factor. APTT (HR = 1.120, 95% CI: 1.044-1.202, P = 0.002) and Child-Pugh score (HR = 1.637, 95% CI: 1.086-2.469, P = 0.019) were risk factors for 90-day mortality in cirrhotic patients with cryptococcal meningitis. There was significant difference in the percentage of lymphocytes in CSF between survivors and non-survivors [60.0 (35.0-75.0) vs. 95.0 (83.8-97.2), P < 0.001]. The model of end-stage liver disease-sodium (MELD-Na) score was more accurate for predicting 30-day mortality both in patients with cryptococcosis [area under curve (AUC): 0.826, 95% CI: 0.618-1.000] and those with cryptococcal meningitis (AUC: 0.742, 95% CI: 0.560-0.924); Child-Pugh score was more useful for predicting 90-day mortality in patients with cryptococcosis (AUC: 0.823, 95% CI: 0.646-1.000) and those with cryptococcal meningitis (AUC: 0.815, 95% CI: 0.670-0.960). ConclusionsThese results showed that cryptogenic liver diseases, APTT and Child-Pugh score were associated with mortality in cirrhotic patients with cryptococcosis and cryptococcal meningitis. MELD-Na score was important for predicting 30-day mortality, and Child-Pugh score was critical for predicting 90-day mortality.

Artificial Intelligence in Differential Diagnostics of Meningitis: A Nationwide Study.

Differential diagnosis between bacterial and viral meningitis is crucial. In our study, to differentiate bacterial vs. viral meningitis, three machine learning (ML) algorithms (multiple logistic regression (MLR), random forest (RF), and naïve-Bayes (NB)) were applied for the two age groups (0–14 and >14 years) of patients with meningitis by both conventional (culture) and molecular (PCR) methods. Cerebrospinal fluid (CSF) neutrophils, CSF lymphocytes, neutrophil-to-lymphocyte ratio (NLR), blood albumin, blood C-reactive protein (CRP), glucose, blood soluble urokinase-type plasminogen activator receptor (suPAR), and CSF lymphocytes-to-blood CRP ratio (LCR) were used as predictors for the ML algorithms. The performance of the ML algorithms was evaluated through a cross-validation procedure, and optimal predictions of the type of meningitis were above 95% for viral and 78% for bacterial meningitis. Overall, MLR and RF yielded the best performance when using CSF neutrophils, CSF lymphocytes, NLR, albumin, glucose, gender, and CRP. Also, our results reconfirm the high diagnostic accuracy of NLR in the differential diagnosis between bacterial and viral meningitis.

The chemotactic cytokines in the cerebrospinal fluid of patients with neuroborreliosis

BackgroundThe outcome of neuroborreliosis (NB) is variable and may partially depend on host-related immune factors. In NB, the cerebrospinal fluid (CSF) contains a large population of T lymphocytes, but the mechanisms and consequences of their recruitment have not been fully elucidated. We have studied expression of T lymphocyte chemoattractant cytokines in association with CSF cytometric parameters and clinical data in NB patients. MethodsThe blood and CSF of 17 patients with NB and blood of 12 patients with erythema migrans (EM) were obtained before the antibiotic administration, and in fraction of NB patients during and/or after antibiotic treatment. The control samples came from blood donors (blood) and patients in whom neuroinfection was excluded by a lumbar puncture (CSF). Concentrations of IL-16, CXCL9, CXCL10, CXCL11, CCL2 and CCL5 in serum and CSF were measured with commercial ELISA. Data were analyzed with non-parametric tests, p < 0.05 considered significant. ResultsThe serum concentrations of IL-16, CXCL9, CXCL10 and CCL5 were increased, higher in NB than in EM. In CSF all the cytokines were upregulated, CXCL10, CXCL9 and IL-16 over ten-fold. The CSF concentration index favored the intrathecal synthesis of all the cytokines except CCL5, for which it could not be reliably estimated. CCL2, CXCL10 and CXCL9 created concentration gradients towards CSF. The intrathecal expression of IL-16, CCL5 and CXCL9 correlated with CSF lymphocyte counts, of IL-16, CXCL9 and CXCL10 - with a blood-brain barrier disruption, and of CXCL9 and CXCL10 with intrathecal specific IgG synthesis. The expression of CCL2, CXCL10 and CXCL11 peaked early after NB onset and decreased naturally afterwards. High initial CSF CXCL9, CXCL10 and CXCL11 levels associated with a persistent CSF pleocytosis and BBB disruption after treatment, but no cytokine was predictive of clinical outcome. In follow up (post-treatment) examinations, CSF CXCL10 and CCL5 associated positively and CCL2 negatively with a protracted lymphocytic pleocytosis. ConclusionsSeveral cytokines chemotactic for T lymphocytes are upregulated intrathecally in NB, with different dynamics and relation to other inflammatory parameters, suggesting their distinct pathogenetic roles. CXCL10 and CXCL9 are vividly upregulated and seem deeply involved in the pathogenesis of the intrathecal inflammation. IL-16 and CCL5 may directly drive T lymphocyte migration from periphery, but their ability to create an adequate chemotactic gradient remains to be confirmed. A delayed normalization of pleocytosis is accompanied by higher intrathecal expression of Th1-related and lower of Th2-related chemokines, in agreement with the protective role of Th1 to Th2 transition in the course of NB.

Major and minor lymphocytes subpopulations in peripheral blood and cerebrospinal fluid of children with meningitis

Introduction. The analysis of current publications indicates at our insufficient understanding of subpopulation composition of lymphocytes in peripheral blood and cerebrospinal fluid (CSF) during pediatric neuroinfectious diseases. It has been found that the main lymphocyte populations are divided into many small (minor) subpopulations.The purpose of this research was to assess percentage of major and minor blood and CSF lymphocyte subsets in children with aseptic viral meningitis (AM) or bacterial purulent meningitis (BM).Materials and methods. Phenotyping of blood and CSF lymphocytes of children aged from 4 months to 17 years diagnosed with AM (n = 86) and BM (n = 39) was carried out by using flow cytometry. As a comparison group, we analyzed peripheral blood and CSF samples collected from children with acute respiratory viral infections (ARVIs) associated with syndrome of meningism (n = 27). There was evaluated percentage of the major cell subpopulations (CD3+ T-lymphocytes, T-helpers — CD3+CD4+ Th, cytotoxic T-lymphocytes — CD3+CD8+ CTL, natural killer cells — CD3-CD16+CD56+ NK, B-cells — CD3-CD19+), as well as minor lymphocyte subsets (double positive (DP) (CD3+CD4+CD8+), double negative (DN) (CD3+CD4-CD8-) T-cells, NKT (CD3+CD16+CD56+), CD3-CD8+ NK, CD3+CD8dim and CD3+CD8 8bright).Results. It was found that the acute period of BM and AM vs. the comparison group (ARVI) was characterized by significant differences in the blood and CSF composition of major and minor lymphocyte subsets. In particular, blood T-cells, Th, CTL, NK, NKT, DN, CD3-CD8+ NK, CD3+CD8bright and CD3+CD8dim dominated in parallel with significantly lowered B-cell frequency in AM vs. BM. In the CSF of children with AM, T-cells and Th prevailed, whereas count of B-cells and CD3-CD8+ NK was lower compared to those in BM. In addition, further differences were revealed in CSF and blood cell subset composition depending on nosological entity, while maintaining differences in some major and minor lymphocyte subpopulations lacked in the comparison group. Calculating the CSF/blood ratio for the major and minor lymphocyte subsets uncovered the prevalence for the majority of cell subpopulations (the coefficients ranged from 1.2 to 16.4) in the CSF of the comparison group (ARVI), except B-cells, NK and CD3-CD8+ NK (coefficients ranged from 0.07 to 0.31). AM and BM were featured with various changes in the CSF/blood ratio found for most of the studied subpopulations in the acute period as well as the recovery phase highlighted with characteristic traits for each nosological form.Conclusion. The data obtained indicate about finding specific features in the activation of systemic and intrathecal immune response during viral and bacterial meningitis in children, which may be used as an additional differential diagnostic criterion.

The lymphocyte populations and their migration into the central nervous system in tick-borne encephalitis

In tick-borne encephalitis (TBE) the cerebrospinal fluid (CSF) cytosis is dominated by T CD3+CD4+ and T CD3+CD8+ lymphocytes, but their pathogenetic roles and mechanisms of migration into central nervous system (CNS) are unclear. Currently, we have studied CSF lymphocyte subsets and chemotactic axes in TBE patients stratified according to the clinical presentation. Blood and CSF were obtained from 51 patients with TBE (presenting as meningitis in 30, meningoencephalitis in 18 and meningoencephalomyelitis in 3), 20 with non-TBE meningitis and 11 healthy controls. We have studied: (1) abundances of the main lymphocyte subsets and (2) CXCR3 and CCR5 expression on CD3+CD4+ and CD3+CD8+ lymphocytes cytometrically with fluorochrome-stained monoclonal antibodies; (3) concentrations of chemotactic cytokines: CCL5 (CCR5 ligand), CXCL10 (CXCR3 ligand), IL-16, CCL2, CCL20 and CXCL5 with ELISA. Cytokine concentrations were additionally studied in 8 pediatric TBE patients. Data were analyzed with non-parametric tests, p < 0.05 considered significant. The higher CSF lymphocyte counts were associated with symptoms of CNS involvement, especially with altered consciousness (B, Th and Tc cells) and focal neurologic deficits (B cells). The minor fraction of double-positive T CD4+CD8+ cells was unique in associating negatively with encephalitis and altered consciousness. CSF CD3+CD4+ and CD3+CD8+ lymphocyte population was enriched in CCR5-positive cells and CCL5 concentration in CSF was increased and associated with a milder presentation. Although CXCL10 was vividly up-regulated intrathecally and correlated with CSF T lymphocyte counts, the CXCR3 expression in CSF T lymphocytes was low. Serum and CSF concentrations of CCL2, CXCL5 and IL-16 were increased in adult TBE patients, CCL2 created a chemotactic gradient towards CSF and both CCL2 and IL-16 concentrations correlated positively with CSF lymphocyte counts. The particular lymphoid cell populations in CSF associate differently with the clinical presentation of TBE, suggesting their distinct roles in pathogenesis. CCR5/CCL5 axis probably contributes to T lymphocyte migration into CNS. CXCL10 mediates the intrathecal immune response, but is probably not directly responsible for T cell migration. Additional chemotactic factors must be involved, probably including CCL2 and IL-16.

Evaluation of cerebrospinal fluid CXCL13 concentrations and lymphocyte subsets in tick-borne encephalitis

ObjectivesRecent studies suggest that the clinical presentation of tick-borne encephalitis (TBE) is determined by the host immune responses to the tick-borne encephalitis virus (TBEV). The aim of the study was to characterize immune responses in TBE to give a better insight into the immunopathogenesis of this disease. MethodsAnti-TBEV antibody levels, cerebrospinal fluid (CSF) and blood lymphoid populations, and concentrations of CXCL13 (a potent B-cell and T-cell chemoattractant), were analyzed in 35 patients with TBE (20 adults and 15 children). ResultsWhen compared with the blood, the CSF lymphoid population was significantly enriched in CD4+ T-cells and relatively depleted in natural killer (NK) cells and B lymphocytes. In comparison with TBE meningitis, patients suffering from TBE meningoencephalitis (n = 11, 31%) had a 3.5-fold higher median CSF CXCL13 concentration, 1.8-fold higher CSF/serum ratio of anti-TBEV IgG antibodies, and 1.8-fold higher median CSF cell count. CSF CXCL13 levels did not change significantly in children with TBE meningitis receiving supportive treatment, but decreased in children with TBE meningoencephalitis who received intravenous steroids. ConclusionsCD4+ cells are abundant in the CSF of patients with TBE. CXCL13 may be involved in the neuropathology of TBE by attracting different subsets of lymphocytes to the CSF.