Cerebrospinal Fluid Lymphocyte Subsets 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.

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.

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.

Immunological Markers of Optimal Response to Natalizumab in Multiple Sclerosis

To explore cell subsets and molecules that changed specifically in patients with multiple sclerosis (MS) who had an optimal response to natalizumab. Natalizumab is a monoclonal antibody that inhibits the migration of activated immune cells to the central nervous system. It shows high efficacy in modifying the natural history of MS and induces freedom of disease activity in about 40% of treated patients with MS. Prospective study of intrathecal immunoglobulin synthesis and cerebrospinal fluid lymphocyte subsets in patients with MS before and 1 year after beginning treatment with natalizumab. We monitored clinical and magnetic resonance imaging activity during a median time of 2 years. Two tertiary hospitals from the Spanish National Health Service. A total of 23 patients with MS. The differences between patients free of disease activity and patients with active disease during treatment. Of the 23 patients, 10 (43.5%) remained free of disease activity during follow-up. The remaining 13 patients (56.5%) had relapses or new lesions despite natalizumab therapy. We did not find differences in demographic variables or clinical data between both groups prior to natalizumab therapy. All patients showed a decrease in cerebrospinal fluid CD4(+) cells regardless of their response to treatment. Conversely, only patients free of disease activity showed a decrease in local IgM and, to a lesser extent, in IgG synthesis. They also showed lower percentages of B cells, particularly of CD5(+) and plasmablast subsets that virtually disappeared after treatment with natalizumab. These data indicate that inhibition of intrathecal antibody synthesis is associated with a complete therapeutic response to natalizumab in patients with aggressive MS.

CSF lymphocyte subsets in aseptic meningitis: dual-labelling analysis with flow cytometry

In order to characterize the CSF (cerebrospinal fluid) lymphocytes in CNS (central nervous system) inflammation, we examined paired samples of CSF and PB (peripheral blood) of 19 patients with acute aseptic meningitis, performing the dual labelling method on flow cytometry. Significantly higher percentages of CD3+ (T cell), CD4+ (helper-inducer), Leu3a+ Leu15- (cytotoxic-T) and HLA-DR+ CD3+ (activated-T) cells were identified in the CSF than in the PB of these patients. We observed significantly lower percentages of CD19+ (B cell), Leu2a+ Leu18+ (suppressor-inducer) and HLA-DR+ CD3- cells in the CSF than in the PB of these patients. Relative increases in helper-inducer, cytotoxic-T and activated-T cells in the CSF of aseptic meningitis are supposed to represent an active inflammatory process. However, whether these changes are specific or pathognomonic to any disease(s) remains to be solved.

Mononuclear cell subpopulations in CSF and blood of children with bacterial meningitis

To elucidate mononuclear cell subsets of the cerebrospinal fluid (CSF) in order to investigate the pathogenesis of bacterial meningitis (BM). Mononuclear cell and lymphocyte subsets in CSF and peripheral blood from 10 children with acute-stage BM before treatment on the same day were measured by flow cytometry. The control subjects for the subsets of peripheral blood were 15 healthy children. The percentages of CD14+ monocytes/macrophages (median: 56.5%), activated CD14+CD16+ monocytes/macrophages (20.9%), and CD14+CD16+ cells among total CD14+ cells (37.9%) in the CSF were significantly higher than those in the blood of children with BM (p<0.01, p<0.01, and p<0.05, respectively), which were significantly higher than those of the controls (p<0.001, p<0.001, p<0.05, respectively). The percentages of CD3+ (77.3%), CD4+ (45.2%), and CD8+ T cells (32.6%) in the CSF were significantly higher than those in the blood of affected children (p<0.01, p<0.01, and p<0.05, respectively). The percentages of CD3+, CD4+, and CD8+ T cells in the blood of children with BM were significantly lower than those of controls (all p<0.001). The percentages of CD20+ B cells (6.9%) in the CSF were significantly lower than those in the blood of affected children (p<0.01), which were significantly higher than those of controls (p<0.001). The percentages of monocytes/macrophages and T cells in CSF were higher than those in blood in children with BM.

Immunologic and Clinical Responses to Rituximab in a Child With Opsoclonus-Myoclonus Syndrome

Opsoclonus-myoclonus syndrome (OMS) is an autoimmune disorder with serious neurodevelopmental morbidity and limited treatment options. We treated a toddler with moderately severe OMS with rituximab, a monoclonal anti-B cell antibody. The patient's clinical response was documented on videotape and scored with the OMS Evaluation Scale. Cerebrospinal fluid lymphocyte subsets were evaluated by flow-cytometric immunophenotyping, with a comprehensive panel of monoclonal antibodies. Eradication of cerebrospinal fluid B cells, which previously were expanded, was associated with dramatic clinical improvement. There also were secondary changes in other lymphocyte subsets that might be relevant to the clinical response and lack of serious infections. In addition to clarifying the immune response to B-cell depletion, these data reveal a promising new therapy for OMS that warrants a phase I clinical trial.

Flow cytometric analysis of lymphocytes in cerebrospinal fluid in patients with tick-borne encephalitis.

Cerebrospinal fluid (CSF) lymphocyte subsets were examined by flow cytometry in 33 patients with tick-borne encephalitis (TBE) in order to determine their values. Lymphocytes were isolated from CSF and lymphocyte subsets were determined: lymphocytes T (CD3+), lymphocytes B (CD19+), NK cells (CD3-CD56+), helper T cells (CD3+CD4+) and cytotoxic T cells (CD3+CD8+). The expression of IL-2 receptors (CD25+) and transferrin receptors (CD71+) on T cells and HLA-DR molecules on T cell subsets was examined. Furthermore, possible relationships among different TBE patient population variables (gender, age, severity of disease, duration of meningitis) were considered. The analyses of the CSF lymphocyte population subsets are presented. Lymphocytes T (CD3+) were significantly higher in the CSF than in the peripheral blood as was the case with the T cells that expressed transferrin receptors (CD71). Lymphocytes B (CD19+) and NK cells (CD3-CD56+) prevailed in the peripheral blood. In the early course of the disease, a higher expression of HLA-DR molecules on T lymphocytes was observed, while later a higher expression of IL-2 receptors (CD25+) was observed. Significant differences in lymphocyte subsets between the CSF and the peripheral blood were found. Significant time-dependent changes of CSF lymphocyte subsets during course of infection were observed. The results of the present study give us deeper insight into CNS cellular immunopathogenic mechanisms in patients with TBE.

Cytoimmunological abnormalities in cerebrospinal fluid in early stages of HIV-1 infection often precede changes in blood

We determined the proportions of CD2 +, CD4 + and CD8 + lymphocytes in the cerebrospinal fluid (CSF) and paired blood samples of 19 neurologically examined subjects with CDC II/III stage HIV-1 infection and 21 controls. Using a three-layer indirect immunoperoxidase technique, we found that 16 HIV-1 infected subjects had an abnornally low CD4 +/CD8 + ratio of the CSF due to a decrease in the proportion of CD4 + T cells and/or an increase in that of CD8 + lymphocytes. Compared to controls, in the HIV-1-infected group this ratio appeared to be lower in both CSF and blood ( P < 0.001). The changes of lymphocyte subpopulations were more marked in the CSF than in blood, and no clear associations were found between the findings of paired CSF and blood samples. Cytological examination of the CSF revealed that 12 of 14 patients had an elevated total cell count and an abnormal differential count with a predominance of mononuclear phagocytes. An elevated number and/or proportion of enlarged lymphoid cells was found in most cases. The total cell counts, and total numbers of mononuclear phagocytes and lymphocytes of the patients were higher than in controls ( P < 0.001). Our study shows that the changes in CSF lymphocyte subsets often precede those in blood and that immunological events in the central nervous system (CNS)_are not directly reflected in peripheral blood. Cytoimmunological abnormalities in the CSF of individuals with early stages of HIV-1 infection indicate subclinical CNS involvement, and may be of importance for therapeutic considerations.

Quantitation of Lymphocyte Subsets in Cerebrospinal Fluid and Blood during the Clinical Course of Aseptic and Bacterial Meningitis

Mononuclear cell subsets in cerebrospinal fluid (CSF) and peripheral blood (PB) were monitored during the clinical course in 23 patients with acute meningitis using 6 monoclonal antibodies. Significant differences between aseptic and bacterial meningitis mainly consisted of a higher percentage of OKT4-positive cells in PB in the acute phase of bacterial meningitis. Significant differences between CSF and PB are found in the amount of most cell subtypes at all times except the acute phase of bacterial meningitis. The OKT4/OKT8 ratio was always significantly higher in CSF and correlated with the acuity of inflammation in bacterial meningitis.

Lymphocyte subsets at different stages of subacute sclerosing panencephalitis: a study with monoclonal antibodies.

Lymphocyte subsets in cerebrospinal fluid (CSF) and peripheral blood were studied using monoclonal antibodies, in patients with subacute sclerosing panencephalitis, eight of whom were at stage 2 and seven at stage 4. Eighteen subjects affected with non immunological diseases constituted the controls. Regardless of the stage, patients with subacute sclerosing panencephalitis had lower percentages of OKT3+ (pan-T) cells in both CSF and peripheral blood, with an increase of OKIa+ cells (B cells, macrophages and active T cells) in peripheral blood. A difference was found in the proportion of OKT4+ (helper-inducer) and OKT8+ (suppressor/cytotoxic) cells in relation to the stage, the most striking finding being a significant decrease of OKT8+ with an increase of T4/T8 ratio in peripheral blood at an early stage.