Research On The Expression Of PD-1,Tim-3 And Cytokines In Children With Acute Dengue Virus Intracranial Infection

Objective To investigate the clinical characteristics of children with intracranial infection of acute dengue virus in Chenzhou Children’s Hospital,and detect the expression of Programed death-1(PD-1),T cell immuno-globulin and mucin domain 3(Tim-3)and related cytokines in peripheral blood and related cytokines in cerebrospinal fluid.Methods We collected blood and cerebrospinal fluid specimens from hospitalized children with acute viral encephalitis,meningitis and meningoencephalitis from May 2014 to September 2015 in Chenzhou Children’s Hospital.Dengue virus(DENV)were detected in CSF by the method of enzyme-linked immunosorbent assay(ELISA)and polymerase chain reaction(PCR)/reverse transcription-polymerase chain reaction(RT-PCR).Then we detected G-CSF,GM-CSF,IFN-γ,IL-10,IL-12(p70),IL-13,IL-17,IL-1b,IL-2,IL-4,IL-5,IL-6,IL-7,IL-8,MCP-1,TNF-αand MIP-1β in blood and CSF specimens from children with acute DENV intracranial infection by suspension chip technique.Finally,we detectedthe expression of PD-1 and Tim-3 on the surface of CD4+T cells,CD8+T cells and NK cells in children by flow cytometry.After referring to relevant cases and literature,combining with test results,we analyzed clinical features,PD-1,Tim-3 and cytokine expression levels of patients with acute DENV intracranial infection.Results1.A total of 245 patients with acute viral encephalitis,meningitis and meningoencephalitis were enrolled in this study.Twenty-six patients with DENV were detected by ELISA and RT-PCR.There were 25 patients with DENV-Ⅰ infection(96.2%),1 patient with DENV-Ⅱinfection(3.8%).2.Comparisons between plasma samples of patients with acute DENV intracranial infection and healthy control,the patients have higher G-CSF(p=0.018),IL-10(p=0.040)and lower IFN-γ(p=0.049)than controls.And other cytokines between the two groups have no significant difference.3.Comparisons between plasma and CSF samples of patients with acute DENV intracranial infection,the expression levels of IL-6(p=0.005),IL-8(p =0.004),G-CSF(p =0.000),GM-CSF(p=0.003)andMCP-1(p=0.000)are higher in CSF than in plasma.And other cytokines between the two groups have no significant difference.4.Comparisons between plasma samples of patients with acute DENV intracranial infection and healthy control,the patients have higher expression of PD-1,Tim-3 on the surface of CD4+T cells,CD8+T cells and NK cells than controls(p≤0.05).5.In plasma,the level of IFN-γ is negatively correlated with the level of IL-10(r=-0.440,p=0.040).6.The expression of IFN-γ in plasma of children with acute DENV intracranial infection is negatively correlated with the level of Tim-3expression in CD4+T cells,CD8+T cells and NK cells.(r=-0.651,p=0.001;r=-0.506,p=0.016;r=-0.423,p=0.045;respectively),The expression of IFN-γ in plasma of children with acute DENV intracranial infection is negatively correlated with the level of PD-1 expression in CD8+T cells and NK cells(r=-0.576,p=0.005;r=-0.475,p=0.026;respectively);while the expression of IL-10 in plasma of children with acute DENV intracranial infection is positively correlated with the level of Tim-3 expression in CD8+T cells and NK cells(r=0.475,p=0.021;r=0.492,p=0.011;respectively).Conclusions1.In children with acute DENV intracranial infection,the concentration of IL-10 and G-CSF in plasma increased,the concentration of IL-6,IL-8,G-CSF,GM-CSF,MCP-1 in cerebrospinal fluid increased,while the concentration of IFN-γ in plasma decreased,this may be related to its immune regulation mechanism.2.The levels of PD-1 and Tim-3 on CD4 + T cells,CD8 + T cells and NK cells in peripheral blood of children with acute DENV intracranial infection are significantly increased,the expression of PD-1and Tim-3 may be relevant to the expression of IFN-γ and IL-10.