The Expression Of Th1-Th2Cytokines In The Cerebrospinal Fluid Of Patients With Cryptococcus Meningitis

Objective: Cryptococcus meningitis (CM) is the most common fungalinfection of the central nervous system, caused by Cryptococcus neoformansinfection of the meninges. At present, scientists discover the occurrence,development and prognosis of Cryptococcus meningitis some extent dependon the patient’s own immune function and the imbalance of helper Tlymphocyte (Th1/Th2). In this study, we use cerebrospinal fluid cytology todynamic monitor the disease outcome status of patients with Cryptococcusmeningitis. We also detect the levels of Th1cytokine IFN-γ, TNF-α and Th2cytokines IL-10proteins in the CSF by ELISA to explore the hidden role ofthe local Th1/Th2immune response in the disease pathophysiology process ofCryptococcus meningitis.Methods:25Cryptococcus meningitis patients who received therapy inthe Second Hospital of Hebei Medical University between June2009and May2012were selected. All the25patients were confirmed by cerebrospinal fluidpathogen detection, use CSF MGG joint Allie new staining. Patients weredivided into the acute phase and stable phase based on clinical efficacy and thedynamic CSF cytology monitoring situation.25patients with no centralnervous system infections in the same period were selected for the controlgroup. All patients including the acute phase and stable phase of CNM andcontrol group take the lumbar puncture to collect the cerebrospinal fluid.75copies of CSF were analyzed CSF routine、biochemistry and measured thelevels of Th1cytokine IFN-γ, TNF-α and Th2cytokines IL-10proteins in theCSF by ELISA.Results:1Clinical data analysis:25cases of CNM patients wereconfirmed by cerebrospinal fluid pathogen detection, use CSF MGG jointAllie new staining. Among them,16were male and9were female, aged from 6to78years, an average of42.12±2.76years. The control group, a total of25cases,12males and13females, aged from6to58years with an average of16.32±13.18years. CNM compared with the control group, the total numbersof cells, white blood cells and protein content in CSF were increased, whileglucose and chloride content in CSF decreased. The differences werestatistically significant (P <0.05). CNM stable phase compared with acutephase, CSF pressure, the total number of cells, white blood cell count andprotein content were lower, while CSF glucose and chloride content werehigher (P <0.05).2Dynamic CSF cytology monitoring of CNM patients: Cryptococcus isin heaps in the CSF testing of acute phase and the neutrophil is the mainbackground inflammatory cell. The number of Cryptococcus decreasesdramatically in stable phase. We can see cryptococcal phagocytein the MGGstaining and lymphocyte is the main background inflammatory cell.Wemonitored the cerebrospinal fluid cytology dynamically and found that theCryptococcus number in CSF was significantly reduced in11~35days,average21days after treatment.3Cytokines in the CSF of CNM and control patients: The IFN-γ, TNF-αand IL-10concentrations were measured as11.14pg/ml±1.45，18.48pg/ml±3.07and44.04pg/ml±10.16,respectively, in acute phase CNMpatients. In20control patients, the levels of these cytokines were13.82pg/ml±1.4,28.52pg/ml±3.57and7.78pg/ml±1.32,respectively. IFN-γ,TNF-α and IL-10are significantly different between acute phase CNMpatients and controls (p<0.01, p<0.01and p<0.01,respectively). The level ofIFN-γ and TNF-α are significantly lower in CNM patients than in controlpatients, while the level of IL-10are significantly higher in CNM patients thanin control patients.4Levels of cytokines in CSF from acute phase and stable phase CNMpatients: The CSF IFN-γ, TNF-αand IL-10levels were quantified and foundto be11.14pg/ml±1.45，18.48pg/ml±3.07and44.04pg/ml±10.16, respectively,in acute phase CNM patients. The CSF levels of these cytokines were17.64pg/ml±2.29,49.40pg/ml±9.31and23.06pg/ml±7.49, respectively, inthe stable phase. The IFN-γ, TNF-αand IL-10levels in acute phase CNMpatients are significantly different from the levels seen in stable phase patients(p<0.01, p<0.01and p<0.01, respectively).IFN-γ and TNF-αare significantlylower in acute phase CNM patients compared to stable phase patients, butIL-10are significantly higher in the acute phase.5Distribution of Th1-Th2responses in CNM patients: The levels of theratio of Th1/Th2were used to assess the contribution of Th1-Th2immunity inacute and stable phase CNM patients. The ratios of IFN-γ/IL-10and TNF-α/IL-10measured in CSF were significantly lower in acute phase CNM patientsthan in the CSF of control patients (p<0.01and p<0.01, respectively). Theseratios were also significantly lower in acute phase patients compared to stablephase patients (p<0.01).Conclusion: In25Cryptococcus meningitis patients, IL-10was high,while the IFN-γ/IL-10and TNF-α/IL-10ratios were low. These results revealthat the Th2response is dominant in patients with C.neoformans infection.The IFN-γ/IL-10and TNF-α/IL-10ratios were significantly higher in stablephase compared to acute phase CNM patients, which shows that the immuneresponse shifted towards to a Th1response in the stable phase and providesdirect evidence that the imbalance of Th1-Th2cytokines is related to thepathogenesis of C.neoformans. It also proves that cellular immune responsesare the major defense against C.neoformans infection.