Effects And Mechanisms Of Anti-tuberculousis Treatment On Macrophages In Tuberculous Pleurisy

Objective:Tuberculosis pleurily occupies a large proportion in theextrapulmonary tuberculosis,the immune microenvironment in its typicalby macrophages can be observed from inducing delayed-typehepersensitivity.Macrophages in the body’s immune response tomycobacterium tuberculosis infection plays an important role,it is thefirst line of the body resistance to mycobacterium tuberculosisinfection,at the same time parasitism in the host. State of themacrophages are closely associated with the occurrence,development andoutcome of tuberculosis.Currently,standard therapy to treat tuberculosisstill used in clinical,the antibacterial effect and mechanism research ofdrugs has been very deep and clear.However,especially for durgs on theimmune cells of macrophage function if there is a regulation,the presentstudy is not deep.Only limited research stay in animal experiments,invitro experiment,the macrophages of peripheral blood as observationobject on the indirect evidence of a lack of againt human tuberculosiskichen system in this paper,the microenvironment.In view of this,thisstudy proposed based on tuberculos pleurity patients after treatment withtuberculosis pleural macrophage phenotypes,fuction and genedetection,as well as the in vitro drug direct stimulus to the pleuralmacrophages,to observe the above changes of pleural macrophages,discusses four kinds of anti-tb drugs have direct effect on macrophages and regulation mechanism,thus illustrating the role of macrophages in TBtreatment and mechanism,such as to clatify.Its targets,for the treatment oftuberculous pleurisy and outcome provide important theoretical basis foranalysis of the adjustment and prognosis of chemotherapy fortuberculosis and so on research and development of anti-tb drugs hasimportant clinical significance and value.Method:FMC detect macrophages percentage changes and the expression ofsurface markers;FMC detecting the percentage of macrophages beingswallowed by bead;ELISA method to detect tuberculosis pleural effusionafter treatment in the change of cytokines;On the depth sequencing ofmacrophages that before and after treatment;QPCR method is used todetect gene1s6110absolute quantitative analysis was carried out on thepleural effusion of tuberculosis bacterium;Anti-tb drugs single-agent andjoint processing in ivtro purified macrophages and detect the geneexpression level of macrophages.Result:In vivo：1After treatment with anti-tb drugs,patients with pleural surfacecarkers CD14, CD80and CD163and CD206fluorescence intensityincreased significantly,after three factors positive cell percentage alsoincreased.(MFI:p=0.017,p=0.0001, p=0.0001;Percentage:p=0.002,p＜0.0001,p=0.0003),CD86fluorescence intensity alsoelevated(MFI:P=0.004).2After anti-tb treatment, The percentage of T cell subsetCD3+CD4+and CD3+CD8+both reduced. Individual sample only had an incease or lower in CD3-CD19+and CD3-CD56+,but neither of themhad statisticallydifferences(P=0.578,P=0.424,P=0.602,P=0.923),CD3+CD4+/CD3+CD8+had no statistically differences.3The same sample by anti-tb drugs before/after treatment,theaverage percentage of multiple beads of phagocyte cells from79.74±8.30%to84.37±6.28%.4The level of proinflammatory factory of IFN-gamma andTNF-alpha in the pleural effusion was to reduce after theatment.(p﹤0.0001,p=0.045);Anti-inflammatory factorys IL-10were also reduced.(p﹤0.0001).5There are230DEG of macrophages in two groups before/aftertreatment,in which156gene expression level up adjustment and74genedown adjustment.Gene associated with bactericidal action COLEC12,CH13L1,FNBP1L expression are raised (p=0.0363,0.0022,0.0363);Macrophages of chemokine CCL18and CXCL5expressionincreased(p=0.0024,0.0013);Factor caspase5proinflammatory apoptosisexpression to decrease(p=0.0036);By interferon responses IFIT3,IFIT2and IFIT1experession were lower(p=0.0006,0.0015,0.0006);ChemokinesCXCL10and CCL8expression(p=0.0105,0.0155).In vitro：6The fluorescence intensity of each marker in the groups that dealedwith drugs has diffirent degrees elevate,CD14,CD80,CD86,CD206average fluorescence intensity of drugs in combination groups in the mostobvious(MFI:P=0.031,P=0.027,P=0.042,P=0.038).7Either rifampicin or pyrazinamide individual deal with cells,canenhance the expression of CD80and CD86(MFI:p=0.041,p=0.024forR,p=0.039,p=0.044for Z). 8Pyrazinamide or four drugs in combination with the Cmax doserespectively under the pleural macrophages CH13L1,mRNA expressionupward(p=0.038,0.042);Isoniazid and pyrazinamide can increase theexpression of FNBP1L,four drugs in combination can also make itexpress elevated(p=0.018,0.025,0.022),Pyrazinamide and four drugs incombination can increase the espression of CCL18(P=0.036,0.045).Thereis no difference in the result of PZA and four drugs in combination actingon the macrophage.Conclusion:1The standard anti-tuberculosis drugs in the treatment of tuberculouspleurisy,promote local pleural macrophage activation,lymphoid cells donot produce obvious effect on it.2Anti-TB drugs combined application,can be directly stimulate thepleural surface marker expression of macrophages, rifampicin andpyrazinamide played an important role.3Joint application of anti-TB drugs can directly enhance the pleuramacrophage phagocytosis,INH and pyrazinamide can promote geneexpression of CH13L1or FNBP1L isoniazid.4Pyrazinamide can promote the pleura macrophage chemotacticfactor CCL18experssion,can be associated with tuberculous pleurisypleural damage after the repair.5Joint application of anti-TB drugs,reduce the proinflammatoryfactor in the pleural effusion microenvironment IFN-gamma,TNF alphaand anti-inflammatory factor level of IL-10,and then cut the expression ofIFIT,can further inhibit macrophage for the release of proinflammatoryfactor,lead to the steady state lesion microenvironment.6The anti-tuberculous treatment in vivito changed significantly ofthe gene expression levels of macrophages of tuberculous pleurisy and the chang is closely related to the participate in the immune defensereaction,its mainly related gene expression up of the sterilization functionand down of INF mediated immune respose.