The Research Of Changes Of Some Immune Cells And Immune Checkpoints In Tumor Immune Microenvironment Before And After Treatment With PD-1 Inhibitors

[Objective]To collect and investigate the changes in the tumor immune microenvironment of the tumor tissue samples between before and after the treatment of the PD-1 inhibitor,and to analyze the influence of the pre and post treatment indicators and the changes of the index on the survival and efficacy of the patients before and after the treatment,in order to explore the treatment of PD-1 inhibitors.A new theoretical basis is sought to explore the mechanism of PD-1 inhibitors for the treatment of acquired resistance,and to provide clues for the selection of treatment options for the treatment of PD-1 inhibitors.[Method]The patients with advanced malignant solid tumors treated with PD-1 inhibitors were collected to summarize the patient’s clinical data and follow up their survival information.Multiple labeling immunofluorescence staining,spectral imaging and signal resolution techniques,cell,tissue phenotypic.identification and quantitative statistics were used to compare the paired tumor before and after the treatment of PD-1 inhibitors.The changes in the tumor immune microenvironment of the tissue samples,including the immune cell markers CD8,CD33,FoxP3 and immunological checkpoints OX40,ICOS,Tim3,LAG3,were used to analyze the effects of pre and post treatment indicators and the changes of indexes before and after treatment on the survival and efficacy of the patients.[Result]1.A total of 25 patients were enrolled and the tumor tissue samples were collected both before and after the treatment of PD-1 inhibitors.The median progression free survival time for 25 patients was 4.90 months,and the 95%confidence interval was 3.11 to 6.695 months.Median overall survival time has not yet been achieved.The objective response rate was 8%,and the disease control rate was 72%.2.After the treatment of PD-1 inhibitors,the expression of CD8 was significantly decreased(P=0.0161),the expression of CD33 was significantly increased(P=0.0412),and there was no statistical difference in FoxP3(P=0.0687).All the 25 patients were divided into two groups according to the therapeutic effect,respectively,the progression group and response group,and CD8,CD33 and FoxP3 were no longer than before the treatment.Obvious difference(progression group:P=0.0725,P=0.1185,P=0.1369;response group:P=0.0546,P=0.1502,P=0.3307).3.Before the treatment of PD-1 inhibitors,there was no significant difference in CD8,CD33,and FoxP3 between the progression group and response group(P=0.6887,P=0.9133,P=0.1674),and the CD8,CD33 and FoxP3 between the two groups were still not statistically different after the treatment of PD-1 inhibitors(P=0.1958,0.1014).There was no significant difference in the value(P=0.7567,P=0.1105),but there was a difference in the CD33 value.The CD33 in the progression group had a significant change(P=0.0297).4.Before the treatment of PD-1 inhibitors,the level of CD8 and CD33 did not affect PFS(P=0.6274,P=0.7750).The PFS of the patients with low expression of FoxP3 before treatment was significantly longer than that of the high expression patients(P=0.0310),and the CD8 level had nothing to do with PFS..0034);the changes of CD8,CD33 and FoxP3 did not affect PFS(P=0.4722,P=0.3883,P=0.3216)before and after treatment.5.There was no significant difference in the changes of OX40 and ICOS after the treatment of PD-1 inhibitors(P=0.99 68,P=0.5998).All 25 patients were divided into two groups according to the therapeutic effect,respectively,the progression group and response group.There was no significant difference between OX40 and ICOS before the treatment(progression group:P=0.3629,P=0.5741,and response group:P=0.7793,P=0.6327).6.Before the treatment of PD-1 inhibitors,there was no significant difference in OX40 and ICOS between the progression group and response group(P=0.6465,P=0.2189),and there was no statistical difference between the two groups after the treatment of PD-1 inhibitors(P=0.7294),but the ICOS in response group was significantly higher than progression group(P=0.0485),and the OX40 and ICOS in the two group had no significant difference in OS value(P=0.4231,P=0.7149).7.The levels of OX40 and ICOS did not affect PFS(P=0.1813,P=0.0691,P=0.6848,P=0.4056)before and after the treatment of 7.PD-1 inhibitors,and the values of OX40 and ICOS before and after treatment did not affect PFS(P=0.9075,).8.The expression of Tim3 and LAG3 increased significantly after the treatment of 8.PD-1:inhibitors(P=0.0172,P=0.0296),and there was no statistical difference in PDL1(P=0.2988).All 25 patients were divided into two groups according to the therapeutic effect,which were the progression group and response group.PDL1 before treatment of the two groups had significant difference(P=0.219).There was no significant difference between Tim3 and LAG3 before the treatment(P=0.1961,P=0898),and there was no significant difference between PDL1 and LAG3(P=0.5282,P=1873)in the response group(P=0.5282,P=1873)after progression(P=0.5282,P=1873),and significantly higher than before the treatment(P=0.0260).9.Before the treatment of 9.PD-1 inhibitors,the PDL1 of the progression group and response group were significantly different,the latter was significantly higher than the former(P=0.0155).There was no statistical difference between the two groups(P=0.0653,P=0.3757),and there was no statistical difference between the PDL1,Tim3 and LAG3 between the two groups after the treatment of PD-1 inhibitors(P=0.0598,0679,P=0.2009);there was no significant difference in the PDL1,Tim3 and LAG3 values between the two groups(P=0.7914,P=0.2456,P=0.1707).10.Before the treatment of 10.PD-1 inhibitors,the level of Tim3 and LAG3 did not affect PFS(P=0.13713 P=0.2558).The PFS of the patients with low expression of PDL1 before treatment was significantly shorter than that of the high expression patients(P=0.0035).After the PD-1 inhibitor treatment,PDL1 and Tim3 levels were significantly longer than those of the high expression.0123);before and after treatment,the PFS of PDL1 decreased significantly,and the change of Tim3 and LAG3 did not affect PFS(P=0.0755,P=0.4782).[Conclusion]1.The decrease in the number of cytotoxic CD8+T cells and the up-regulation of inhibitory components in the microenvironment,including inhibitory cell CD33+MDSCs and inhibitory immunologic checkpoint Tim3,LAG3,are the manifestations of the deterioration of the tumor immune microenvironment.The progression of the PD-1 inhibitor treatment failure is closely related to the deterioration of the tumor immune microenvironment in the patients,and this also suggests possible treatment options after drug resistance of PD-1 inhibition.2.During the treatment of PD-1 inhibitors,the expression of the stimulating immunologic checkpoint(0X40,ICOS)and the inhibitory immunologic checkpoint(Tim3,LAG3)was in a dynamic change,and the effect of the irritant immunologic checkpoint could be improved.On the contrary,the up-regulation of the inhibitory immunologic checkpoint indicated that the therapeutic effect was poor.3.The level of PDL1 before treatment can affect the efficacy of PD-1 inhibitor treatment,which is consistent with previous studies.4.Before treatment,FoxP3 expression density and PDL1 expression level can influence the prognosis of patients,especially FoxP3 can be used as a potential index for screening PD-1 dominant population.