Expression Of Microenvironment Related Factors In DLBCL And Its Clinical Significance

Background:Diffuse large B-cell lymphoma(DLBCL)is the most common type of non-Hodgkin's lymphoma.Its incidence in the adult population accounts for 30%of the total lymphoma and more than 40%in Asia.International prognostic index(IPI score)is the main DLBCL clinical prognostic indicators.As lymphoma therapy entered the era of rituximab,the IPI scoring system was adjusted with a variety of indicators.Molecular typing based on immunohistochemistry and gene detection was evaluated for its value to the prognostic system.In addition,the study of tumor microenvironment related factors has gradually received attention.The clinical significance of some of these factors'characteristic expression in lymphoma microenvironment and its relationship with clinical prognosis deserve further exploration.Intent:In this study,we examined the expression of CD163,VEGF,PD-1/PD-L1 in DLBCL tissue.Clinical information and survival follow-ups were collected to clarify the relationship between DLBCL clinical pathological features and prognosis,and the clinical significance was analyzed.Methods:Pathologically confirmed cases of patients with diffuse large B-cell lymphoma treated in the hospital from January 2009 to June 2017 were collected.Incomplete clinical cases and cases that were unable to obtain pathological tissue were removed.Case collection process was approved by the Ethics Committee.After getting enough tissue sections,immunohistochemical method were used to detect the expression of CD163,VEGF,PD-1/PD-L1 in tumor cells and tumor microenvironment separately.And we statistically analyzed its correlation with relevant clinical indicators using?~2 test and other statistic method.Result:1.A total of 49 cases of patients with newly diagnosed diffuse large B celllymphoma were collected in our hospital,including 28 males(57.1%)and 21 females(42.9%),?60 years old 31 of them(63.3%)were over 60 years old and 18 of them(36.7%)were under 60.The median age was 62 years old(range from 32 to 83).As for their Ann Arbor stage,I-II in 25 cases(51.0%),III-IV of 24 cases(49.0%).Lactate dehydrogenase(LDH)normal in 23 cases(46.9%),elevated in 26 cases(53.1%).26cases(53.1%)had extranodal invasion,23 cases(46.9%)had no extranodal invasion.There were 12 patients(24.5%)with 0-1 points in IPI score,30(61.2%)with 2-3 points and 7(14.3%)with 4-5 points.There were 14 cases(28.6%)of germinal center and 35cases(71.4%)of non-germinal center.ESR was normal in 13 cases(26.5%),elevated in36 cases(73.5%).28 cases(57.1%)had normal?2-microglobulin,and 21 cases(42.9%)were elevated.There were 16 patients(32.7%)with B symptoms and 33 patients(67.3%)without B symptoms.2.In the aspect of tumor microenvironment,35 cases(71.4%)were CD163positive for and 14(28.6%)were negative;32 cases(65.3%)were positive for VEGF and 17(34.7%)were negative;PD-1 positive in 9 cases(18.4%),negative in 40 cases(81.6%);PD-L1 was positive in 46 cases(93.9%)and was negative in 3 cases(6.1%).3.Correlation analysis of protein expression:The expression of PD-1 in tumor cells and tumor microenvironment correlated with the expression of PD-L1?CD163?VEGF(P=0.000),which shows statistical significance.The expression of PD-L1 in tumor microenvironment also correlated with the expression of CD163?VEGF(with the P value of 0.003 and 0.000 respectively).There was no correlation between the expression of CD163 and VEGF(P>0.05).4.The expression of tumor microenvironment factors CD163?VEGF and PD-1/PD-L1 in the 49 newly diagnosed DLBCL patients was correlated with IPI score in pathological tissue(with the P value of 0.001,0.036,0.001,0.001 respectively),giving us statistical significance.However,there was no correlation between the expression of these factors and other clinicopathological features including gender,age,Ann Arbor stage,lactate dehydrogenase,extranodal invasion,whether germinal center,ESR,?2-microglobulin,and B symptoms.5.Due to insufficient follow-up time of patients diagnosed during 2017,5 cases were excluded.Follow-up of 44 patients diagnosed between 2009 and 2016 to December 31,2017,.Median follow-up:December 31,2017.Twenty patients(45.5%)died in 44 patients,24 cases(54.5%)survived.The median progression-free survival time was 11.92 months in the whole group,with a median overall survival of 19.45months.1-year progression-free survival and 1-year overall survival were calculated.6.Forty-four patients whose 1-year PFS and 1-year OS could be calculated were further analyzed for the relationship between microenvironment-related protein expression and survival.The results showed that there was a significant correlation between PFS and IPI score(P=0.015)and LDH(P=0.034).One-year OS was correlated with Ann Arbor stage(P=0.020)and VEGF expression in tumor cells(P=0.017),with statistical significance.In addition,the 1-year PFS and 1-year OS did not correlate with other patient parameters or the expression of CD163 and PD-1/PD-L1(P>0.05).Conclusions:1.High expression of CD163,VEGF and PD-L1 in tumor microenvironment of patientswithDLBCL,lowexpressionofPD-1andnegativecorre lation with the above microenvironment factor expression,suggesting a close relationship between macrophage and immunosuppression and angiogenesis in lymphoma microenvironment.2.DLBCL microenvironment-related factors CD163,VEGF,PD-L1,and PD-1were all associated with IPI prognostic score,suggesting that their expression may be related to prognosis,and worthy of further follow-up.