CD163 Tumor-Associated Macrophages Long-Term Prognostic Value In Locally Advanced Nasopharyngeal Carcinoma

Purpose:There is a complex tumor microenvironment in nasopharyngeal carcinoma.With the deepening research on TME,TAMs play an important role in the pathogenesis and development of tumor s.TAMs can be divided into M1 type and M2 type macrophages.CD68 is the associated antigen of M1 macrophages and M2 macrophages.CD163 is a specific antigen associated with M2-type macrophages.In this study,the expression of CD163 and CD68 immunohistochemical double staining in nasopharyngeal carcinoma was used to analyze the relationship between the degree of infiltration of different types of TAMs and clinical features,and to explore the correlation between the degree of infiltration of different types of TAMs and the prognosis of nasopharyngeal carcinoma.Method:A total of 239 patients with nasopharyngeal carcinoma admitted to our hospital from January 2014 to January 2017 were selected.All the histopathologic types were non-keratinized squamous cell carcinoma.Three groups of tumor -related macrophages were marked by CD163 and CD68 automatic immunohistochemical double staining in paraffin sections of pathological tissues.CD163+CD68+TAMs(M2),CD163+CD68-TAMs and CD163-CD68+TAMs;Panoramic scan of pathological sections;HALO digital pathological image analysis.The HALO digital pathological image analysis software platform was used to conduct a comprehensive quantitative analysis of CD163 expression,CD68 expression and three groups of tumor -related macrophages,and the positive area ratio was obtained.The optimal cut-off value was obtained by X-tile analysis software based on DMFS follow-up data.Kaplan-Meier method was used to calculate the survival rate,log-rank was used to analyze the influence of different factors on prognosis,rank sum test and Chi-square test were used to analyze the correlation between CD68,CD163 and tumor -related macrophages in the three groups and clinical characteristics,and Spearman was used to analyze the correlation between tumor macrophage infiltration degree in the three groups.Cox regression model was used to analyze the correlation between different factors and the prognosis of nasopharyngeal carcinoma.Statistical software SPSS(version 26.0)produced by IBM was adopted for data analysis.The study used R-language C-Index method to compare the degree of infiltration of CD163+CD68+TAMs and CD163+CD68-TAMs in the prognostic value of nasopharyngeal carcinoma(windows version).Results:1 The median follow-up of the whole group was 64 months,ranging from 2 to96 months.During follow-up,27 patients(11.3%)died,18 patients(7.5%)recurred,and 24 patients(10%)developed distant metastases.5-year overall survival rate was87.2%,5-year progression-free survival rate was 75.3%,5-year relapse-free survival rate was 90.5%,and 5-year distant metastasis-free survival rate was 87.8%.2 Kaplan-Meier analysis showed that patients with low infiltration of CD163+CD68+TAMs had 5-year OS(93.6% vs.82.8%,P=0.001),、5-year PFS(85%vs.60%,P<0.001),、5-year DMFS(95% vs.75.5%,P<0.001)were higher than those with high infiltration.Multiple factors proved that the infiltration degree of CD163+CD68+TAMs could be an independent risk factor for OS,PFS and DMFS(P=0.012,P=0.002,P=0.005).The 5-year OS(90.5% vs.86.3%,P=0.024)and 5-year DMFS(91% vs.79.3%,P=0.017)of patients with low infiltration of CD163+CD68-TAMs were higher than those with high infiltration.There was no correlation between the infiltration degree of CD163-CD68+TAMs and OS,PFS,RFS and DMFS(P>0.05).3 Chi-square test analysis showed that the proportion of CD163+CD68+TAMs and CD163+ CD68-TAMs with high infiltration degree was higher in males than in females(P<0.001),higher in patients aged >60 years than in patients aged ≤60 years(P<0.001),higher in patients with stage IV than in patients with stage III(P=0.008),and higher in plasma EBV-DNA Positive patients were higher than negative patients(P<0.001).The proportion of CD163-CD68+TAMs with high infiltration degree was higher in patients ≤60 years old than in patients >60 years old(P<0.001),and in patients with stage III than in patients with stage IV(P=0.042).There was no correlation between the proportion of CD163-CD68+TAMs with high infiltration degree and sex or plasma EBV-DNA(P>0.05).4 In univariate analysis of clinical features and prognosis of nasopharyngeal carcinoma,OS in patients IV was significantly worse than that in patients with stage III(HR=0.382,P=0.023).The RFS of patients with ≥2 chemotherapy cycles were higher than those with <2 chemotherapy cycles(HR=5.410,P=0.026).Multiple factors confirmed that clinical stage was an independent factor for patients’ OS(P=0.044),while chemotherapy cycle was not an independent factor for patients’ RFS(P=0.080).Age,sex,plasma EBDNA and LDH were not prognostic factors.5 Using C-Index,the study showed that the infiltration degree of CD163+CD68+TAMs was superior to that of CD163+ CD68-TAMs in the evaluation of prognosis.Conclusion:The degree of CD163+CD68+TAMs(M2)infiltration could be an independent risk factor for OS,PFS and DMFS(P<0.05).It is suggested that the infiltration degree of CD163+CD68+TAMs is a predictor of overall survival,progression-free survival and distant metastasis risk of nasopharyngeal carcinoma.