Identification Of Changes Of Tumor Immune Microenvironment And Its Clinical Implication Before And After Neoadjuvant Chemotherapy In Cervical Cancer

Objective: To investigate the changes of tumor immune microenviroment in cervical cancer patients treated with neoadjuvant chemotherapy(NACT),and to analyze the correlations between immune features,NACT response and clinical prognosis of patients.Methods: Cervical cancer patients with FIGO stage IB3,IIA2,IIB or above were collected,and all patients were treated with NACT before radical surgery.Clinical information of patients was collected,and prognostic information was obtained through follow-up visits.The relationships between clinical factors and NACT response and clinical prognosis of patients were analyzed.The paraffin sections of tumor tissues before and after NACT were obtained,and the expression of CD3,CD8,CD4,Fox P3,CD68,CD11 c,IDO-1,PD-1,PD-L1,cytokeratin(CK)and P16 before and after NACT were investigated by multiple immunohistochemical techniques.The changes of each indicator before and after NACT treatment,and the relationship between each indicator and patients’ NACT response and clinical prognosis were explored.Results: 1.A total of 132 patients were included in this study,all of whom received 1 to 2cycles of NACT before radical surgery.During follow-up,24 patients had recurrence/metastasis,and 18 patients died of cancer.The median progression-free survival of patients was 23.5 months,and the median overall survival was 25 months.2.Among the 132 patients received NACT,85(64.4%)had a response and 47(35.6%)had no response.Logistic regression found no clinical factors associated with NACT response.Cox regression was used to analyze the correlation between clinical characteristics,surgical pathology and patient prognosis.The results indicated that intrauterine metastasis was the only factor associated with PFS(HR 6.853,95%CI 1.418-33.116,P=0.017)and OS(HR11.077,95%CI 2.147-57.156,P=0.004).3.Seven-color immunohistochemical techniques were used to explore the changes of immune features before and after NACT(A total of 92 patients had matched specimens before and after NACT).A total of 11 indicators were divided into 2 panels.Panel 1: CD8,CD4,Fox P3,PD-1,PD-L1,CK,and DAPI.Panel 2: CD3,CD68,CD11 c,IDO-1,P16,CK,and DAPI.The results suggested that NACT can promote the infiltration of CD3+,CD4+and CD8+ T cells in stromal,tumor and overall region,reduce the proportion of Treg and CD4+ T cells,and reduce the proportion of CD11c+IDO+ positive cells in CD11c+ cells.In tumor areas,NACT decreased the positive rate of P16.4.The differences of indicators among patients with different NACT responses were analyzed.The results indicated that in stromal,tumor and overall region,CD3+ T cells were highly expressed and CD8+ T cells were lowly expressed in respond patients before NACT.Lasso regression and Logistic regression were used for the Nomogram,and the results showed that highly expressed CD3+ in the stromal and tumor,and lowly expressed CD8+ in stromal and tumor,indicated that the patient had a good response to NACT treatment.After NACT,CD8+ T cells in patients with response increased significantly compared with that before treatment,and the decrease of Treg in tumor and overall areas was more obvious.The Nomogram by Lasso regression and Logistic regression showed that significantly increased CD8+ after treatment,and decreased Treg in the tumor after treatment,indicated response of NACT treatment.5.The differences of indicators among patients with different prognosis were analyzed.The results indicated that before NACT treatment,CD4+ and CD8+ T cells,Treg,and PD-1 in patients with recurrence were significantly lower than those without recurrence.After NACT treatment,the proportion of Treg in CD4+ T cells did not significantly decrease in relapsed patients compared with pre-NACT treatment.Lasso regression and Logistic regression were used for the Nomogram,and the results showed that the positive rates of CD8+,Treg,Treg/CD4+ and CD8+PD-1+/ CD8+ in the stroma region before treatment were of high prognostic value for relapse.Conclusions: 1.NACT treatment of cervical cancer can cause changes in the microenvironment of cervical cancer,mainly including promoting the infiltration of CD3+,CD4+ and CD8+ T cells,reducing the proportion of Treg and Treg in CD4+ T cells,and reducing the proportion of CD11c+IDO+ positive cells in CD11c+ cells.2.The high expression of CD3+ in the stroma and tumor regions,the low expression of CD8+ in the stroma and tumor regions,the significant increase of CD8+ after treatment,and the significant decrease of Treg in the tumor region after treatment all suggest that the patient has a good NACT response.3.The low expression of CD8+,Treg,Treg/CD4+ and CD8+PD-1+/CD8+ in the tumor stroma region of patients before NACT treatment suggests a poor prognosis.