The X-ray structure of the PD-1/PD-L1 complex was downloaded from your protein data bank website. and found that PAC-1 a high immune score and M2 TAMs were strongly associated with poor clinical outcomes in patients with MIBC. Further, we analyzed resected samples from 120 patients with MIBC and found that individuals with PD-1-positive TAMs showed a reduction in 5-12 months overall survival and disease-free survival. Additionally, PD-1-positive TAMs showed a significant association with higher programmed death-ligand 1 (PD-L1) expression, the Ki67 index, the pT stage and fewer CD8-positive T cells. Through the co-immunoprecipitation (co-IP) assay of THP-1 derived macrophages, we found that CD68 can bind to PD-1. The binding of CD68 and PD-1 PAC-1 can induce M2 polarization of THP-1 derived macrophages and promote malignancy growth. The anti-CD68 treatment combined with peripheral blood mononuclear cells (PBMC) showed obvious synergy effects on inhibiting the proliferation of T24 cells. Together, these results indicate for the first time that CD68/PD-1 may be a novel target for the prognosis of patients with MIBC. KaplanCMeier analysis and the log-rank test. The prognostic significance of the clinicopathological parameters was analyzed using the chi-square test. The Spearman correlation analysis was used to analyze the correlation between CD68, PD-1, and PD-L1. The relationship of PD-1-positive TAMs CD8-positive T cells was analyzed by Students a LASSO analysis, and recognized 8 survival-associated immune cell types. Further, we derived an immune score for predicting the prognosis of patients with MIBC ( Figures 1A, B ). According to the Kaplan-Meier analysis of the TCGA cohort, PAC-1 patients with MIBC having high immune scores were associated with a significantly reduced 5-12 months OS and DFS outcomes ( 0.0001 and 0.0001, respectively; Figures 1C, D ). The heatmap of the survival, tumor stage, tumor grade, immune score, and the profiles of different immune cells is shown in Physique 1E . Interestingly, M2 TAMs showed a correlation with high immune scores in the heatmap ( Physique 1E ). Therefore, we further analyzed the clinical outcomes for patients with PAC-1 MIBC showing the presence of M2 TAMs in the TCGA cohort. Patients with M2 TAMs showed significantly worse 5-12 months OS and DFS outcomes ( 0.001, respectively; Figures 3B, C ). The 5-12 months OS rate was estimated to be ~33.3% Rabbit polyclonal to TDGF1 in patients with PD-1-positive TAMs, and ~77.78% in patients with PD-1-negative TAMs ( Figure 3B ). Additionally, the 5-12 months DFS rate was ~21.1% in patients with PD-1-positive TAM compared with ~66.7% in patients with PD-1-negative TAM ( Determine 3C ). Open in a separate window Physique 3 (A) Immunohistochemical staining of PD-1-positive TAMs (reddish arrow), PD-1-unfavorable TAMs (purple arrow), strong PD-L1 and poor PD-L1 expression. (B)?Kaplan-Meier analysis of PAC-1 OS in patients with MIBC with PD-1-positive TAMs in the Shanghai General Hospital cohort. (C) Kaplan-Meier analysis of DFS in patients with MIBC with PD-1-positive TAMs in the Shanghai General Hospital cohort. (D) PD-1-positive TAMs showed less CD8-postive T cells nearby. *** 0.001 (Students = 0.027, Table 1 ). Patients with PD-1-positive TAMs were found to exhibit higher pT stages than those without. PD-1-positive TAMs also showed significantly stronger PD-L1 expression ( 0.001), a higher Ki67 index (= 0.003), and worse pathological patterns ( 0.001; Table 1 ). Interestingly, higher PD-L1 expression levels also resulted in poor prognosis of patients with MIBC (data not shown). We investigated the response to cisplatin-based neoadjuvant chemotherapy in patients with PD-1-positive and -unfavorable TAM phenotypes. Patients with MIBC who were administered neoadjuvant chemotherapy in the pT2 stage showed a better prognosis. However, the presence of PD-1-positive and -unfavorable TAM did not improve the response to neoadjuvant chemotherapy ( 0.05, data not shown). Patients with a PD-1-positive TAM phenotype showed a comparatively substandard response to neoadjuvant chemotherapy, which was similar to the 5-12 months OS and DFS outcomes in patients with PD-1-positive TAMs. Intriguingly, PD-1-positive TAMs showed relevance to bladder malignancy related immune response. Based on 120 MIBC patients from Shanghai General Hospital, we found that CD8-positve T cells were comparatively fewer around PD-1-positive TAMs ( 0.001; Physique 3D ), indicating PD-1-positive TAMs could be involved in bladder cancer immune response. In addition, the number of PD-1-positive TAMs showed positive relevance to the PD-L1 expression of bladder malignancy cells (= 0.48, 0.001; Physique 3E ). The Conversation of CD68 and PD-1 Induced TAMs to M2 Polarization Interestingly, when we further analyzed the presence of PD-1-positive TAMs using immunofluorescence staining on FFPE samples, CD68 and PD-1 tended to be expressed synchronously in TAMs ( Physique 4A ). Hence, we conducted an analysis for the correlation of CD68 mRNA expression levels with the PD-1 and PD-L1 mRNA expression levels in the TCGA cohort and found that the expression of CD68 and that of PD-1 and PD-L1 was correlated (= 0.58 and = 0.41, respectively; 0.001 and .
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