| Objective:to evaluate different tumor-infiltrating immune cells in breast cancer patients after neoadjuvant chemotherapy through H&E staining and immunohistochemical assays for CD68and CD8, and determine whether these cells have correlation with patients’outcome.Methods:We carried out H&E staining and immunohistochemical assays for CD8and CD68markers on formalin-fixed paraffin-embedded invasive breast cancer samples from patients diagnosed between Jan.1st2003and Dec.31st2006in Shanghai Cancer Center. And all of these samples were residual tumors after neoadjuvant chemotherapy.Results:120patients considered as non-pCR responders were included in this study. The positivity rate of TIIC in non-luminal patients was higher than luminal ones (P=0.034). The positivity of adjacent stromal CD8+T cells was inversely correlated with the expression of Ki-67(P=0.001).The positivity of adjacent stromal and distant stromal CD8+T cells were positively correlated with lymphovascular invasion (LVI)(P values were0.045and0.013, respectively). The positivity of intra-tumoral macrophages was positively correlated with tumor size pre-and post-NCT and nodal status after NCT (P values were0.004,0.049and0.020, respectively). Moreover, the positivity rate of adjacent stromal macrophages was positively correlated with Ki-67index, LVI and nodal status after NCT (P values were0.016,0.011and0.027, respectively). And the positivity rates of adjacent stromal macrophages were different between each other in all kinds of breast cancer molecular subtypes (P=0.049). What’s more, the positivity of distant stromal macrophages was significantly associated with Ki-67index, the primary tumor size and HER-2expression (P values were0.024,0.045and0.043). Combining clinical and pathological features of all patients, only the expression of adjacent stromal macrophages was correlated with clinical response of NCT (P=0.009). In univariate analysis, TIIC-positive patients had a significantly better prognosis than the negative ones (P=0.006). The total CD8+T cell-positive or adjacent stromal CD8+T cell-positive patients had a longer DFS than the respective negative ones (P values were0.006and0.026). In the subgroup analysis, adjacent stromal CD8+T cell-positive patients had a better outcome than the negative ones in HER-2+, ER/PR-,N2,N3,pN3,T3,T4, or LVI-positive patients (P values were0.001,0.008,0.043,0.004,0.014,0.008,0.021,0.018,0.001). Although there wasn’t a significant correlation between DFS and distant stromal CD8+T cells, we found that the distant stromal CD8+T cell-positive patients had a longer DFS in ER/PR-, HER-2+or non-luminal patients (0.037,0.007,0.030and0.033). As to macrophages, we found that the positivity of macrophages in these three locations indicated a worse prognosis. However, only the adjacent stromal ones reached a statistical significance (P=0.032). Considering all the other clinical and pathological parameters, pN grading (P=0.002), Ki-67expression after NCT (P=0.001),TIIC (P=0.006), total CD8+T cells (P=0.006), the adjacent stromal CD8+T cells (P=0.026), the adjacent stromal macrophages (P=0.032) had correlation with DFS in univariate analysis. However, in multivariate cox regression analysis, pN grading (P=0.007, HR:1.583,95%CI1.136-2.206), Ki67(P=0.010, HR:2.335,95%CI1.222-4.463), total CD8+Tcells (P=0.032, HR:0.332,95%CI0.122-0.908), the adjacent stromal macrophages (P=0.027, HR:1.968,95%CI1.081-3.583) and TIIC (P=0.011, HR:0.421,95%CI0.216-0.823) were proved to be independent predictors of DFS.Conclusions:The infiltrating immune cells in breast cancer after NCT were correlated with the clinical and pathological features of patients. More importantly, the total tumor-infiltrating immune cells、 total CD8+T cells and the adjacent stromal macrophages were proved to be independent predictors of patients outcome, which offered a deeper insight of tumor microenvironment and could allow physicians to provide treatment strategies specifically and individually. |
PDF Full Text Request