Role of circulating and tumor associated immune cells infiltration in triple negative breast cancer (tnbc) and their association with inflammatory and hypoxic markers

Aim: To study the role of circulating and tumor associated immune cells infiltration in Triple Negative Breast Cancer (TNBC) and their association with inflammatory and hypoxic markers.
Methods: In this study, 50 Triple Negative Breast Cancer (TNBC) patients and 25 healthy controls were enrolled. Of them, differential WBC count was available in only 34 TNBC patients. Neutrophil count, Lymphocyte count, Neutrophil to Lymphocyte ratio (NLR) and Platelet to Lymphocyte ratio (PLR) was studied in these 34 TNBC patients and compared with healthy controls. CD3+ T cells, MPO+ Neutrophils as well as hypoxic markers IL8, HIF-1and TNF-α were evaluated in all 50 TNBC patients by immunohistochemistry (IHC) method. 
Results: In comparison to healthy controls, mean percentage of neutrophils and NLR was significantly higher in TNBC patients, whereas mean percentage of lymphocytes (p=0.002) was significantly lower in TNBC patients. In relation to clinicopathological variables, increased neutrophils, high NLR and decreased lymphocytes were associated with large tumor size, lymph node positivity and advance stage. Further, high PLR was significantly associated with lymph node positivity. Regarding tumor associated immune cells, patients with high tumoral and stromal infiltration of CD3+ T cells was associated with histological grade III tumor and advanced stage. Similarly, MPO+ neutrophils infiltration in tumor and stroma tended to be high in patients with premenopausal status and large tumor size. Further, with respect to inflammatory cytokines and hypoxia related markers, IL-8 expression was noted high in patients with young age and larger tumor size. HIF-1 expression was significantly higher in grade III tumors and significant high incidence of tumoral TNF-α expression in tumor was noted in patients with T3 tumor size. 
Conclusion: The present study observed lymphopenia and neutrophilia with high NLR, high PLR and its association with disease aggressiveness. Further, infiltration of CD3+ T cells in tumor microenvironment was seen in tumors with advanced histological grade of the tumor, whereas tumor infiltrating neutrophils, TNFα and IL-8 may be cytotoxic to tumor cells via an oxygen radical dependent mechanism.