Flow Cytometry To Identify ??T Cells In Breast Cancer And Its Clnicopathological Significance

Objective:1. Find ??T cells in breast tumor-infiltrating lymphocytes and peripheral blood PBMC By flow cytometry method;2. analyze the relationship between ??T cells and clinical pathologic factors.3. Preliminary Explore the secretion of IL-17 in human breast cancer.Methods:We collected 41 breast cancer patients with fresh human breast cancer tissue specimens, and their normal breast tissue specimens and blood samples. 30 normal human peripheral blood samples were collected too. Extracted the lymphocytes in each group. ??T cells were labeled by CD3- APC and ??TCR-Per CP-Cy TM5.5 two antibodies. Using flow cytometry method to measure the contents of ??T cells in each group, statistical comparisons and analyze their relationship with clinicopathological factors of breast cancer. Take 10 of 41 cases of breast cancer cells suspension,Using CD3-APC and IL-17-PE two flow-type antibody labeled CD3~+ cells and cells that secrete IL-17.Results:1. CD3~+ cells were found in all 41 cases of breast cancer tissues, the average ratio was 33.25%, while only 9 cases of normal breast tissue found CD3~+ cells, with an average ratio of 1.41%, the difference was significant(P <0.05); The CD3~+ T cells in breast cancer tumor infiltrating lymph cells were correlated with the breast cancer molecular typing,tumor grade, lymph node involvement;2. ??T cells were found in all 41 cases of breast cancer tissues, the average ratio was 10.72%, while only 3 cases of normal breast tissue samples detected ??T cells, with an average ratio of 0.23%, the difference was statistically significant(P <0.05). Breast tumor infiltrating lymphocyte ratio of ??T cells was related with molecular typing pathological stage, Lymph node metastasis, HER-2 expression significantly(P <0.05), but nothing to do with the patient's age, tumor size, ER and PR expression(P> 0.05).;2. we found ??T cells in peripheral blood PBMC of patients with breast cancer with the average ratio of 6.22%. The ratio in normal human peripheral blood PBMC was 4.44%. The difference was significant(P <0.05). But The proportion of ??T cells in peripheral blood of patients with breast cancer are irrelevant with age, tumor size, pathologic stage, lymph node metastasis, ER, PR,HER-2 expression and the Content of ??T cells in tumor. Breast cancer CD3~+ cells content was positively correlated with ??T cells;4. 4 cases of 10 cases of tumor cell suspension secret IL-17, indicating that the breast cancer tumor infiltrating CD3~+ lymphocytes secrete IL-17, rather than the tumor cells.Conclusion:1. CD3~+ T cells in breast cancer tumor infiltrating lymphocytes was significantly higher than that in normal breast tissue and correlated with breast cancer molecular typing,tumor grade and lymph node involvement;2. The ??T cells in breast cancer tumor infiltrating lymphocytes was significantly higher than that in normal breast tissue and positively correlated with breast cancer molecular typing,tumor grade, lymph node involvement and HER-2 expression;3. The ??T cells in peripheral blood of breast cancer patients is higher than that in normal person, but no significant correlation with the clinicopathological parameters of breast cancer and the Content of ??T cells in tumor. CD3~+ cells content of breast cancer was positively correlated with ??T cells;4. tumor infiltrating lymphocytes rather than the tumor cells secrete IL-17.