The Expression Of CD4~-CD8~-T Cell In Peripheral Blood Of Patients With Pancreatic Carcinoma And Its Biological Characteristics

Pancreatic carcinoma(PC) is one of the malignant tumor,its clinical manifestations was not obvious, developed rapidly, and its prognosis is very bad. At present,there is no effective screen and early diagnostic methods,it's often diagnosis at advanced stage.In developed countries,there are more than 20,000 person died of pancreatic carcinoma per year.In American,the median survival of patients with pancreatic cancinoma is only 6-18 months.In our country,the incidence rate of pancreatic carcinoma was similar with foreign countries.In 1990s the 5 years survival rates of shang hai crowed of men and women who with pancreatic carcinoma was 5.8% and 4.3% respectively.In recent years, the incidence and mortality rate of pancreatic cancer showed a rising trend year by year. Because of the specificity and location of pancreatic cancer and its early transfer infiltration,the opportunity of pancreatic carcinoma resection was reduced, and radiotherapy and chemotherapy the lack of positive effects, Therefore, the key to improving the survival rate of pancreatic carcinoma is to improve early diagnosis of pancreatic carcinoma and improving treatment of pancreatic carcinoma.In normal person,the percent of CD4-CD8- T cell (double negative T cell, DNT) in peripheral blood lymphocytes is 1%-5%,DNT cells may possibly derived from the CD8~+T and CD4~+ T cells,it can expression Fas/FasL,TNF/TNFR,CD3~+,CD4~-,CD8~-,MHCⅠ. DNT cells have important roles in immune regulation, immun suppression, autoimmune and anti-tumor activity. Now research shows that DNT cells have an important roles in anti-tumor activity,its may through Fas/FasL pathway to make tumor cell apoptosis. Another study showed that the cytokin like IFN-γsecreted by DNT cells may have an important roles in tumor therapy. We use flow cytometry analyse the ratio of DNT cells in peripheral blood with normal person and pancretic carcinoma patients and the ratio of DNT cells in tumor-infiltrating lymph nodes, the ratio of DNT cells pre-and postoperative to evaluate the immune funcion of pancreatic carcinoma patients,and to explore the role of DNT cells in anti-tumor immune response. At the same time,we detecte the IFN-γand IL-4 levels in pancreatic carcinoma patients and healthy control group ,to explore the relationship between the cytokin level and DNT cells.We also have further study,we isolate and culture the DNT cells and detect the cytokin level secreted by DNT cells,make the experimental basis for pancreatic carcinoma cell-killing effect by DNT cells.Objective:The study use the flow cytometry analyse the ratio of DNT cells in peripheral blood with normal person and pancretic carcinoma patients,ELISA methods to detect the IFN-γand IL-4 levels in pancreatic carcinoma patients and healthy control group, immunomagnetic beads methods to isolate the DNT cells,to analyse the relationship among DNT cells portion and clinical pathological stage of pancreatic carcinoma,lymph node metastasis,tumor size ,explore the relationship between IFN-γand IL-4 levels and DNT cells to evaluate the role of DNT cells in anti-tumor immune response. Methods:(1) To detect the ratio of DNT cells by flow cytometry: the ratio of DNT cells in peripheral blood with normal person and pancretic carcinoma patients and the ratio of DNT cells in tumor-infiltrating lymph nodes were detected by flow cytometry.(2) ELISA assay (antigen -antibody sandwiched): the IFN-γand IL-4 levels in heathy control group and in pancreatic carcinoma patients pre-and postoperative, the IFN-γand IL-4 levels secreted by DNT cells which we isolated and cultured.(3) Immunomagnetic beads methods: To isolate and culture the DNT cells by using immunomagnetic beads methods.Results:(1) The content of CD4~-CD8~- T cell in TCRαβ~+T cell from healthy person is (6.25±2.61)%,The content of CD4-CD8- T cell in TCRαβ~+T cell from pancreatic carcinoma patientst is (4.20±1.69)%,it can found significant deviation between them(p<0.01). The ratio of CD4~-CD8~- T cell in TCRαβ~+T cell from pancreatic carcinoma patients is(4.13±1.81)% before operation (4.20±1.69)%, the ratio of CD4~-CD8~- T cell in TCRαβ~+T cell from 20 pancreatic carcinoma patients is(5.90±1.27)% after operation, it can found significant deviation between them(P<0.01).(2)The content of DNT cells was associated with TNM grade and lymph node metastasis(P<0.01).But no relationship among hisological differentiation,age, gender and the tumor size( P>0.05).(3) The IL-4 level of pancreatic carcinoma patients is(86.29±23.29)pg/ml before operation; the IL-4 level of pancreatic carcinoma patients is(70.91±7.53)pg/ml after operation,it can found significant deviation between them(P<0.01),The IFN-γlevel of pancreatic carcinoma patients is(16.44±4.84)pg/ml before operation; the IFN-γlevel of pancreatic carcinoma patients is(17.45±5.21)pg/ml after operation,it can found no deviation between them( P>0.05). (4) TCRαβ~+ CD4~-CD8~- T cell can secret IFN-γ,don't seret IL-4.Conclusions:(1) The content of DNT cells was associated with TNM grade and lymph node metastasis.But no relationship among hisological differentiation,age, gender and the tumor size.(2) The CD4~-CD8~- T cells can secret IFN-γ,it can through cytokin to play antitumor activity. CD4~-CD8~- T cells may be a new way for tumor therapy.