| Objective: CD4+CD25+regulatory T cell (Tr), a kind of immunosuppressor cells, may inhibit the development and activation of tumor-specific cells which can recognize tumor, and play an important role in tumor immune-tolerance and immune-escape. Our study was designed to evaluate the character of Tr in the peripheral blood of the patients with NHL, and try to find the difference of the Tr level between the patients of initial-therapy, the patients who had got CR after chemotherapy and the normal control, hoping to disclose immunosuppressing status in the NHL patients, so as to investigate the relationship between immunosuppression and the development of the disease, the effect of chemotherapy on the immune system, and also the effect of immuno-suppression on the disease relapse. All of these will provide theory reference for further immunotherapy of the cancer.Methods: Three groups were included, that is initial-therapy group with 30 patients without chemotherapy, CR/PR group with 30 patients after 4-8 cycles of chemotherapy and 15 normal control. The normal control are the out-patient for healthy examination, with neoplasm or infection excluded. The patients were selected from the inpatient of the Fourth Hospital of the He bei Medical University from Oct 2005 to Oct 2006, diagnosed by pathology. 1ml blood was sampled from the patients in the morning with EDTA-K2 anticoagulated. Flow cytometry was applied to detect the membrane CD3,CD4 and CD25 of the lymphocyte in the peripheral blood , the detail is as follow: first to take the antibody of CD3,CD4 and CD25 20μl respectively into the EDTA-K2 anticoagulated blood of 0.1ml , then to incubat in the room temperature for 30 minutes , and then add red blood cell lysate, Red blood cell was to be disposed by rupture of membrane and then fixed, the result to be analyzed by computer. The detection condition and parameter of flow cytometry is as follow: The flow cytometer of Epics-XLⅡtype from BECKMAN COULTER corporation in American was used and the excitation light source was 15mW by argon ion laser, and the wave length was 488nm; Expo 32 ADC was applied to analyze immunofluorescence data, Muticycle AV was applied to analyze DNA cell cycle fit. The flow-check TM Fluorpheres(10μm) fluorescence microballoons (REF 6605359. Beckman Coulter, Inc. Fullerton, CA 92835) was used as standard sample to adjust CV value within 2%.Results1 The proportion of the sub-group of lymphocyte in the peripheral blood of normal control: The proportion of CD3+,CD4+,CD25+,CD4+CD25+ lymphocyte in the peripheral blood of normal control is (70.51±15.35) %, (37.41±14.23)%,(2.21±1.01)%,(1.96±0.92)% respectively , agreed with that of other literature.2 The proportion of CD3+ lymphocyte in the peripheral blood of initial-therapy group is (68.65±9.44)%, lower than that of the normal control (70.51±15.35)% slightly without significance(P>0.05), and so did the CD4+ T cells, with the proportion of (25.99±8.56) % versus (37.41±14.23) % respectively but with significance(P<0.05); The proportion of the CD25+ and CD4+CD25+ Tr cells in initial treatment patients was significantly higher than that of the normal control, that is (7.64±7.41) % versus (2.21±1.01) % (P<0.01) and (6.26±3.91) % versus (1.96±0.92) % respectively (P<0.01).3 We did not find any correlation between the proportion of CD4+CD25+ Tr in the peripheral blood and the age, sex, PS state, clinical stage or pathology category of the patients in the initial-therapy group.4 The proportion of CD3+ T cells in remission group after chemotherapy is (69.75±9.48) % , it was higher than that of the initial-therapy group but lower than that of the normal control without significance (P>0.05). The proportion of CD4+ T cells in remission group after chemotherapy is (34.66±6.61) % , it was much higher than that of the initial-therapy group with significance (P< 0.05), and lower than that of the normal control slightly without significance (P>0.05); the proportion of the CD25+ in remission group was lower than that of the initial-therapy group slightly without significance (P>0.05), while much higher than that of the normal control with significance (P<0.01); the proportion of CD4+CD25+ Tr cells in remission group was ( 3.25±2. 94) %,significantly lower than that of the initial-therapy group with significance (P<0.05), and higher than that of the normal control with significance (P<0.05). 5 we did not find the difference of the proportion of CD4+CD25+ Tr between the CR group and the PR group.Conclusions1 In our study, the proportion of CD3+ T cells in peripheral blood of the patients with NHL before chemotherapy was normal and that of CD4+ T was significantly lower than that of the normal control, meanwhile, the proportion of CD4+CD25+ Tr is much higher, thus the patients with NHL exist the cellular immune suppression.2 In our study, we found that the CD4+CD25+ Tr in the remission group is much lower than that of the initial-therapy group, and much higher than normal group. While the level of Tr in the remission group was still in a higher level than the normal control, it suggest that the patients in remission were still in a immunosuppressive status, and it is necessary to take some intervention.3 In our study, we did not find the difference of the proportion of CD4+CD25+ Tr between the CR group and the PR group. The increased level of the CD4+CD25+ Tr in the CR may be one of the main relapsing causes. As for the patients with CR, we are observing the rate of relapse in different level of Tr, while it is too early to make anyconclusion.4 In our study, the proportion of the CD4+CD25+ Tr was in a high level in the initial therapy group and the remission group , agreed with the result of clinical observation, thus it suggested that the proportion of CD4+CD25+ Tr may be a useful index to reflect the cellular immune function. |
PDF Full Text Request