Effects Of IL-21, IL-2 On The Expression Of Cytokine In Peripheral Blood T Lymphocytes Cells Of The Patients With Oral Cancer And Clinical Significance

Objective:At present, the treatment of oral cancer is still mainly based on surgery, supplemented by local radiotherapy and systemic chemotherapy combined treatment, but the dysfunction and abnormal appearance of the patient caused by surgery can not be beared, while radiotherapy and chemotherapy on patients also cause serious side effects. Therefore, the study for the immunotherapy of patients with oral cancer has far-reaching clinical value. At present IL-2, CSF, IFN and TNF-αare applied to other tumor therapy. IL-21 is one of the IL-2 family, the new member in the structure and function is similar to IL-2. It has ability of long-lasting potentiation the activation of T cells, NK cell, and with little toxic side-effects. Thus, IL-21 have become an increasing concern, its most promising application prospects is the biological treatment of tumor area. In our study, the aim is to observe the expression of IL-2+, IFN-γ+, CD4+/IL-2+, CD4+/IFN-γ+ of T lymphocytes and time correlation by stimulating peripheral blood in oral cancer patients with IL-21 alone or in combination IL-2, to explore the impact of IL-21 alone or combined application with IL-2 in anti-tumor immune function of oral cancer patients and provide a theoretical basis for immunization.Methods: Taking early morning fasting venous heparin anticoagulation 3ml, and RPMI 1640 culture medium containing 10% inactivated newborn calf serum, 100U/ml penicillin and 100ug/ml streptomycin mix 1:1,and then put it in 24 micro-pore culture plates. There are two groups, one is blank control group and the other one is test group. The control group has no any cytokine and the test group according to their different cytokines added it were divided into A, B, C group. A: IL-2 group, B: IL-21 group, C: IL-2 + IL-21 group. (Amount of cytokines: IL-2 1000U/ml, IL-21 15ng/ml). They are cultivated in 37℃, 5%CO2, saturated humidity.Measure IL-2+, IFN-γ+,CD4+/IL-2+, CD4+/IFN-γ+ T-lymphocyte percentage in whole blood cultured cells by flow cytometry in 24 hour and 48 hour in order to observe the changes of its secretion and time-dependent.Results:1 The changes of cytokines in T-lymphocyte1.1 In 24-hours, IL-2+ T-lymphocyte expression of control group is 1.2313+0.1703%, while the A, B, C groups respectively are 3.0590 +0.3897%, 3.2043+0.5345%, 3.1740+0.4612%. In 48 hours, when the control group is 1.2486+0.0964% , the test group are 6.2083+1.2570%,4.7463+0.6297%,5.3133+1.0607%.In the two detection time point, the result of test groups are higher than the control group. IL-2+ T-lymphocyte ratio is significantly increased(P<0.05). There is no significant difference among the A, B and C groups (P> 0.05).1.2 In 24-hour, IFN-γ+ T-lymphocyte expression of control group is 1.0033+0.0919%, while the A, B, C three groups are 2.6447+0.3484%, 2.5103+0.2728%, 2.5390+0.2691%. In 48 hours, when the control group is 1.0128+0.1025%, the test groups are 5.9977+0.9969%, 5.6327+1.121%, 5.5280+0.8846%. In the two detection time point, the result of test groups are higher than the control group. IFN-γ+ T-lymphocyte ratio is significantly increased(P <0.05). There is no significant difference among the A, B and C groups (P> 0.05).1.3 In 24-hour, CD4+/IL-2+ T-lymphocyte expression of control group is 0.6670+0.1096%, while the A, B, C three groups are 2.2660+0.3767%, 2.3727+0.3341%, 2.4577+0.3828%. In 48 hours, when the control group is 0.6738+0.0917%, the test groups are 5.5490+1.0036%, 4.7463+ 0.6297%, 5.2780+0.8209%. In the two detection time point, the result of test groups are higher than the control group. CD4+/IL-2+ T-lymphocyte ratio is significantly increased(P<0.05). There is no significant difference among the A, B and C groups (P> 0.05).1.4 In 24-hour, CD4+/IFN-γ+ T-lymphocyte expression of control group is 0.5033+0.0846%, while the A, B, C three groups are 1.4853+0.2079%, 1.4027+0.1747%, 1.6113+0.2101%. In 48 hours, when the control group is 0.5128+0.0735% , the test groups are 4.7390+0.5536%, 5.0293%+0.7189,5.2777+0.8083%. In the two detection time point, the result of test groups are higher than the control group. CD4+/IFN-γ+ T-lymphocyte ratio is significantly increased(P<0.05), There is no significant difference among the A, B and C groups (P> 0.05).2 Comparation of the test results of the test groups in different time2.1 IL-2+: The expression of IL-2+ is significantly increased after cultured(P <0.05). Comparing the two time points, the IL-2+ expression of A group cultured for 48 hour is higher than the expression cultured for 24 hour, with statistical significance (P <0.05). The IL-2+ expression of B group and C group cultured for 48 hours is higher than the expression cultured for 24 hours, but no statistical significance (P> 0.05).2.2 IFN-γ+: The expression of IFN-γ+ is significantly increased after cultured(P<0.05). Comparing the two time points, the IFN-γ+expression of A, B, C three groups cultured for 48 hours is higher than the expression cultured for 24 hours, with statistical significance (P <0.05).2.3 CD4+/IL-2+ : The expression of CD4+/IL-2+ is significantly increased after cultured(P<0.05). Comparing the two time points, the CD4+/IL-2+ expression of A, B, C three groups cultured for 48 hours is higher than the expression cultured for 24 hours, with statistical significance (P <0.05).2.4 CD4+/IFN-γ+: The expression of CD4+/IFN-γ+ is significantly increased after cultured(P<0.05). Comparing the two time points, the CD4+/IFN-γ+ expression of A, B, C three groups cultured for 48 hours is higher than the expression cultured for 24 hours, with statistical significance (P <0.05).3 The correlation of between the anti-tumor immune function enhanced by IL-21, IL-2 alone or in combination stimulated and lymph node metastasis and clinical stages3.1 IL-2 stimulation3.1.1 The relationship with lymph node metastasis: the enhancement effect of IL-2+, IFN-γ+, CD4+/ IL-2+, CD4+/ IFN-γ+ in no lymph node metastasis group cultured for 48 hours is higher than lymph node metastasis group. CD4+/IFN-γ+ enhancement effect was significant (P <0.05), IL-2+, IFN-γ+, CD4+/ IL-2+ enhancement effect was not significant (P> 0.05 ).3.1.2 The relationship with clinical stage: the enhancement effect of the IL-2+, IFN-γ+, CD4+/IL-2+, CD4+/IFN-γ+ in clinical stageⅠ,Ⅱcultured for 48 hours is higher than the clinical stageⅢ,Ⅳ. IL-2+, IFN-γ+, CD4+/IFN-γ+enhancement effect was significant (P <0.05), CD4+/IL-2+ enhancement effect was not significant (P> 0.05 ).3.2 IL-21 stimulation3.2.1 The relationship with lymph node metastasis: the enhancement effect of IL-2+, IFN-γ+, CD4+/ IL-2+, CD4+/ IFN-γ+ in no lymph node metastasis group cultured for 48 hours is higher than lymph node metastasis group. IL-2+, IFN-γ+ enhancement effect was significant (P <0.05), CD4+/IL-2+, CD4+/IFN-γ+ enhancement effect was not statistically significant (P> 0.05).3.2.2 The relationship with the clinical stage: the enhancement effect of the IL-2+, IFN-γ+, CD4+/IL-2+, CD4+/IFN-γ+ in clinical stageⅠ,Ⅱcultured for 48 hours is higher than the clinical stageⅢ,Ⅳ. IL-2+, IFN-γ+, CD4+/IFN-γ+ enhancement effect was significant (P <0.05), CD4+/IL-2+ enhancement effect was not statistically significant (P> 0.05).3.3 IL-2 + IL-21 stimulation3.3.1 The relationship with lymph node metastasis: the enhancement effect of IL-2+, IFN-γ+, CD4+/ IL-2+, CD4+/ IFN-γ+ in no lymph node metastasis group cultured for 48 hours is higher than lymph node metastasis group. CD4+/ IFN-γ+ enhancement effect was significant (P <0.05), IL-2 +, IFN-γ+, CD4 +/ IL-2+ enhancement effect was not statistically significant (P> 0.05).3.3.2 The relationship with the clinical stage: the enhancement effect of the IL-2+, IFN-γ+, CD4+/IL-2+, CD4+/IFN-γ+ in clinical stageⅠ,Ⅱcultured for 48 hours is higher than the clinical stageⅢ,Ⅳ. IL-2+, IFN-γ+ enhancement effect was significant (P <0.05), CD4+/ IFN-γ+, CD4+/ IL-2+ enhancement effect was not statistically significant (P> 0.05).Conclusion:1 The single application of IL-21, IL-2 or both in combination, can enhance the expression of IL-2+, IFN-γ+, CD4+/IL-2+, CD4+/IFN-γ+ T-lymphocytes. It suggests that application of IL-21 may enhance the body's anti-tumor immune capability.2 The secretion levels of T lymphocytes cytokine of oral cancer patients affected by IL-21 and IL-2 did not differ significantly, suggesting that IL-21 has potential as a substitute of IL-2.3 IL-21 and IL-2 has no obvious synergistic reaction in enhancing the body's state of anti-tumor immunity.4 The oral cancer patients IL-2,IFN-γ+,CD4+/IL-2+,CD4+/IFN-γ+ T lymphocytes expression stimulated by IL-2 or IL-21 existences time correlation, suggesting that extending the time of IL-21 on the body can enhance the anti-tumor immune function.5 The anti-tumor immune function enhanced by IL-21, IL-2 alone or in combination stimulated and lymph node metastasis and clinical stages are related, suggesting that the anti-tumor immune function of patients with early stage oral cancer may be more easily enhanced than the later ones.