Effects of Shen-Fu Injection (参附注射液) on Apoptosis of Regulatory T Lymphocytes in Spleen during Post-Resuscitation Immune Dysfunction in A Porcine Model of Cardiac Arrest

Objective： To investigate whether Shen-Fu Injection（参附注射液, SFI） reduces post-resuscitation immune dysfunction in a porcine model of cardiac arrest by modulating apoptosis of regulatory T lymphocytes（Treg） in the spleen. Methods： After 8-min untreated ventricular fibrillation and 2-min basic life support, 24 pigs were divided into 3 groups with a random number table, i.e. SFI group, epinephrine（EP） group, and saline（SA） group（8 in each group）, which received central venous injection of SFI（1.0 m L/kg）, EP（0.02 mg/kg） and SA, respectively. The same procedure without CA initiation was achieved in the sham-operated（sham） group（n=6）. After successful return of spontaneous circulation（ROSC）, apoptosis rate of splenic Treg was detected by flow cytometry; and the m RNA expression of forkhead/winged helix transcription factor（Foxp3） of splenic Treg was detected by real time-polymerase chain reaction; and the levels of interleukin-4（IL-4） and interferon-γ（IFN-γ） in porcine splenic Treg were detected by using enzyme-linked immunosorbent assay（ELISA）. Results： Compared with the sham group, the apoptosis rate of Treg was significantly decreased, and the levels of Foxp3 m RNA expression, IFN-γ, IL-4 and IFN-γ/IL-4 were increased in the SA group（P〈0.05 or P〈0.01）. Compared with the EP and SA groups, SFI treatment increased the apoptosis rate of Treg and reduced the levels of Foxp3 m RNA expression, IFN-γ and IFN-γ/IL-4（P〈0.05）. Conclusions： SFI has significant effects in attenuating post-resuscitation immune dysfunction by modulating apoptosis of Treg in the spleen.

Changes of CD4^+CD25^+ Regulatory T Cells in Patients with Acute Coronary Syndrome and the Effects of Atorvastatin

The function of CD4＋CD25＋ regulatory T lymphocytes （Treg） in patients with acute coronary syndrome （ACS） and the effects of atorvastatin were investigated. Forty-eight patients with ACS were randomly divided into two groups： group C receiving conventional therapy （n=24）, and group C＋A receiving conventional therapy＋atorvastatin （10 mg/day, n=24）. T lymphocytes from ACS patients （before and 2 weeks after the treatment） or 18 healthy subjects were separated and the flow cytometry was used to measure the percentage of Treg. The inhibitory ability of Treg on effector T cells was determined by mixed lymphocyte reaction （MLR）. ELISA was used to measure the serum levels of cytokines （IL-10, TGF-β1 and IFN-γ） before and after treatment. The results showed that as compared with normal control group, Treg percentage was decreased significantly （P〈0.01）, the inhibitory ability of Treg on the T lymphocytes proliferation was reduced （P〈0.01）, IFN-γ levels were increased and IL-10 and TGF-β1 levels were lowered in ACS patients. After treatment with atorvastatin, Treg percentage and the inhibitory ability of Treg on T lymphocytes proliferation were significantly increased in ACS patients. Serum IFN-γ was decreased significantly, while IL-10 and TGF-β1 were elevated significantly as compared with the non-atorvastatin group. The number of Treg was positively correlated with serum TGF-β1, but negatively with serum IFN-γ and CRP. It was concluded that ACS was associated with decreased number and defected function of Treg, which may play an important role in initiating immune-inflammatory response in ACS. The inhibitory effects of atorvastatin on inflammation in ACS may be due to its beneficial effects on Treg and restoration of immune homeostasis.

INVESTIGATION OF REGULATORY T CELLS IN PATIENTS WITH NON-SMALL CELL LUNG CANCER

Objective To evaluate the prevalence of CD4 ^＋ CD25^high regulatory T cells （ Treg cells） in the peripheral blood mononuclear cells （PBMC） and tumor-infiltrating lymphocytes （TIL） of patients with non-small cell lung cancer （NSCLC） and to investigate immunosuppression to the progression of cancer. Methods Peripheral blood and tumor tissues were collected from 20 patients with NSCLC at the time of surgery. None of the patients received surgery, radiotherapy, chemotherapy, or other medical interventions before this study. Cancer stages of the patients were Ⅰ-Ⅲ A. Venous blood samples were obtained from 20 health donors. PBMC were isolated from blood samples by differential centrifugation over Ficoll-Hypaque. TILs were isolated from tumors by differential centrifugation over Ficoll-Hypaque and Percoll. Percentage of CD4^＋ CD25^highTr/CD4＋T in PBMC and TIL was assessed by the flow cytometry. Results The percentage of CD4^ ＋ CD25high Tr/ CD4 ^＋T in PBMC [ （4. 87 ± 1.22 ） % ] of NSCLC patients was significantly higher than that in healthy donors [ （ 2.36 ± 0. 72 ） % ] （ P 〈 0.01 ）. The percentage of CD4^＋ CD25^highTr/ CD4^＋ T in PBMC [ （5.40 ± 1.20） % ] of NSCLC patients in stage Ⅱ-Ⅲ A was significantly higher than that in stage Ⅰ [ （3. 87 ± 0. 22 ） % ] （ P 〈 0. 01 ）. The percentage of CD4 ＋ CD25hiShTr/ CD4 ＋ T in TIL[ （ 8. 66 ±0. 76） % ] of NSCLC patients in stage Ⅱ-Ⅲ A was significantly higher than that in stage Ⅰ [ （ 7. 04 ± 0. 80） % ] （ P 〈 0. 01 ）. Conclusion The prevalence of CD4 ^＋ CD25^highTreg cells in PBMC and TIL of NSCLC patients was significantly higher than that in healthy donors. These Treg cells may be preventing appropriate antitumor immune responses. The population of CD4^ ＋ CD25^highTreg cells in PBMC and TILs of NSCLC patients with Ⅱ-Ⅲ A stage was significantly higher than that of NSCLC patients with Ⅰ stage. These Treg cells may facilitate development of tumors.

Immune response activation following hyperthermic intraperitoneal chemotherapy for peritoneal metastases:A pilot study

BACKGROUND Hyperthermic intraperitoneal chemotherapy(HIPEC)for peritoneal metastases(PM)is considered to be feasible,safe and to improve survival.AIM To investigate whether an immune response is activated following HIPEC for PM.METHODS Six patients were enrolled in this study.Peripheral blood samples were obtained from each patient prior to(day 0)and post-procedure(day 30),and used to evaluate the number of CD3+total,CD3+/CD4+T-Helper,CD3+/CD8+cytotoxic T,CD3+/CD56+natural killer and CD19+B lymphocyte numbers,and CD4+:CD8+T lymphocyte ratios.RESULTS The total numbers of CD3+,CD3+/CD4+T-Helper,CD3+/CD8+cytotoxic T,CD3+/CD56+natural killer and CD19+B lymphocytes,and CD4+:CD8+lymphocyte ratios were increased in all but one patient 30 d following the cytoreductive surgery-HIPEC procedure,and these increases were significant(P≤0.05)for CD3+/CD4+T Helper and CD3+/CD8+cytotoxic T lymphocyte numbers.CONCLUSION This report provides the first evidence that HIPEC exhibits immunomodulating activity in PM patients,resulting in generalized activation of the adaptive immune response.Moreover,the majority of lymphocyte populations increased following HIPEC and continued to be elevated several weeks following the procedure,consistent with a potential authentic immunomodulating effect rather than a normal inflammatory response,to be fully characterised in future studies.

Targeting Foxp3^＋ regulatory T cells-related immunosuppression for cancer immunotherapy

Objective To review the current research into Foxp3^＋ regulatory T cells （Treg） cell surface molecules, plasticity of Treg cells and mechanisms of Treg cell suppression and to explore the possibilities to interfere in Treg cell suppression of anti-tumor immunity. Data sources A literature search of all English articles was performed on the online electronic PubMed database dated 1995 to 2010. The keywords searched included： CD4^＋CD25^＋Foxp3^＋ regulatory T lymphocytes, cancer, and immunotherapy. After finding relevant articles within these search limits, a manual search was conducted through the references from these articles. Study selection Articles regarding the role of Treg cells in tumor immunity and the utility of Treg cells in tumor immunotherapy. Results The results show that significant numbers of Treg cells are found in many tumors and it has been shown that the number of tumor infiltrating Treg cells correlates with adverse clinic outcomes. Treg cells are emerging as a key component of acquired tolerance to tumors. Conclusions Several mechanisms of immunosuppression can be mediated by Treg cell function. Distinct immunosuppressive molecules expressed by Treg cells or diverse molecules related to Treg induction or migration represent potential drug targets for caner immunotherapy.