Pilot Study On Change And Clinical Significance Of Peripheral CD4~+ T-cell Subsets After Radiofrequency Ablaiton In Patients With Lung Cancer

PurposePreliminary study on peripheral four distinct CD4~+T-cell subsets including Th1, Th2, Th17 and Treg in healthy individual. And preliminary study on the clinical significance and dynamic change of CD4~+T-cell subsets in lung cancer patients before and after the Radiofrequency ablation(RFA) therapy.Materials and MethodThe peripheral heparinized blood samples were obtained from30 healthy controls and 6 patients with lung cancer before and 0.5, 1, 2, 3 month after RFA. Percutaneous RFA were performed under computed tomographic (CT)-guided and intravenous anesthesia. Four distinct CD4~+T-cell subsetsTh1, Th2, Th17, Treg were assessed by flow cytometry. IFN-γ, IL-4, IL-17 were used as markers for the differentiation of Th1, Th2, Th17 subsets, respectively, using the whole blood intracellular staining assay to analyze the cytokines. These blood samples were added PMA, Ionomycin, Brefeldin, then incubated 6 hours. These three subsets were labeled fluorochrome-coupled antibodys CD3—PE-Cy5, CD8—APC-Cy7, IFN-γ—APC, IL-17—FITC, IL-4—PE, and detected by BD FACSCanto, analyzed by FACS Diva software. The characteristic of Treg subset was CD3~+CD4~+CD25~+foxp3~+, labeled fluorochrome-coupled antibodys CD3—APC, CD4—PE-Cy5, CD25—FITC, foxp3—PE, detected by BD FACSCanto, and analyzed by FACS Diva software. Data of healthy controls were presented as means±SD, analysised by Statistical software SPSS13.0.ResultThe levels of Th1, Th2, Th17 and Treg subsets in peripheral blood of healthy individuals were: 36.88%±9.43, 1.01%±0.56, 3.03%±1.00, 1.83%±1.07, respectively, and Th1/Th2 ratios was 53.09±38.35. Before RFA therapy, the level of Th1 cell and Th1/Th2 ratio of patients were lower than healthy control, while Th2, Th17, Treg cell subsets were higher. Half month after RFA therapy, The level of Thl cell increased, while Th2, Th17 and Treg cells decreased compared with preoperative levels. After then, The tendency fluctuate. However, the level of Thl cells was still higher than preoperative levels, and Th2, Th17 and Treg were still lower. Th1/Th2 ratio also increased 0.5 month after RFA therapy.ConclusionThe peripheral level of Th1cell in healthy individual was higher than Th2, Th17, Treg. In patients with lung cancer, the levels of Th1 decrease and Th2 increase, leading to the imbalance state of Thl/Th2, which impairs anti-tumor immunity. Th17 cell increase, enhancing angiogensis and lymphangiogenesis to promote lung cancer growth. And Treg cell increase, involved with development of tumor. RFA can correct the imbalance state of Thl/Th2, and induce a reduction of circulating Th17 and Treg, which may be used as indicator of clinical efficacy.