| Objective The purpose is to study the FSTL1 stimulation DC of patients with NPC. After induction of maturation, culture with autologous lymphocyte, its supernatant was detected IL-12 and IL-4,and the expression of its total cellular protein was detected P-STAT4 and P-STAT6. Investigate the effects of FSTL1 on dendritic cells and lymphocyte immune function in patients with nasopharyngeal carcinoma, to explore its mechanism.Methods1 Collected peripheral blood 25ml by 33 nasopharyngeal carcinoma patients and 18 healthy persons, and extracted mononuclear cells (PBMC) out. Monocytes and lymphocytes are isolated by means of adherent. Lymphocytes were keeping constant state of growth culture if used in sometimes. Serum is stored at -20℃ for using in future. Culture of lymphocyte maintained its growth state for 8 days.2 Using tumor necrosis factor (1000U/ml), interleukin-4(500U/ml) and granulocyte-macrophage colony-stimulating factor (1000U/ml) stimulated cultured monocytes in vitro. The first five days DC is divided into experimental and control groups:the experimental group was added FSTL1, named FSTL1+DC group; control without FSTL1, named DC group. On eighth day the mature DC is collected.3 Mature DC present antigen peptides LMP2 to autologous lymphocytes, are mixed cultured. Cell supernatants is collected on day 12, total protein extracts after lysis of cells, FSTL1、IL-12 and IL-4 concentration of serum is dedected by ELISA, and the same to IL-12、IL-4 concentration of cell supernatants, in Western Blotting STAT4 and STAT6 protein expression of phosphorylation is detected.Results1 Successfully cultured mature dendritic cells,which the forms is diverse, the projections are not the same size, and the length is typical.2 Enzyme-linked immunosorbent assay results2.1 NPC patients and healthy human serum:① nasopharyngeal carcinoma patients and healthy person’s FSTL1、 IL-12 and IL-4 concentration of serum were compared, the difference was not statistically significant (P> 0.05).② FSTL1 were positively correlated with IL-4 and IL-12 showed a negative correlation, IL-4 and IL-12 were negative correlation, but the P values were greater than 0.05, not statistically significant.2.2 Patients with nasopharyngeal carcinoma and healthy person cell supernatants:① Nasopharyngeal Group:DC group and DC+FSTL1 group concentration of IL-12 secretion, the difference was not statistically significant (P>0.05); IL-4 concentration compared to two set of data significantly (P= 0.029), DC+FSTL1 group concentrations were higher than DC group. ② Healthy control group:DC group and DC+FSTL1 group concentration of IL-12 secretion, the difference was not statistically significant (P> 0.05); IL-4 concentration compare two sets of data were significantly different (P= 0.003), DC+FSTLl group was significantly higher than DC. ③ NPC patients and healthy people of DC+FSTL1 group compared:IL-12 concentration, the difference was not statistically significant (P> 0.05), IL-4 concentration, the difference was significant (P=0.001), healthy DC+FSTL1 group is much higher than in NPC group.④ NPC patients and healthy people compared with DC group:IL-4 IL-12 concentration and no statistically significant difference (P>0.05). ⑤ NPC cell supernatant experimental group Pearson correlation coefficient IL-12 and IL-4 concentration was between 0.21, P> 0.05, the difference was not statistically significant; the experimental group of normal cell supernatant IL-12 and IL-4 concentration Pearson correlation coefficient was 0.45, P>0.05, the difference was not statistically significant.3 Western Blot Result① nasopharyngeal carcinoma patients:DC group and DC+FSTL1 group, P-STAT4 semi-quantitative expression, the difference was significant (P= 0.000), DC+FSTL1 set much higher than the DC group; p-STAT6 semi-quantitative expression the difference was significant (P= 0.001), DC+ FSTL1 set much higher than the DC group. ② healthy control group:single DC group and DC+FSTL1 group, P-STAT4 semi-quantitative expression were significantly different (P=0.008); P-STAT6 semi-quantitative expression were significantly different (P= 0.003),DC+FSTLl group significantly higher than the DC group. ③ NPC patients and healthy people compared to DC+FSTL1 group, P-STAT4 semi-quantitative expression, significant differences (P= 0.001), far lower than in healthy patients with nasopharyngeal carcinoma; P-STAT6 semi-quantitative expression, the difference is very significant (P=0.005), significantly lower than in healthy patients with nasopharyngeal carcinoma. ④ NPC patients and healthy people compared with DC group, P-STAT4 semi-quantitative expression, the difference was not statistically significant (P=0.145); P-STAT6 semi-quantitative expression, significant differences (P= 0.005), healthy human DC group DC group was significantly higher than in NPC.Conclusion After FSTL1 stimulating by DC, presenting its activity increased, so that the lymphocytes secrete IL-4 increases, the impact STAT4 and STAT6 of phosphorylation levels, thereby immunity of lymphocyte is enhanced. |
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