宫颈癌HPV 分型与Th17和Treg细胞的相关性研究*

目的：检测感染人乳头状瘤病毒（human papilloma virus,HPV ）患者的外周血中CD4+IL 17+Th17、CD4+CD25+Foxp 3+Treg 细胞,分析Th17/Treg比值,比较血清中IL- 17、IL- 10、TGF-β 、IL- 23细胞因子的表达差异。方法：收集2014年9 月至2016年6 月119 例新疆医科大学第一附属医院HPV 感染未行治疗患者的资料,分为宫颈癌（cervical cancer ,CC）组46例、宫颈上皮内瘤变（cervicalintraepithelial neoplasia ,CIN）Ⅱ～Ⅲ组43例、慢性宫颈炎（chronic cervicitis ,CCS）组30例,根据检测HPV 分型将患者HPV 感染分为高危型80例与低危型39例,流式细胞仪检测Treg 及Th17细胞,ELISA 检测相关细胞因子。结果：高危型HPV 感染患者中CC组感染率为87％（40/ 46）,显著高于CIN Ⅱ～Ⅲ组77%（33/ 43）及CCS 组23%（7/ 30）。 CC组外周血中Th17、Treg 细胞及血清中相关细胞因子均明显高于其他两组,且随着宫颈病变加重,Th17/Treg比值升高更加显著。高危型HPV 感染患者外周血中Th17和Treg 细胞高于低危型HPV 感染患者（P < 0.05）,Th17/Treg比值在高危型HPV 感染患者中显著升高。高危型HPV 感染患者血清中IL- 17、IL- 10、TGF-β 、IL- 23细胞因子的表达较低危型HPV 感染患者均显著增高（P < 0.05）。 结论：Treg 、Th17细胞在高危型HPV 感染患者中存在增高现象,Th17/Treg比值的升高可能直接参与宫颈HPV 的感染及免疫逃逸过程,在宫颈癌的发生发展中起着重要作用。     

Relative expression levels of Th1 and Th2 cytokine mRNA are independent prognostic factors in patients with ovarian cancer

The aim of the present study was to examine mRNA expression levels of Th1 (TNF-alpha , IFN-gamma, and IL-12p40) and Th2 (IL-6 and IL-10) cytokines for any association with clinicopathological characteristics of epithelial ovarian cancer. mRNA was isolated, and cDNA prepared from 40 samples of epithelial ovarian cancers. Expression level of each cytokine mRNA was examined by the real-time PCR technique (GAPDH gene, internal control). Expression ratio (target gene/GAPDH) was used to evaluate gene expression. Results were analyzed against clinical stage, histological grade, and histological type. Prognostic significance of expression levels of each combination of Th1/Th2 values was assessed. Tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) expression levels were significantly higher in serous adenocarcinoma than in non-serous adenocarcinoma (p<0.05), but with no difference between individual cytokine mRNA expression levels and clinical stage or histological grade. Log-rank testing showed that high TNF-alpha mRNA expression (p=0.033) and the diameter of largest residual lesion at initial surgery (p=0.012) significantly correlate with longer survival in advanced stage (II/III/IV) ovarian carcinomas. In examining all combinations of Th1/Th2 expression values, the most significant association was between high IFN-gamma.IL-12p40/IL-6 expression levels and better prognosis in advanced stage (II/III/IV) ovarian carcinomas (p=0.004). In multivariate analysis, high IFN-gamma.IL-12p40/IL-6 expression (p=0.009) and the diameter of residual lesion (p=0.011) remained significantly associated with survival, whereas high TNF-alpha expression lost significance. In conclusion, Th1 and Th2 cytokines might play an important role in regulating the immune reaction in epithelial ovarian cancer cells. IFN-gamma.IL-12p40/IL-6 expression may be a useful prognostic molecular marker for patients with advanced ovarian cancer.     

Decreased interleukin-1 receptor antagonist response following moderate exercise in patients with colorectal carcinoma after primary treatment

Clinical and experimental studies have shown that a moderately increased physical activity level may have beneficial effects in terms of exercise conditioning, resistance to infection and decreased relative risk of cancer. Modulation of the innate and adaptive components of the immune system with a shift of cytokines and their antagonists to a more pro- and less anti-inflammatory response was found to be a prominent feature in non-tumor patients and healthy volunteers. As quantitative data concerning the cytokine/antagonist response following exercise are not available for tumor patients, we compared the effects of a post-operative rehabilitation program with moderate exercise (ME) intensity (0.55-0.65 x maximal aerobic power) with a program with low exercise (LE) intensity (0.30-0.40 x maximal aerobic power) in patients with curatively treated colorectal carcinoma (UICC II and III) measuring pro- (IL-1beta, IL-6, tumour necrosis factor (TNF)) and anti-inflammatory cytokines (IL-1 receptor antagonist, sTNF receptors I and II). Twenty-three patients participated in this prospective trial, N = 13 in the ME group, N = 10 in the LE group. Exercise was performed daily 30-40 min for 2 weeks. Basal (circulating) and LPS-stimulated (phasic) cytokine and antagonist response was determined before exercise and after 1 and 2 weeks using appropriate ELISA tests. The LPS-stimulated interleukin-1 receptor antagonist (IL-1ra) response in the ME group gradually decreased from 31,532.6 (160.0-70,028.0) to 18,033.0 pg/ml (5040.0-52,570.0) after one and to 22,892.0 pg/ml (6376.0-34,726.0) after 2 weeks (P < 0.05) with a concomitant decrease of the corresponding IL-1ra/IL-6 and IL-1ra/IL-1beta ratio: 2.51-1.41 and 4.1-3.1, respectively. In contrast, in the LE group LPS-stimulated cytokines and antagonists did not significantly change during exercise. Circulating cytokines and antagonists remained unchanged in both groups. In providing quantitative data in patients with curatively-treated colorectal cancer, we demonstrated that a short-term rehabilitation program with moderate exercise leads to a decreased LPS-induced antagonist response with a shift to a more pro-inflammatory state (decreased antagonist/cytokine ratio). Whether this change of the phasic immune response to moderate exercise may be clinically beneficial (decreased rates of infection, relapses and/or second tumours) is possible, but has to be investigated in long-term studies.     

Abnormal levels of proinflammatory cytokines in serum and monocyte cultures from patients with chronic myeloid leukemia in different stages, and their role in prognosis.

Interleukin-1 (IL-1), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha), primarily monocyte-derived cytokines, form a group of proinflammatory cytokines with related and overlapping spectra of activities. The role of these cytokines in chronic myeloid leukemia (CML) has been investigated. A distinctive pattern of cytokine secretion has been found in chronic myeloid leukemia in chronic phase (CML-CP), in blastic crisis (CML-BC) and in normal subjects. Serum IL-6 levels in CML-CP and CML-BC were significantly raised compared with normal controls (p = 0.0026 for CML-CP and p = 0.0011 for CML-BC). IL-6 was significantly elevated in blastic crisis of CML (103.5 +/- 20.77 pg ml-1) compared with CML-CP (37.35 +/- 10.88 pg ml-1; p = 0.014). IL-6 serum levels were found to correlate significantly with peripheral blood monocyte counts and bone marrow blast and basophil counts. We have analysed monocyte/macrophage function with respect to their ability to produce IL-1, IL-6 and TNF-alpha, spontaneously as well as in response to LPS, in comparison with normal controls. A direct correlation of IL-6 levels in unstimulated and stimulated cultures with bone marrow blast and basophil counts has been observed. From these results it is inferred that the monocyte function is impaired in CML patients, and the cytokine secretion is deficient. Our limited data suggest that serum IL-6 levels may play an important role as a prognostic marker for CML.     

Evaluation of inflammatory cytokines in malignant and benign pleural effusions

We measured the levels of inflammatory cytokines interleukin-1alpha (IL-1alpha), interleukin-1beta (IL-1beta), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-alpha) in pleural effusions and serum in 65 consecutive patients: 32 with malignant pleural effusion (MPE) (group A), and 33 with inflammatory benign pleural effusion (BPE) (group B). Serum levels of 15 healthy individuals served as control. Concentrations of IL-1alpha were higher in serum compared to pleural fluid in both groups (47.1+/-33.9 vs. 25.9+/-1.7 fmol/ml, p<0.001, in group A; and 39.9+/-30.9 vs. 25.4+/-16.3 fmol/ml, p<0.02, in group B). Similarly, concentrations of IL-1beta and IL-2 were significantly higher in serum compared to pleural fluid in both groups. In contrast, IL-6, IL-8 and TNF-alpha were found at high concentration in MPE in comparison to serum IL-6: 171.8+/-60.4 vs. 7. 2+/-7 fmol/ml (p<0.001), IL-8: 1175.15+/-2385.6 vs. 285.2+/-187.2 pg/ml (p<0.05), TNF-alpha: 204.9+/-82.9 vs. 79.4+/-31.9 fmol/ml (p<0. 001). Similarly, pleural concentrations of IL-6, IL-8 and TNF-alpha were higher in BPE patients in comparison to serum IL-6: 124.3+/-56. 2 vs. 8.6+/-6.4 fmol/ml (p<0.001) IL-8: 2109.2+/-4121.5 vs. 291. 6+/-197.9 pg/ml (p<0.02), TNF-alpha: 183.8+/-28.2 vs. 86.2+/-23.9 fmol/ml (p<0.001). These data suggest that IL-6, IL-8 and TNF-alpha might be secreted locally at the site of active disease both in benign and malignant pleural effusions.     

Circulating cytokines in patients with metastatic cancer treated with recombinant interleukin 2 and lymphokine-activated killer cells

Treatment with recombinant interleukin 2 and lymphokine-activated killer cells (rIL-2/LAK) has produced a clinical antitumor effect in preliminary human trials. The cytokines gamma-interferon (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), and tumor necrosis factor beta (TNF-beta, lymphotoxin) have potent in vitro antitumor activity and some clinical toxicities similar to interleukin 2 (IL-2)/LAK. This study sought to determine whether these cytokines were detectable in sera of IL-2/LAK-treated patients. Ten patients were treated with a protocol of 5-day i.v. rIL-2 bolus priming (10(5) units/kg, every 8 h), followed by 5 daily phereses with harvested lymphocytes cultured in vitro to generate LAK, and 5 days of rIL-2 bolus with infusion of LAK cells. Five patients were treated with a protocol modified to a 3-day rIL-2 prime and 6-day continuous infusion rIL-2 (3 x 10(6) units/m2/day) with infusion of LAK cells. Serum specimens were obtained prior to and 0.5, 2, 3, and 5 h after IL-2 or LAK cell administrations. IFN-gamma was detected by enzyme-linked immunosorbent assay, TNF-alpha by WEHI 164 bioassay or enzyme-linked immunosorbent assay, and TNF-beta by WEHI 164 cell bioassay. During the prime, few patients manifested in vivo detectable serum cytokines: IFN-gamma, three of ten, 5-day prime (1.03 +/- 0.46 ng/ml), and zero of five, 3-day prime; TNF-alpha, one of ten, 5-day prime, and one of three, 3-day prime; TNF-beta, one of ten, 5-day prime. The supernatants of in vitro LAK generation cultures had detectable levels of cytokines at 24 h which increased progressively until culture harvest at Day 4 (IFN-gamma, 2.56 +/- 0.34 ng/ml; TNF-alpha, 356 +/- 110 pg/ml; TNF-beta, 8.2 +/- 4.4 units/ml). The highest levels of in vivo serum cytokines occurred following LAK cell infusion and were more often elevated in patients receiving rIL-2 by bolus than by continuous infusion: IFN-gamma, four of six bolus, zero of three continuous infusion; TNF-alpha, six of six bolus (maximum 679 pg/ml) versus two of three continuous infusion (maximum, 106 pg/ml). LAK cells in vitro responded with cytokine release on stimulation by tumor cell lines (IFN-gamma, 0.88 +/- 0.06 ng/ml; TNF-alpha, 426 +/- 16 pg/ml; TNF-beta, 0.64 +/- 0.06 units/ml). In summary, this preliminary study has detected circulating cytokines in sera of patients receiving IL-2/LAK therapy. The greatest cytokine elevations followed LAK cell infusion.(ABSTRACT TRUNCATED AT 400 WORDS)     

Serum cytokines in metastatic melanoma patients treated with an autologous tumor vaccine

Short-term autologous tumor vaccines were established and used to treat metastatic melanoma patients. Serum samples obtained prior to (week 0) and after three vaccinations (week 4) were assayed for interleukin (IL)-2, interferon (IFN)-gamma, IL-4, and IL-10. Results (mean +/- SD) for 30 patients who had matching serum samples obtained at weeks 0 and 4 were: week 0, IL-2, 122 +/- 320 pg/mL; IFN-gamma, 0.1 +/- 0.4 IU/mL; IL-4, 10.0 +/- 19 pg/mL; IL-10, 159 +/- 237 pg/mL; week 4: 119 +/- 308 for IL-2; 0.1 +/- 0.4 for IFN-gamma; 16 +/- 29 for IL-4, and 210 +/- 273 for IL-10. Medium conditioned by tumor cell lines demonstrated relatively low levels of secreted IL-10 (3.5 +/- 4.2 pg/106 cells/mL/96 hours), which would not account for the observed serum levels. In conclusion, the serum cytokine pattern from these patients suggests that the immune system is being modulated prior to and subsequent to vaccination.     

Measurement of serum cytokine levels in patients with myelodysplastic syndromes.

Sera of 25 healthy controls and 75 patients suffering from myelodysplastic syndromes (MDS) were investigated for serum concentration of interleukin-1 alpha (IL-1 alpha), IL-3, IL-6, granulocyte-colony-stimulating factor (G-CSF), granulocyte-macrophage-CSF (GM-CSF), erythropoietin (Epo), and tumor necrosis factor-alpha (TNF-alpha). According to French-American-British (FAB) classification, 21 refractory anemia (RA), seven refractory anemia with ring sideroblasts (RARS), 15 chronic myelomonocytic leukemia (CMML), 12 refractory anemia with excess of blasts (RAEB), and 20 RAEB in transformation (RAEBt) were examined. TNF-alpha levels were inversely correlated with lower levels of hemoglobin concentration (r = -0.31, p = 0.005), irrespective of the requirements for transfusion in anemic MDS patients. Significant differences in TNF-alpha levels between CMML (26.2 +/- 5.9 pg/ml) and the FAB subgroups (16.1 +/- 1.6 pg/ml) were detected. There was an overall inverse relationship between the level of erythropoietin and the degree of anemia, but a wide range of Epo response between patients with similar hemoglobin concentrations. Serum levels of IL-1 alpha and GM-CSF were undetected in most of the patients. In 57% of the samples there were detectable levels of G-CSF, without a correlation of the serum levels with blood cell counts, nor with any of the FAB subcategories. Overall, 29% and 25% of the patient sera exhibited elevated IL-3 and IL-6 levels, respectively. There was no correlation of the serum levels with any of the blood counts, other cytokines, nor FAB subcategories. In conclusion, simple negative feedback mechanism between a specific cytokine and the production of blood cells seems not to be the case in MDS, except for red cell production and erythropoietin concentration. Our data may suggest the involvement of TNF-alpha in the pathogenesis of anemia in MDS.     

Serum interleukin-2 (IL-2), soluble IL-2 receptors and tumor necrosis factor-alfa levels are significantly increased in acute myeloid leukemia patients

Serum interleukin-2 (IL-2), soluble IL-2 receptors (sIL-2R) and tumor necrosis factor-alfa (TNF-alpha) levels were determined in 66 previously untreated consecutive patients with acute myeloid leukemia (AML) and in 22 normal volunteers. The following mean (+/- SE) values were observed in patients and controls, respectively: 35 +/- 14.7 (range 0.5-500) and 0.7 +/- 0.02 (0.5-0.8 U/ml for IL-2 (p = 0.001); 1622 +/- 289 (110-10,600) and 422 +/- 30 (207-666) U/ml for sIL-2R (p = 0.0001); 1247 +/- 196 (218-4672) and 152 +/- 11 (75-308) pg/ml for TNF-alpha (p = 0.0001). With respect to the FAB classification system, we found a significantly different distribution of serum IL-2 mean values in distinct subcategories, i.e. 3.4 +/- 1.9 U/ml in M1-M2-M3 and 42.4 +/- 20.4 U/ml in M4-M5 subgroups, respectively (p = 0.01), whereas sIL-2R and TNF-alpha levels were 1144 +/- 322 U/ml and 1120 +/- 317 pg/ml in M1-M2-M3 patients and 1945 +/- 317 U/ml and 1270 +/- 259 pg/ml in the M4-M5 group. A significantly positive correlation between TNF-alpha and sIL-2R (r = 0.53; p = 0.002) was also detected in the M4-M5 group. Sixty-three out of 66 patients received an intensive chemotherapy program. Univariate analysis showed that age and sIL-2R greater than 2000 U/ml significantly affected both complete remission rate and overall survival, whereas by multivariate analysis, age was the only independent variable significantly influencing survival. These data confirm recent in vitro evidence suggesting the role of IL-2, sIL-2R, and TNF-alpha in the control of normal hematopoiesis and leukemogenesis. Since the availability of recombinant cytokines for clinical use in AML, it is crucial to understand their spectrum of interaction in order to select the appropriate combination for in vivo administration.     

Cellular immune profile of patients with advanced cancer before and after taxane treatment

The purpose of this study is to determine immune recovery and function after treatment with docetaxel or paclitaxel. Peripheral blood mononuclear cells were harvested before chemotherapy and at weekly times afterwards for cycle 1. Leukocyte subsets ICD45hiCD14lo polymorphonuclear neutrophils, CD45hiCD14hi monocytes, CD45hiCD14- lymphocytes, CD3+CD4/CD8+ T cells, CD3-CD19+ B cells, CD3-CD16/CD56+ natural killer (NK) cells], and circulating cytokine levels [tumor necrosis factor-alpha, gamma-interferon (gamma-IFN), and interleukins (IL-2, IL-10, IL-12)] were followed. In addition, T-cell mitogenic function, NK function, and lymphokine activated killer (LAK) function was assessed. Ten patients were entered in the trial. T-cell frequency, B-cell frequency, and CD4/CD8 ratio did not change. IL-10 serum levels significantly decreased in paclitaxel-treated patients (4.4+/-1.3 pg/ml at week 4 versus 7.8+/-2.1 pg/ml at baseline; p < 0.05). IL-2, IL-12, and gamma-IFN levels were not detectable. NK cytotoxic activity decreased in docetaxel-treated patients. LAK cell activity was not altered. Four patients achieved a partial or complete response. They demonstrated higher than normal CD4:CD8 T-cell ratios and an improved phytohemagglutinin stimulation index (SI = 2.5). In conclusion, our findings suggest that immune function was affected more significantly after docetaxel treatment. Investigational approaches, which enhance cellular immunity, may be of greater relevance after treatment with docetaxel. Additional studies monitoring NK function after chemotherapy are recommended.     

Serum interleukin-10 is an independent prognostic factor in advanced solid tumors

Interleukin (IL)-10 is a Th2 type pleiotropic cytokine that has been found to be produced at the tumor site and to be increased in sera of patients suffering from different types of cancer. IL-10 has been shown to hinder a number of immune functions, i.e., T lymphocyte proliferation, Th1 type cytokine production, antigen presentation, and lymphokine-activated killer cell cytotoxicity. To assess its prognostic value, we measured serum levels of IL-10 in 118 patients with advanced solid tumors before treatment, after completion of therapy, and during follow-up. Other prognostic variables, to which IL-10 results were compared, were analyzed as well. IL-10 serum levels were found significantly elevated in cancer patients with respect to healthy controls. Of interest, a significant decrease in IL-10 serum levels was observed in the responder group, whereas a significant increase was recorded in the non-responder group. Using univariate and multivariate analyses, a significant relationship was shown between IL-10 serum levels and both overall survival (OS) and time to treatment failure (TTF). Stepwise regression analysis selected IL-10 serum level, performance status (PS), and stage as the best association of variables with significant impact on OS and TTF. In conclusion, this study shows that IL-10 has an independent prognostic significance in patients with advanced solid tumors and may be useful for monitoring disease progression.     

胸腔镜手术与小切口开胸肺切除术后细胞因子反应的随机对照研究

背景与目的:细胞因子在诱导急性期炎症反应及免疫反应中起重要作用,能否通过微创入路手术减少细胞因子的释放,减轻术后急性期反应和免疫抑制是肿瘤外科治疗关心的问题.本研究希望通过前瞻性随机对照研究,比较电视辅助胸腔镜手术(video-assisted thoracoscopic surgery,VATS)与小切口开胸(minimal incision thoracotomy,MIT)肺切除术治疗临床早期非小细胞肺癌(non-small cell lung cancer,NSCLC),对比两种术式术后的细胞因子水平.方法:从2004年3月～2006年12月有47例CT提示为临床早期NSCLC、肿物直径≤6cm、符合入组条件的患者被随机分为VATS组(24例)和MIT组(23例),VATS组有2例患者因术中不能控制的出血和胸膜广泛粘连转为开胸手术而被剔除,MIT组1例因术中输血被剔除.使用流式微球分析技术(cytometric bead array,CBA)测定两组患者术前、术后4、24、48 h血浆中肿瘤坏死因子(tumor necrosis factor-α,TNF-α),白介素-2(interleukin-2,IL-2)、IL-4、IL-6和IL-10的浓度.结果:术后两组患者TNF-α、IL-2和IL-4的血浆浓度均较低,差异无统计学意义.术后4 h两组患者IL-6和IL-10均达峰值浓度.其中IL-6峰值浓度VATS组(91.0±63.9)ng/L,MIT组(84.2±53.1)ng/L,IL-10峰值浓度VATS组(12.6±8.1)ng/L,MIT组(16.3±11.2)ng/L,两组间差异无统计学意义(P=0.732和P=0.235).重复测量数据的方差分析显示两组间4个时间点IL-6和IL-10的浓度差异均无统计学意义(F=0.143,P=0.708和F=0.000,P=0.996).结论:胸腔镜手术和小切口开胸肺切除术后患者细胞因子反应无差异,其临床意义有待进一步研究.     

An analysis of cytokine status in the serum and effusions of patients with tuberculous and lung cancer

The present study was designed to ascertain whether or not the pleural effusion and serum cytokine levels (granulocyte-macrophage colony-stimulating factor [GM-CSF], interleukin-10 [IL-10], and interferon-gamma [IFN gamma]) in lung cancer patients differ from tuberculous (TB) pleural effusion, in which a strong cellular immune reaction is found; and, whether cytokine levels are a prognostic factor in lung cancer patients with malignant effusion. A total of 202 lung cancer patients with malignant pleural effusion and 26 patients with TB pleural effusion were studied consecutively between 1995 and 1998. Serum and effusion cytokine levels were analyzed with ELISA assays. The results showed that pleural effusion GM-CSF and IL-10 levels were significantly higher than serum levels in both cancer and TB patients. Pleural effusion IFN gamma levels were significantly higher than serum levels in TB patients. IFN gamma levels in both pleural effusion and serum were significantly higher in TB patients than in those with cancer. No significant difference was found, between TB and cancer patients, in the serum or pleural effusion levels of either IL-10 or GM-CSF. The ratio of pleural effusion IFN gamma to serum IFN gamma, effusion IFN gamma to effusion IL-10, and effusion IL-10 to serum IL-10, were all significantly higher in TB than in cancer patients, suggesting a higher cellular activity and T-helper 1 (Th1) reaction in TB pleural effusion than in malignant effusions, which were predominantly Th2 type. Survival analysis showed no significant difference in lung cancer patients with different levels of these cytokines. It was concluded that lung cancer patients with malignant pleural effusion had poorer immune profiles than those with TB pleurisy, both locally and systemically; and the cytokine profiles were not prognostic factors for lung cancer patients with malignant pleural effusion.     

Preoperative serum interleukin-18 level as a postoperative prognostic marker in patients with gastric carcinoma

BACKGROUND: Interleukin-18 (IL-18), a recently described cytokine produced mainly by macrophages, stimulates interferon-gamma (IFN-gamma) production by natural killer cells and T cells. Although it has been reported that serum IL-18 levels are higher in patients with advanced tuberculosis and acute graft-versus-host disease compared with normal controls, the authors found no reports regarding serum IL-18 levels in patients with malignant solid tumors. The purpose of this study was to determine serum IL-18 levels and their clinical significance in patients with gastric carcinoma. METHODS: Peripheral blood samples were obtained from 94 patients with gastric carcinoma who underwent curative surgery and from 50 healthy volunteers. The serum IL-18 level, the IFN-gamma, level, and the Helicobacter pylori (HP) serology status were determined in each sample with an enzyme-linked immunosorbent assay. RESULTS: The mean serum IL-18 level for all patients was significantly higher compared with the mean level in healthy volunteers (P < 0.01). IFN-gamma titers were below the level of detection in all samples tested. When the patients were subdivided into groups, it was found that the serum IL-18 level in patients with Stage II and III disease was significantly higher compared with the level found in healthy volunteers (P < 0.01). The serum IL-18 level decreased after patients underwent surgical resection. However, there was no significant difference in the serum IL-18 level between healthy controls and patients with Stage I or IV disease. Patients with IL-18 levels >or= 310 pg/mL (i.e., equal to or greater than the mean levels +/- 1 standard deviation in the healthy volunteers) experienced a significantly lower survival rate compared with patients who had IL-18 levels < 310 pg/mL after undergoing surgery (P < 0.05) despite a lack of any discernible difference in clinicopathologic factors between the two groups. The serum IL-18 level was identified as an independent postoperative prognostic factor in multivariate survival analysis using a Cox proportional hazards model (hazard ratio, 4.89; P = 0.01). There was no significant correlation between HP serology status and serum IL-18 levels. CONCLUSIONS: The preoperative serum IL-18 level may represent a significant postoperative prognostic determinant in patients with gastric carcinoma. Its function in the host immune system remains to be elucidated.     

宫颈癌患者血清Th1、Th2细胞因子表达水平及意义

目的 探讨宫颈癌患者血清Th1、Th2细胞因子表达水平及意义.方法 选取宫颈癌患者94例(观察组),同时选取健康女性90例作为对照组,检测两组患者干扰素-γ(IFN-γ)、白细胞介素-2(IL-2)、IL-4和IL-6.结果 观察组IFN-γ、IL-2、IL-4和IL-6水平分别为(52.13±9.55)pg/ml、(38.70±8.96)pg/ml、(27.61±6.22)pg/ml和(32.16±7.81)pg/ml,均高于对照组(P﹤0.05);Ⅲ~Ⅳ期患者IFN-γ、IL-2、IL-4和IL-6水平高于Ⅰ期和Ⅱ期患者(P﹤0.05),Ⅱ期患者IFN-γ、IL-2、IL-4和IL-6水平高于Ⅰ期患者(P﹤0.05);不同分化程度患者IFN-γ、IL-2、IL-4和IL-6水平比较差异无统计学意义(P﹥0.05),有无淋巴结转移患者IFN-γ、IL-2、IL-4和IL-6水平比较差异无统计学意义(P﹥0.05).结论 宫颈癌患者Th1/Th2细胞因子动态平衡紊乱,可能在肿瘤的发生发展中有一定作用.     

Cryosurgery for treatment of subcutaneously xenotransplanted tumors in rats and its effect on cellular immunity

Cryosurgery has shown encouraging therapeutic effects on some solid tumors but its effect on the cellular immunity remains unclear. We developed a subcutaneously xenotransplanted tumor model in SD rats to directly evaluate the immune response by detecting the serum cytokine levels, T-cell responses to tumor derived antigens, and the cytolytic activity of peripheral blood mononuclear cells against the W256 cancer line with apoptosis of cells being detected using TUNEL method. 66 SD rats were divided into 2 groups with group A having 36 rats and group B having 30 rats. 30 rats in group B were equally divided into 3 groups, tumor group B, cryosurgery group B and surgery group B, with 10 rats each. 36 rats in group A were equally divided into 2 groups, cryosurgery group A and tumor group A and treatment was done accordingly. The results showed that cryosurgery induced not only destruction of the tumor cells but also cell apoptosis around the cryosurgery foci. The apoptosis ratio reached the peak 12 h after cryosurgery, with an apoptosis rate of (68.28 ± 7.85)% .Compared with surgical resection that caused significant reduction in CD31 and CD41 cell percentages, cryosurgery resulted in significantly increased percentages of CD31 and CD41 cells (P < 0.05) with a relative increase of the CD41/CD81 cell ratio. However, sIL-2R level of peripheral blood of rats in cryosurgery group which decreased more rapidly than that in surgery group over time was significantly different 3 and 5 weeks after treatment compared to surgery group (P < 0.01). Moreover, cytotoxicity of mononuclear cell was significantly enhanced after cryosurgery, which is significantly higher in cryosurgery group (P = 0.05). These results demonstrate that in addition to tumor cell destruction, cryosurgery also results in enhanced cellular immunity and antitumor immune response of the rats with subcutaneously xenotransplanted tumor, suggesting the great potential of argon-helium cryosurgery in clinical management.     

Serum interleukin-10 levels in patients with advanced gastrointestinal malignancies

BACKGROUND: Interleukin-10 (IL-10) is a cytokine with immunosuppressive properties. In this study, the authors investigated the prognostic significance of IL-10 levels in the sera of 58 patients with advanced gastric or colorectal carcinoma. METHODS: IL-10 serum levels were measured before chemotherapy, on completion of chemotherapy, and at follow-up by means of a commercially available enzyme-linked immunoadsorbent assay kit. The results then were analyzed in comparison with other prognostic variables and a model predicting overall survival (OS) and time to disease progression (TTP) was generated. RESULTS: Elevated levels of serum IL-10 were found in carcinoma patients compared with healthy controls (19.6 +/- 6.8 pg/mL vs. 9.2 +/- 1.5 pg/mL; P < 0.0001), with those patients with metastatic disease showing significantly higher levels than patients with undisseminated disease (21.9 +/- 6. 7 pg/mL vs. 15.5 +/- 3.6 pg/mL; P = 0.0003). Retrospective analysis of prechemotherapy IL-10 serum levels showed a significant difference between responders and nonresponders (15.8 +/- 2.5 pg/mL vs. 21.6 +/- 7.6 pg/mL; P < 0.0001). Moreover, a further significant increase in IL-10 serum levels was observed in nonresponders at the end of therapy (21.6 +/- 7.6 pg/mL prechemotherapy vs. 31.3 +/- 11.6 pg/mL postchemotherapy; P < 0.0001) whereas no significant differences were observed in responders. Using univariate analysis, both OS and TTP were shown to be affected by the median pathologic levels of IL-10; multivariate analysis related to OS and TTP identified performance status and IL-10 serum level as the relevant prognostic factors, respectively. Finally, stepwise regression analysis identified IL-10 serum level and metastases as the prognostic factors related to both OS and TTP. CONCLUSIONS: The results of the current study show that measurement of pretreatment serum levels of IL-10 is of independent prognostic utility in patients with advanced gastrointestinal carcinoma and may be useful for the detection of disease progression.     

胃癌局部细胞因子表达谱分析

目的 探讨胃癌局部细胞因子表达的特点,为胃癌免疫治疗提供依据。方法 以相应非癌性黏膜组织为对照,采用高敏感放射性标记半定量RT－PCR技术,分析胃癌患者癌组织中纯化的CD4^ 、CD8^ T细胞亚群及上皮细胞的细胞因子表达谱。结果 胃癌组织与非癌性黏膜组织相比,CD4^ 和CD8^ T细胞亚群中均表现为Th2型细胞因子表达明显增加,Th1型细胞因子表达降低。其中癌组织CD8^ T细胞的IL－6、IL-8TNF－α的水平显著高于非癌性黏膜组织（P＝0．029,P＝0．022,P＝0．002）;胃癌组织的T细胞及癌性上皮细胞中细胞因子IL－10,TGFβ1和TNF－α的表达较非癌性黏膜组织增高。结论 胃癌局部Th1、Th2之间的平衡向Th2漂移,提示胃癌局部呈免疫抑制状态。CD8^ T细胞中细胞因子表达变化更为显著,提示CD8^ T细胞在胃癌局部免疫抑制发生中起主要作用。对不同细胞亚群细胞因子表达状况的了解有助于免疫治疗方案的设计。     

宫颈癌患者HPV感染状况及外周血Th1/Th2细胞因子变化研究

目的 探讨宫颈癌患者人乳头瘤病毒(HPV)感染状况及外周血Th1/Th2细胞因子变化,分析其在宫颈癌发生及发展中所起的可能机制,为宫颈癌的临床防治提供参考.方法 选取80例宫颈癌患者为研究对象(宫颈癌组),选取同期门诊体检的健康女性80例为对照(对照组),采用荧光定量聚合酶链反应(FQ-PCR)检测宫颈HPV-DNA,...