胰腺癌患者外周血CD4-CD8-T细胞与IL-4、IFN-γ的关系

目的 研究胰腺癌患者外周血CD4-CD8-T细胞比例与IL-4、IFN-γ含量变化及其相互关系.方法 分别应用流式细胞仪及ELISA法检测25例胰腺癌患者和45名健康人外周血中CD4-CD8-T细胞比例和IL-4、IFN-γ的含量.结果 25例胰腺癌患者CD4-CD8-T细胞占CD3+T细胞的比例为(4.2±1.7)%,45名健康人CD4-CD8-T细胞占CD3+T细胞的比例为(6.3±2.6)%,两组之间差异有统计学意义(P＜0.01).胰腺癌患者外周血中IL-4含量为(86.3±23.3)fg/L,健康对照组外周血中IL-4含量为(56.2±9.2)fg/L,两组之间差异有统计学意义(P＜0.01).胰腺癌患者外周血中IFN-γ含量为(16.4±4.8)fg/L,健康对照组外周血中IFN-γ含量为(27.4±3.8)fg/L,两组之间差异有统计学意义(P＜0.01).胰腺癌患者手术后CD4-CD8-T细胞比例显著高于术前(P＜0.01).胰腺癌患者外周血CD4-CD8-T细胞比例与IL-4含量呈负相关,与IFN-γ含量呈正相关;胰腺癌患者CD4-CD8-T细胞比例、IL-4含量与肿瘤临床分期有关(P＜0.05),与肿瘤分化程度均无关(P＞0.05),IFN-γ含量与肿瘤临床分期和肿瘤分化程度均无关(P＞0.05).结论 CD4-CD8-T细胞在胰腺癌患者外周血中比例降低,CD4-CD8-T细胞可能通过影响IFN-γ的分泌参与胰腺癌的发生和发展.     

胰腺癌患者手术前后CD4-CD8-T细胞比例与IL-4,IFN-γ含量的关系

目的研究胰腺癌患者手术前后外周血CD4-CD8-T细胞比例与IL-4、IFN-γ含量变化及其相互关系。方法分别应用流式细胞仪及ELISA法检测胰腺癌患者手术前后外周血中CD4-CD8-T细胞比例与IL-4、IFN-γ含量。结果20例胰腺癌患者术前CD4-CD8-T细胞占CD3 T细胞的比例为(4.13%±1.81%),术后CD4-CD8-T细胞占CD3 T细胞的比例为(6.11%±1.40%),两组之间差异有统计学意义(P<0.01)。胰腺癌患者术前外周血中IL-4含量为(90.29±24.99)pg/ml,术后外周血中IL-4含量为(71.43±8.39)pg/ml,两组之间差异有统计学意义(P<0.01);术前外周血中IFN-γ含量为(15.79±5.90)pg/ml,术后外周血中IFN-γ含量为(20.08±6.60)pg/ml,两组之间差异有统计学意义(P<0.01)。结论胰腺癌患者手术前后CD4-CD8-T细胞比例变化可能影响IL-4、IFN-γ含量的变化,胰腺癌患者手术前后测定CD4-CD8-T细胞比例、IL-4、IFN-γ含量对判断机体的免疫水平具有一定的参考价值。     

胰腺癌患者外周血CD4-CD8-T细胞的检测及其临床意义

目的 研究胰腺癌患者外周血CD4-CD8-T细胞含量,并探讨其含量与肿瘤的临床病理类型及临床分期的相关性.方法 应用流式细胞仪分析胰腺癌患者和健康对照组外周血中CD4-CD8-T细胞含量.结果 20例胰腺癌患者CD4-CD8-T细胞占CD3+T细胞的比例为(4.13±1.81)%;20例健康对照组CD4-CD8-T细胞占CD3+T细胞的比例为(6.39±1.83)%,两组之间差异有统计学意义(P＜0.01).高、中分化的胰腺癌患者CD4-CD8-T细胞占CD3+T细胞的比例为(4.41±1.66)%;低分化胰腺癌患者CD4-CD8-T细胞占CD3+T细胞比例为(4.18±2.32)%,两组之间差异无统计学意义(P＞0.05).Ⅲ、Ⅳ期胰腺癌患者CD4-CD8-T细胞占CD3+T细胞的比例为(2.96±1.50)%;Ⅰ、Ⅱ期胰腺癌患者CD4-CD8-T细胞占CD3+T细胞的比例为(5.09±1.50)%,两组之间差异有统计学意义(P＜0.01).结论 CD4-CD8-T细胞在胰腺癌中低表达,可能促进胰腺癌进一步发生发展.     

自体血回输对全髋关节置换术患者免疫功能的影响

目的 比较回收式自体血回输和异体血输血对全髋关节置换手术患者免疫功能的影响. 方法 选择50例全髋关节置换手术,随机数字表法分为自体血回输组(A组)和异体血输血组(B组),每组25例.术中根据血容量丢失情况分别用自体血回输及异体血输血,于麻醉前、输血后第2天和输血后第5天采用流式细胞仪测定血浆CD4+T淋巴细胞(CD4+)、CD8+T淋巴细胞(CDx+)、自然杀伤细胞(natural killer cell,NK)的比例以及白细胞介素(interleukin,IL)-2(IL-2)和白细胞介素-6(IL-6)的值.结果 B组输血后CD4+、CD8+、NK细胞、IL-2的值在第2天[(35±6)、(22±6)、(7±3)％、(523±407) ng/L]和第5天[(35±6)、(26±8)、(6±4)％、(442±376) ng/L]均低于术前[(40±8)、(28±9)、(9±4)％、(839±472)ng/L] (P＜0.05);A组输血后CD4+、CD8+的值在第5天[(39±8)、(27±9) ng/L]、NK细胞、IL-2的值在第2天[(8±4)％、(807±534) ng/L]和第5天[(8±4)％、(821±437) ng/L]均较术前有所下降,但差异无统计学意义(P＞0.05);IL-6的值在第2天[(3198±698) ng/L]和第5天[(3076±703) ng/L]均较术前[(2593±784) ng/L]有所升高(P＜0.05). 结论 自体血回输对全髋关节置换手术患者细胞和体液免疫功能均无明显抑制作用,是安全、可靠的血液保护方式.     

白细胞介素-15对小鼠胃癌的治疗作用

目的 建立小鼠胃癌模型,检测荷瘤状态对小鼠免疫细胞的影响,同时给予白细胞介素-15(IL-15)免疫基因治疗,以检测IL-15对胃癌的预防及治疗效果.方法 采用多因素联合攻击法诱导小鼠胃癌,同时采用IL-15表达质粒载体进行免疫基因治疗.20周后取胃标本行病理学检查,并取血及脾细胞检测T细胞亚群及CD4+ CD25+调节性T细胞(Treg).同时进行自然杀伤(NK)细胞的细胞毒性试验,通过统计学方法分析结果.结果 多因素联合攻击法诱导胃癌发生率为37.5％.诱癌组小鼠血清IL-15水平(9.20±2.92) ng/L明显降低(P＜0.05),且其脾NK细胞杀伤活性(216.91±117.80) U/L明显降低(P＜0.05).诱癌组小鼠外周血Treg水平(8.07±6.62)％明显升高(P＜0.05),而对照组小鼠脾细胞Treg水平(4.40±3.34)％明显降低(P＜0.05).各组小鼠之间外周血T细胞亚群变化差异无统计学意义(P＞0.05).诱癌组小鼠脾细胞CD3+T细胞水平(78.31±29.79)％明显升高(P＜0.05),而CD4+T细胞(19.98±5.77)％及CD8+T细胞水平(10.15±1.72)％明显降低(P＜0.05).结论 多因素联合攻击法为小鼠胃癌模型建立提供了良好的模型.小鼠荷瘤状态下免疫功能下降,通过IL-15免疫基因治疗可提高机免疫功能,起到抗肿瘤的效果.     

甲状腺肿瘤患者中辅助性T细胞22及白细胞介素-22的变化及意义

目的 检测甲状腺肿瘤患者外周血及肿瘤组织中辅助性T细胞22 (Th22)及白细胞介素(IL)-22的表达水平,探讨其与患者临床病理特征的关系.方法 选择56例原发甲状腺肿瘤患者和30例健康志愿者,采用流式细胞仪检测外周血Th22细胞水平,酶联免疫吸附试验法(ELISA)检测血清中IL-22表达水平,应用免疫组织化学双染色分析正常组织及肿瘤组织中Th22阳性细胞的浸润程度,分析这些参数与临床病理特征间的关系.结果 甲状腺肿瘤患者外周血中Th22细胞及IL-22水平分别为(1.74±0.27)％和(162.7±32.2) ng/L,显著高于对照组[(0.57±0.14)％,t=15.72,P＜0.01;(64.2 ±20.7) ng/L,t=11.31,P＜0.01];甲状腺癌患者的Th22细胞比例及IL-22水平明显高于甲状腺瘤患者[(2.21±0.34)％比(1.39±0.22)％,t=9.14,P＜0.01;(199.1±34.1)ng/L比(129.8±31.3) ng/L,t=6.79,P＜0.01].甲状腺肿瘤患者组织中Th22阳性细胞浸润率为56.25％,而对应正常组织为8.33％,差异有统计学意义(x2=8.167,P＜0.01);甲状腺癌组织中Th22细胞浸润率为78.6％,甲状腺瘤组织为38.9％,差异有统计学意义(x2=5.039,P＜0.05).外周血Th22细胞比例、IL-22的表达水平和肿瘤组织中Th22阳性细胞浸润程度与临床分期、淋巴结转明显相关(P＜0.05),与年龄、性别、病理类型无明显相关(P＞0.05).结论 甲状腺肿瘤患者外周血Th22细胞比例及血浆中IL-22水平显著升高,且与临床分期等病理特征明显相关,表明Th22细胞及IL-22参与甲状腺肿瘤的发生发展.     

CD4+CD25-T淋巴细胞转化为CD4-CD8-调节T细胞的体外实验研究

目的 探讨体外诱导和纯化CD4+ CD25-T淋巴细胞(effector T cell,Teff)转化为CD4-CD8-双阴性调节T细胞(double negative regulatory T cell,DN Treg)的最适条件.方法 采用免疫磁珠分选方法提取C57BL/6小鼠的CD4+ CD25-T淋巴细胞、DBA/2小鼠的成熟树突状细胞共培养,加入不同剂量的IL-2,通过流式细胞检测CD4-CD8-T细胞的转化比例并确定最适条件,免疫磁珠阴性选择分选提纯转化的CD3+ CI4-CD8-T细胞,流式细胞仪检测转化的CD4-CD8-调节T细胞对CD4+ CD25-效应T细胞增殖抑制情况.结果 CD4+ CD25-T淋巴细胞与DBA/2小鼠的树突状细胞共培养6d后检测CD4-CD8-调节T细胞的转化比率为6.21％ ±2.03％,实验组加入不同浓度IL-2的转化率:A组(25 ng/ml)为14.77％±2.15％,B组(50 ng/ml)为21.29％±2.68％,C组(75 ng/ml)为43.45％ ±4.45％,D组(100 ng/ml)为28.59％±3.05％,IL-2浓度在75 ng/ml时,转化获得率最高(C组与对照组、实验A、B、D组比较分别t=10.700,8.288,6.158,3.932,均P＜0.05);分离提纯CD4-CD8-双阴性调节细胞纯度达到98.10％,CD4-CD8-双阴性调节细胞与CFSE染色的CD4+ CD25-T淋巴细胞、小鼠树突状细胞共培养6d,实验组增殖指数为1.15明显低于对照组2.07.结论 小鼠CD4+ CW25-T淋巴细胞在体外,成熟树突状细胞刺激下可转化为CD4-CD8-双阴性调节T细胞,IL-2可显著提高其转化率.     

The relationship between CD4-CD8-T cells in the peripheral blood of patients with pancreatic carcinoma and IL-4,IFN-γ levels

Objective To investigate the relationship between CD4-CD8-T cells ratio and IL-4, IFN-γ levels in the peripheral blood of patients with pancreatic carcinoma. Methods Floweytometer was used to analyze the CD4-CD8-T cells ratio in the peripheral blood of patients with pancreatic carcinoma and the IL-4 ,IFN-γ levels were detected by ELISA. Results The ratio of CD4-CD8-T cell in CD3+ T cell from 25 pancreatic carcinoma patients was(4.2 ± 1.7) %, the ratio of CD4-CD8-T cell in CD3+T cell from 45 healthy person was(6.3 ± 2.6) %, there was significant deviation between the two groups(P ＜ 0. 01). The IL-4 level of 25 pancreatic carcinoma patients was (86.3 ± 23.3) fg/L,the IL-4 level of 45 healthy person was (56.2 ± 9.2) fg/L,there was significant deviation between the two groups(P ＜0.01). The IFN-γ level of 25 pancreatic carcinoma patients was (16.4±4.8) fg/L before operation, the IFN-γ level of 45 healthy person was (27.4±3.8) fg/L,there was significant deviation between the two groups(P ＜0.01). The ratio of CD4-CD8-T cell in pancreatic carcinoma patient after operation was higher than before operation. It could be found negative correlation between CD4-CD8-T cells ratio and IL-4 level in pancreatic carcinoma patient,it could also be found positive correlation between CD4-CD8-T cells ratio and IFN-γ level in pancreatic carcinoma patient. In pancreatic carcinoma patient,the CD4-CD8-T cells ratio and IL-4 level was significant associated with clinical stage (P ＜ 0.05), but no relationship with hisological differentiation (P＞0.05), it could be found no relationship between IFN-γ level and clinical stage, hisoiogieal differentiation (P ＞ 0.05). Conclusion The CD4±CD8±T cells ratio in the peripheral blood of patients is decreased,it may be participate in the carcinogenesis and development of pancreatic carcinoma by influence the IFN-γ levels.     

The relationship between CD4-CD8-T cells in the peripheral blood of patients with pancreatic carcinoma and IL-4,IFN-γ levels

Objective To investigate the relationship between CD4-CD8-T cells ratio and IL-4, IFN-γ levels in the peripheral blood of patients with pancreatic carcinoma. Methods Floweytometer was used to analyze the CD4-CD8-T cells ratio in the peripheral blood of patients with pancreatic carcinoma and the IL-4 ,IFN-γ levels were detected by ELISA. Results The ratio of CD4-CD8-T cell in CD3+ T cell from 25 pancreatic carcinoma patients was(4.2 ± 1.7) %, the ratio of CD4-CD8-T cell in CD3+T cell from 45 healthy person was(6.3 ± 2.6) %, there was significant deviation between the two groups(P ＜ 0. 01). The IL-4 level of 25 pancreatic carcinoma patients was (86.3 ± 23.3) fg/L,the IL-4 level of 45 healthy person was (56.2 ± 9.2) fg/L,there was significant deviation between the two groups(P ＜0.01). The IFN-γ level of 25 pancreatic carcinoma patients was (16.4±4.8) fg/L before operation, the IFN-γ level of 45 healthy person was (27.4±3.8) fg/L,there was significant deviation between the two groups(P ＜0.01). The ratio of CD4-CD8-T cell in pancreatic carcinoma patient after operation was higher than before operation. It could be found negative correlation between CD4-CD8-T cells ratio and IL-4 level in pancreatic carcinoma patient,it could also be found positive correlation between CD4-CD8-T cells ratio and IFN-γ level in pancreatic carcinoma patient. In pancreatic carcinoma patient,the CD4-CD8-T cells ratio and IL-4 level was significant associated with clinical stage (P ＜ 0.05), but no relationship with hisological differentiation (P＞0.05), it could be found no relationship between IFN-γ level and clinical stage, hisoiogieal differentiation (P ＞ 0.05). Conclusion The CD4±CD8±T cells ratio in the peripheral blood of patients is decreased,it may be participate in the carcinogenesis and development of pancreatic carcinoma by influence the IFN-γ levels.     

The relationship between CD4-CD8-T cells in the peripheral blood of patients with pancreatic carcinoma and IL-4,IFN-γ levels

Objective To investigate the relationship between CD4-CD8-T cells ratio and IL-4, IFN-γ levels in the peripheral blood of patients with pancreatic carcinoma. Methods Floweytometer was used to analyze the CD4-CD8-T cells ratio in the peripheral blood of patients with pancreatic carcinoma and the IL-4 ,IFN-γ levels were detected by ELISA. Results The ratio of CD4-CD8-T cell in CD3+ T cell from 25 pancreatic carcinoma patients was(4.2 ± 1.7) %, the ratio of CD4-CD8-T cell in CD3+T cell from 45 healthy person was(6.3 ± 2.6) %, there was significant deviation between the two groups(P ＜ 0. 01). The IL-4 level of 25 pancreatic carcinoma patients was (86.3 ± 23.3) fg/L,the IL-4 level of 45 healthy person was (56.2 ± 9.2) fg/L,there was significant deviation between the two groups(P ＜0.01). The IFN-γ level of 25 pancreatic carcinoma patients was (16.4±4.8) fg/L before operation, the IFN-γ level of 45 healthy person was (27.4±3.8) fg/L,there was significant deviation between the two groups(P ＜0.01). The ratio of CD4-CD8-T cell in pancreatic carcinoma patient after operation was higher than before operation. It could be found negative correlation between CD4-CD8-T cells ratio and IL-4 level in pancreatic carcinoma patient,it could also be found positive correlation between CD4-CD8-T cells ratio and IFN-γ level in pancreatic carcinoma patient. In pancreatic carcinoma patient,the CD4-CD8-T cells ratio and IL-4 level was significant associated with clinical stage (P ＜ 0.05), but no relationship with hisological differentiation (P＞0.05), it could be found no relationship between IFN-γ level and clinical stage, hisoiogieal differentiation (P ＞ 0.05). Conclusion The CD4±CD8±T cells ratio in the peripheral blood of patients is decreased,it may be participate in the carcinogenesis and development of pancreatic carcinoma by influence the IFN-γ levels.     

Immunosuppressive effect on T cell activation by interleukin-16- and interleukin-10-cDNA-double-transfected human squamous cell line

It is well known that induction of immunotolerance with allogeneic skin transplantation is generally difficult. This study attempted to find an immunosuppressive protocol for skin allograft rejection involving interleukin-16 (IL-16) and interleukin-10 (IL-10), because both are known to inhibit mixed lymphocyte reaction (MLR). The data indicated that IL-16 enhanced the immunosuppressive effect of IL-10. IL-16-cDNA- and IL-10-cDNA-double-transfected squamous cell carcinoma cell line were used as an in vitro model and they produced more than 20 ng/ml of IL-16 and 100 pg/ml of IL-10 in the supernatant, which significantly inhibited MLR and also the activation of allogeneic lymphocytes, which were stimulated directly by allogeneic double-cDNA-transfectant cells. Thus allogeneic skin graft producing IL-16 and IL-10 might have a local immunosuppressive action that could prolong graft survival.     

Osteoblasts derived from osteophytes produce interleukin-6, interleukin-8, and matrix metalloproteinase-13 in osteoarthritis

To clarify the significance of the osteophytes that appear during the progression of osteoarthritis (OA), we investigated the expression of inflammatory cytokines and proteases in osteoblasts from osteophytes. We also examined the influence of mechanical stress loading on osteoblasts on the expression of inflammatory cytokines and proteases. Osteoblasts were isolated from osteophytes in 19 patients diagnosed with knee OA and from subchondral bone in 4 patients diagnosed with femoral neck fracture. Messenger RNA expression and protein production of inflammatory cytokines and proteases were analyzed using real-time RT-PCR and ELISA, respectively. To examine the effects of mechanical loading, continuous hydrostatic pressure was applied to the osteoblasts. We determined the mRNA expression and protein production of IL-6, IL-8, and MMP-13, which are involved in the progression of OA, were increased in the osteophytes. Additionally, when OA pathological conditions were simulated by applying a nonphysiological mechanical stress load, the gene expression of IL-6 and IL-8 increased. Our results suggested that nonphysiological mechanical stress may induce the expression of biological factors in the osteophytes and is involved in OA progression. By controlling the expression of these genes in the osteophytes, the progression of cartilage degeneration in OA may be reduced, suggesting a new treatment strategy for OA.     

Dorsal root sensitivity to interleukin-1 beta, interleukin-6 and tumor necrosis factor in rats

The release of inflammatory cytokines caused by a disrupted disc may play a critical role in pain production at nerve endings, axons, and nerve cell bodies. Herniated disc tissue has been shown to release inflammatory cytokines such as interleukin-1 beta (IL-1beta), interleukin-6 (IL-6), tumor necrosis factor (TNF), and other algesic chemicals. This study was designed to characterize the effects of these proinflammatory cytokines on the somatosensory neural response at the dorsal root level in rats. It is hypothesized that their effects on nerve endings in disc and adjacent tissue contribute to low-back pain, and the effects on dorsal root axons and ganglia contribute to radiculopathy and sciatica. Surgically isolated sacral dorsal roots were investigated by electrophysiologic techniques. IL-1beta, IL-6, or TNF (100 ng, each) were applied onto the dorsal roots. Neural responses and mechanosensitivity of the receptive fields were evaluated over time. The results showed that 3 h after each cytokine application, the neural activity was statistically decreased. The mechanical sensitivity of the receptive fields increased at 90 min following IL-1beta or TNF application, and returned to normal more than 3 h after IL-1beta application. IL-1beta, IL-6, and TNF may be neurotoxic to dorsal root axons. Furthermore IL-1beta and TNF may sensitize the peripheral receptive fields. This study suggests that dorsal roots may be impaired by these proinflammatory cytokines.     

全髋关节置换术后血清白细胞介素-6、10与纤维蛋白原表达的特点分析

目的 探讨血清炎性因子白细胞介素(IL)-6、IL-10及纤维蛋白原(FBG)浓度在人工全髋关节置换术(THA)后的变化特点及意义.方法 选择初次行单侧THA的30例患者为观察组,其中男12例,女18例;年龄55～70岁(平均58.4岁),分别于术前1 d、术后1、3 d检测IL-6、IL-10及FBG浓度,并与对照组20例健康人群(男8例,女12例;年龄25～40岁,平均30.5岁)结果进行比较.结果 观察组患者THA后1 d血清中IL-6、IL-10及FBG浓度分别为(121.74±84.93)pg/mL、(14.92±11.80)pg/mL、(4833.2±314.3)mg/L,较对照组[(41.21±25.35)pg/mL、(8.14±7.25)pg/mL、(3097.2±771.3)mg/L]及术前1 d[(47.41±76.32)pg/mL、(8.30±10.69)pg/mL、(3497.2±677.2)mg/L]明显升高,差异均有统计学意义(P＜0.05＝.术后3 d血清中IL-6、IL-10及FBG浓度较术后1 d明显下降(P＜0.05＝,基本接近术前水平,与术前1 d比较差异均无统计学意义(P＞0.05).结论 炎症与血栓形成密切相关,THA后3 d是抗炎、抗凝治疗的关键.术后及时监测IL-6、IL-10及FBG浓度的变化,同时观察临床症状及生化指标,对于更好地掌握病情的变化及预防血栓形成具有一定临床意义.

Abstract：

Objective To investigate changes in the expressions of inflammatory cytokines (IL-6,IL-10) and fibrinogen (FBG) in blood serum in patients after total hip arthroplasty(THA) .Methods Thirty patients who underwent primary THA at one side were enrolled in the experimental group.They were 12 men and 18 women, aged from 55 to 70 years (mean, 58.4 years).Expressions of IL-6, IL-10 and FBG in blood serum at one day preoperation and 1, 3 days postoperation were detected respectively by ELISA.Their values were compared with the normal values of 20 healthy controls [8 men and 12 women, with a mean age of 30.5 years (from 25 to 40 years)].Results The serum IL-6, IL-10 and FBG in the experimental group one day after THA were respectively 121.74 ±84.93 pg/mL, 14.92 ± 11.80 pg/mL and 4833.2 ±314.3 mg/L, significantly higher than those one day preoperation (47.41 ±76.32 pg/mL, 8.30 ± 10.69 pg/mL and 3497.2 ± 677.2 mg/L) and those in the control group (41.21 ± 25.35 pg/mL, 8.14 ± 7.25 pg/mL and 3097.2 ±771.3 mg/L) ( P ＜ 0.05).The values 3 days postoperation significantly decreased compared to those one day postoperation ( P ＜ 0.05 ), similar to those one day preoperation.There was no significant difference between 3 days postoperation and one day preoperation ( P ＞ 0.05) .Conclusions Since inflammation is closely related to thrombosis, 1 to 3 days after THA may be the most crucial time for anti-inflammatory and anticoagulation management.To better observe treatment outcome and predict prognosis, it is essential to closely monitor IL-6, IL-10 and FBG, as well as life signs at the same time after operation.     

Lymphocyte subpopulations,interleukin-2 and interleukin-2 receptor expression in childhood nephrotic syndrome

Abnormal T lymphocyte function and reduced interleukin-2 (IL-2) production have been implicated in the pathogenesis of the nephrotic syndrome (NS). We investigated: (1) lymphocyte subpopulations and expression of IL-2 receptor (IL-2R) on T cells using two-colour flow cytometry, (2) serum IL-2 and (3) the soluble component of IL-2R (sIL-2R) in serum, using enzyme-linked immunosorbent assay, in 38 children with NS. All children, except those in remission, had marked proteinuria. They were divided into groups according to renal pathology: (1) steroid-sensitive NS (SSNS) not receiving prednisolone therapy, (2) SSNS on prednisolone, (3) focal segmental glomerulosclerosis (FSGS), (4) SSNS in remission and not receiving prednisolone therapy, (5) congenital NS (CNS). Results were compared with 26 age-matched controls. Total T lymphocytes (CD3) were reduced in groups 1 and 2; CD4 count was reduced in groups 1–4; CD8 count increased in groups 2 and 3; CD8 and CD19 (B lymphocytes) were significantly reduced in group 5. Increased IL-2R expression (CD25) on CD4 lymphocytes was noted in groups 1, 2 and 3 implying activation of these cells. In patients with SSNS, increased serum sIL-2R was recorded during relapse (1,273±497 U/l vs. 913±401 U/l in remission,P<0.005) but free serum IL-2 was not detectable at any stage. The specific alterations in lymphocyte subpopulations in SSNS and FSGS would imply an involvement of the immune system distinct from that in CNS.     

T-cell subsets,interleukin-2 receptor expression and production of interleukin-2 in minimal change nephrotic syndrome

This study was designed to investigate T-lymphocyte subsets interleukin-2 receptor (IL-2R) expression and IL-2 production in minimal change nephrotic syndrome (MCNS). Peripheral blood T-lymphocytes and IL-2R expression were analysed using fluorescein isothiocyanatelabelled CD3, CD4, CD8 and CD25 monoclonal antibodies with flow cytometry. IL-2 production was determined by enzyme immunoassay. Ten children with MCNS in relapse and in remission were evaluated. Thirteen healthy children served as controls. The patients in relapse demonstrated a moderate decrease in the total absolute lymphocyte counts and CD8(+) T-lymphocytes compared with controls (P<0.05) and had a greatly increased IL-2R expression in frashly isolated, unstimulated peripheral lymphocytes compared with patients in remission and controls. While this was not statistically significant, IL-2R expression on cultured lymphocytes stimulated with phytohaemagglutinin was significantly elevated in relapse compared with those in remission and controls (P<0.05). IL-2 production did not correlate well with IL-2R expression and there was no significant difference between the groups. Our results suggest that T-cell subset changes and high IL-2R expression on peripheral lymphocytes may indicate the presence of stimulated T-cell populations in MCNS which could contribute to the immunopathogenesis.     

白细胞介素-17和白细胞介素-6在肝内胆管细胞癌的表达及其临床意义

目的 探讨肝内胆管细胞癌(ICC)白细胞介素-17(IL-17)和白细胞介素-6(IL-6)表达的临床意义.方法 采用免疫组织化学Envision二步染色法检测69例ICC肿瘤组织和63例癌旁组织IL-17和IL-6的表达,分析两者的关系及其与临床病理特征和预后的关系.结果 肿瘤和癌旁组织中IL-17+细胞密度中位值分别为36.0/HP(每高倍视野)和8.0/HP;肿瘤组织IL-17低表达组和高表达组IL-6阳性表达率分别为46.9％和75.0％;IL-17和IL-6的表达呈正相关(r=0.256,P＜0.05);肿瘤组织IL-17低表达组术后生存时间较长(P＜0.01),Cox多因素分析提示肿瘤组织IL-17表达水平( HR=2.462,P＜0.05)是ICC患者术后的独立预后因素.结论 ICC肿瘤组织IL-17表达与IL-6表达呈正相关,与ICC患者术后生存呈负相关,可作为预测患者预后的指标.     

高迁移率族蛋白B1对小鼠调节性T细胞与CD4+CD25-T细胞相互作用的影响

目的 观察高迁移率族蛋白B1(HMGB1)对调节性T细胞(Treg)与CD4+CD25-T细胞相互作用的影响,并初步探讨其影响Treg抑制功能的机制.方法 免疫磁珠法分离正常BALB/c小鼠脾脏CD4+CD25+Treg及CD4+CD25-T细胞.采用固相包被抗CD3/可溶性抗CD28进行辅助活化,以不同时间及浓度HMGB1刺激Treg,ELISA法分析HMGB1刺激对Treg分泌IL-10的影响.将HMGB1(1000μg/L)刺激后的Treg与CD4+CD25-T细胞共培养,MTT法观察其对Treg抑制CD4+CD25-T细胞反应的作用,并分析HMGB1刺激后的Treg对CD4+CD25-T细胞IL-2生成及细胞功能极化的影响.结果 经抗CD3/CD28辅助活化的CD4+CD25+Treg在HMGB1作用下IL-2生成无显著差异(P＞0.05),但随时间延长及剂量增加,IL-10生成明显减少(P＜0.05).经HMGB1刺激的Treg对CD4+CD25-T细胞增殖的抑制反应减弱,同时诱导CD4+CD25-T细胞IL-2产生及细胞功能极化的能力均下降(P＜0.05).结论 HMGB1可通过诱导Treg抑制功能的下调,从而影响CD4+CD25-T细胞功能,进而调节炎症反应过程.     

Expansion of CD3+CD56+ lymphocytes correlates with induction of cytotoxicity by interleukin-2 gene transfer in human breast tumor cultures

Background: Mice immunized with murine mammary carcinoma cells genetically engineered to secrete interleukin-2 (IL-2) are rendered resistant to subsequent challenge with unmodified tumor cells, and in the case of mice bearing established tumors, the rate of development of pulmonary metastases is reduced. Despite these encouraging animal results, little is known about the induction of antitumor immunity by IL-2 gene transfer in human breast cancer. Methods: Adenovirally mediated IL-2 gene transfer was performed in 12 tumor fragment cultures established from seven primary breast cancers. Autologous tumor infiltrating lymphocytes (TILs) or peripheral blood mononuclear cells (PBMCs) were cocultured with transduced tumor fragments, and changes in phenotype and cytotoxicity were measured. Results: IL-2 was never detectable in the untransduced cultures, but it peaked at 5.0—1,324.8 ng/ml in the transduced cultures. Lymphocyte counts declined in all untransduced cultures, but they increased two- to sevenfold in four transduced cultures. CD4:CD8 ratios decreased from a mean of 2.11 at baseline to 1.27 after stimulation in coculture (p=0.03). Expansion of lymphocytes expressing the natural killer cell phenotype (CD3⁻CD56⁺) occurred in only one culture, but the CD3⁺CD56⁺ population increased in four of six cultures. Lymphocytes from four of 10 cocultures generated significant cytotoxicity against allogeneic breast cancer cells. Induction of cytotoxicity correlated with expansion of the CD3⁺CD56⁺ phenotype (R²=0.805, p=0.02). Conclusions: IL-2 gene expression by human breast cancer causes expansion of CD3⁺CD56⁺ cytotoxic lymphocytes. This phenotype is consistent with that of a non-major histocompatibility complex (MHC)-restricted cytokine induced killer cell population previously described. Opinions, interpretations, conclusions and recommendations are those of the author and are not necessarily endorsed by the U.S. Army. Presented at the 49th Annual Cancer Symposium of The Society of Surgical Oncology, Atlanta, Georgia, March 21–24, 1996.     

白细胞介素-6和白细胞介素-10在不同细菌性血流感染小鼠模型中的表达及意义

目的:探讨白细胞介素(IL)-6和IL-10在不同病原菌所致细菌性血流感染中的表达及其意义.方法:建立金黄色葡萄球菌、粪肠球菌、大肠杆菌和肺炎克雷伯菌ICR小鼠血流感染模型,以注射相应剂量磷酸盐缓冲液的小鼠作为对照组,采用Luminex液相悬浮芯片系统检测各实验组和PBS对照组感染后0.5、1、3、6、12、24和48h时IL-6和IL-10的含量.结果:细菌感染小鼠后0.5 h时,各实验组IL-6的含量与对照组相比均明显升高(P＜0.05).各实验组IL-10的水平与对照组相比,感染后0.5h均明显升高(P＜0.05).大肠杆菌组、肺炎克雷伯菌组IL-10的含量在感染后1h达最大值.金黄色葡萄球菌组感染后3h达到最大值,粪肠球菌组在感染后6h达到最大值,且大肠杆菌组和肺炎克雷伯菌组IL-6和IL-10含量升高的幅度明显高于金黄色葡萄球菌组和粪肠球菌组(P＜0.05).对照组小鼠血清中IL-6和IL-10的含量在此过程中无明显变化.结论:IL-6和IL-10的含量在4种细菌性血流感染模型早期均明显升高.大肠杆菌组、肺炎克雷伯菌组IL-6和IL-10的含量升高幅度明显高于金黄色葡萄球菌组和粪肠球菌组.联合运用IL-6和IL-10有可能用于区分革兰阳性或革兰阴性细菌感染.