白细胞介素2激活骨髓抗肿瘤机理的实验研究

目的　探讨白细胞介素 2 (IL 2 )激活的骨髓 (ABM )抗肿瘤作用的免疫学机理。方法　取恶性血液病缓解期患者骨髓 ,分离单个核细胞 (MNC) ,与IL 2共同孵育后 ,测定CD3-CD86 、CD2 8 、CD8 CD2 8 、CD3-CD16 /CD5 6 的细胞比例及上清液中肿瘤坏死因子α(TNF α) ,干扰素γ(IFN γ)的含量。结果　 (1)培养 72小时后 ,实验组CD3-CD86 细胞、CD2 8 细胞、NK细胞比例及上清中TNF α和IFN γ的含量均比对照组增高 (P <0 0 5 )。 (2 )培养 1周后 ,实验组CD3-CD86 比对照组显著增高 (P <0 0 1) ,CD2 8 细胞、NK细胞比例、CD8 CD2 8 均增高 (P <0 0 5 )。结论　 (1)ABM中CD3-细胞表面共刺激分子CD86表达明显增加 ,与ABM中特异性细胞毒性T淋巴细胞(CTLs)的生成有关。 (2 )ABM中活化B细胞、NK细胞、CTLs、TNF α和IFN γ也直接或间接地参与了ABM的抗肿瘤作用     

rhIL-10对银屑病动物模型的治疗作用及免疫功能的影响

目的　研究重组人白细胞介素 10 (rhIL 10 )对银屑病动物模型的治疗作用以及免疫功能的影响。方法　用小鼠雌激素期阴道上皮模型、鼠尾鳞片表皮模型观察重组人白细胞介素 10的抗银屑病作用 ,并检测ConA诱导T淋巴细胞增殖、LPS诱导B淋巴细胞增殖、NK细胞活性及细胞因子(IFN γ、IL 2、IL 1、TNF α)水平。结果　rhIL 10 (5 ,2 0 ,80μg·kg-1·d-1,sc)对小鼠阴道上皮细胞有丝分裂有抑制作用 ,促进小鼠尾鳞片颗粒层的形成。对ConA诱导T淋巴细胞增殖反应及IL 2、IFN γ产生有抑制作用 ;对LPS诱导小鼠腹腔巨噬细胞产生IL 1、TNF α具有抑制作用 ;对NK细胞活性有抑制作用 ;对LPS诱导B淋巴细胞增殖有促进作用。结论　rhIL 10可能是通过抑制表皮细胞增殖与促进其分化 ,并参与抗炎与免疫调节过程而发挥治疗银屑病动物的作用     

猴头多糖抗肿瘤及对免疫功能的影响

目的　研究猴头多糖 ( HEPS)对小鼠 S1 80 肉瘤及免疫功能的影响。方法　 H EPS按 10 0、2 0 0、40 0m g/ kg体重连续灌胃 15 d,测定荷瘤小鼠瘤重 ,通过检测小鼠抗体生成细胞、迟发性变态反应、NK细胞活性 ,荷瘤小鼠免疫器官分别检测 HEPS对体液免疫、细胞免疫、非特异免疫及异常免疫的调节作用。结果　 HEPS可显著抑制 S1 80 肉瘤的生长 ,提高荷瘤小鼠胸腺和脾重 ,增强正常小鼠抗体形成细胞溶解绵羊红细胞能力、迟发型变态反应能力、NK细胞活性。结论　 HEPS具有抗肿瘤及免疫调节作用     

重组人胸腺素α原体外对IFN-γ、IFN-α及TNF-α的影响

目的　在体外实验中考察克隆并表达的人胸腺素α原 (prothymosinα ,ProTα)对几种重要细胞因子IFN γ、IFN α和TNF α分泌的影响。方法　在体外分离脾淋巴细胞、脾巨噬细胞及腹腔巨噬细胞 ,用ELISA法检测ProTα对IFN γ、IFN α和TNF α分泌的影响。结果　 10 - 7mol·L- 1 ProTα明显促进脾细胞分泌IFN γ(P <0 .0 5 ) ,该浓度的ProTα也明显刺激小鼠脾巨噬细胞分泌IFN α(P <0 .0 1) ;在小鼠腹腔巨噬细胞中 ,ProTα能明显刺激IFN α和TNF α的分泌 (P <0 .0 1)。结论　在体外实验中 ,ProTα对细胞因子IFN γ、IFN α和TNF α的分泌均有促进作用 ,这为ProTα进一步体内研究奠定了基础     

卡介菌多糖核酸对肺结核病人外周血细胞因子mRNA表达的调节

目的 :观察活动性肺结核病人外周血单个核细胞 (PBMC)细胞因子信息核糖核酸 (mRNA)表达特点以及卡介菌多糖核酸的调节作用。方法 :活动性肺结核病人和健康志愿者各 15例 ,分离单个核细胞。分 3组进行培养 ,A组为不加刺激物的对照组 ,B组加入结核菌素纯蛋白衍生物 (PPD) ,C组加入卡介菌多糖核酸 (BCG PSN )。检测PBMCγ 干扰素 (IFN γ) ,肿瘤坏死因子 (TNF α)和白细胞介素10 (IL 10 )mRNA表达。结果 :活动性肺结核病人PBMC在A组IFN γmRNA的表达明显低于健康者 ,P <0 .0 1,TNF α与IL 10mRNA的表达与健康人无差异。在B组IFN γmRNA和TNF αmRNA表达与健康人无差异 ;IL 10mRNA的表达增强 ,与健康人比较 ,P <0 .0 1。在C组 ,IFN γmRNA的表达增强 ,仍低于健康人 ,P <0 .0 5 ;TNF αmRNA的表达有增加 ,与健康人比较 ,P >0 .0 5 ,IL 10mR NA的表达无改变。结论 :活动性肺结核病人保护性细胞因子IFN γ减低 ;BCG PSN刺激可提高免疫细胞IFN γmRNA ,但不增高IL 10mRNA的生成     

活动期类风湿关节炎患者血清可溶性细胞间粘附分子-1 E-选择素变化及意义

目的　测定活动期类风湿关节炎 (RA)患者血清中可溶性细胞间粘附分子 (sICAM) 1、可溶性E 选择素 (sE 选择素 )、白细胞介素 (IL) 1、肿瘤坏死因子 (TNF)、干扰素 (IFN) γ水平 ,探讨sICAM 1、sE 选择素与IL 1、TNF、IFN γ及病情的关系。方法　用酶联免疫分析法 (ELISA)检测 3 0例活动期RA患者与 3 0名正常对照者sICAM 1、sE 选择素、IL 1、TNF、INF γ水平。结果　RA患者血清sICAM 1、sE 选择素、IL 1、TNF、IFN γ水平明显高于正常对照组 (P <0 0 0 1) ,sICAM 1与sE 选择素、IL 1、IFN γ水平及类风湿因子 (RF)正相关 ,sE 选择素与IL 1正相关。结论　RA患者血清sICAM 1、sE 选择素、IL 1、TNF、IFN γ水平显著升高 ,ICAM 1及sE 选择素参与RA发病过程 ,sICAM 1可作为判断病情严重性的指标     

中药抑瘤胶囊对荷瘤小鼠免疫系统的作用

目的观察中药抑瘤胶囊对荷瘤小鼠免疫系统的影响。方法检测胸腺细胞自然增殖能力，脾细胞诱导的白细胞介素-2（IL-2）、干扰素（IFN）的产生及活性，自然杀伤细胞（NK）杀伤活性等四项指标，观察中药抑瘤胶囊对荷瘤小鼠免疫系统的作用。结果中药抑瘤胶囊可促进小鼠胸腺细胞自然增殖能力，受试药三个剂量组与对照组比较，P〈0．05；对小鼠脾细胞诱导产生的IFN活性随剂量增高而增强，各剂量组与对照组比较，P〈0．05；该药对小鼠NK细胞杀伤活性的作用也与剂量呈正相关，但对小鼠脾细胞诱导产生的IL-2活性随剂量增高而呈下降趋势，与对照组比较，只有低剂量组IL-2活性增强作用明显，P〈0．05。结论中药抑瘤胶囊对荷瘤小鼠免疫系统功能有明显增强作用。     

绞股蓝总皂甙对Lewis肺癌荷瘤小鼠肿瘤生长及免疫功能的影响

目的 :观察绞股蓝总皂甙 (gypenosides,GPs)对Lewis肺癌荷瘤小鼠肿瘤生长、脾淋巴细胞数及NK活性的影响。方法 :整体动物的抑瘤试验、脾淋巴细胞计数及NK活性测定。结果 :GPs对荷瘤小鼠Lewis肺癌细胞具有明显的抑制作用 ,在剂量 1 0、2 0、4 0mg/kg腹腔注射 (ip)给药条件下 ,其抑瘤率分别为 (3 0 7± 1 2 ) % ,(51 5± 2 5) %和 (50 3± 1 5) % (P <0 0 1 )。同时 ,GPsip给药后荷瘤小鼠脾淋巴细胞总数明显增加 ,外周血淋巴细胞NK活性明显升高 ,脾淋巴细胞NK活性也明显升高。结论 :GPs的抗肿瘤作用与其对荷瘤小鼠免疫功能的增强作用密切相关。     

喇咕来源水溶性壳聚糖抗肿瘤作用及其免疫调节活性研究

目的从喇咕中提取制备水溶性壳聚糖 ,并研究其抗肿瘤作用及免疫活性。方法按 2 0 0mg (kg·d)剂量灌胃给药 ,观察其对荷瘤小鼠的抑瘤作用和免疫学指标。结果水溶性壳聚糖对S180 和艾氏腹水癌的抑瘤率分别为 58.6%和 54.85% ,与环磷酰胺伍用 ,能增强荷瘤小鼠抑瘤率 ;并能拮抗环磷酰胺所致的副作用。结论喇咕来源水溶性壳聚糖对荷瘤小鼠有显著的抗肿瘤作用 ,并能调节机体免疫功能 ,降低化疗药物的毒副作用。     

慢性阻塞性肺病血清IL-6、TNF-α、I FN-γ的作用及意义

目的 :探讨血清白细胞介素6(IL 6)、肿瘤坏死因子α(TNF α)和干扰素γ(IFN γ)在慢性阻塞性肺病(COPD)气道炎症发病中的作用。方法 :采用双抗体夹心ABC ELISA法对COPD(COPD组 )、肺气肿、COPD合并肺心病 (肺心病组 )及正常对照组血清IL 6、TNF α、IFN γ进行测定。结果 :肺心病组和COPD组血清IL 6、TNF α水平均明显高于对照组 (P<0 05) ,血清IFN γ水平明显低于对照组 (P<0 05)。且2组患者血清IL 6与TNF α呈正相关 (P<0 05) ;肺心病组血清IL 6、TNF α水平均高于COPD组 ,但差别无统计学意义。2组患者吸烟组IL 6、TNF α水平均显著高于非吸烟组 (P<0 05)。吸烟年限、每日吸烟量与IFN γ呈负相关 (P<0 05) ,与TNF α呈正相关 (P<0 05)。结论 :IL 6、TNF α、IFN γ参与COPD患者气道炎症反应 ,吸烟可加快COPD气道炎症的进展。     

干扰素-γ免疫脂质体靶向治疗大鼠小细胞肺癌的实验研究

目的观察干扰素-γ免疫脂质体应用于大鼠小细胞肺癌靶向治疗的疗效。方法将干扰素-γ免疫脂质体应用于大鼠小细胞肺癌的治疗,观察治疗效果。结果比较两组大鼠治疗后第3,7,10天肿瘤生长速度,观察组明显低于对照组,差异显著(P<0.05);观察组大鼠平均生存时间、肺转移结节数与对照组相比均有明显差异(P<0.05);观察组肿瘤组织微血管密度与对照组相比有明显差异(P<0.05);观察组脾脏CTL细胞毒性、NK细胞毒性及TNF细胞毒性与对照组相比均有明显差异(P<0.05)。结论干扰素-γ免疫脂质体应用于小细胞肺癌大鼠的治疗,对肿瘤抑制作用强,治疗效果好,对组织损伤小,能延长大鼠生存时间。     

The source of Mycobacterium tuberculosis-specific IFN-γ production in peripheral blood mononuclear cells of TB patients

Mycobacterium tuberculosis (Mtb)-specific IFN-γ secretion plays important roles in anti-tuberculosis (TB) immunity. Mtb-specific IFN-γ response can be induced in HIV/TB co-infected patients with a low CD4 lymphocyte count; this suggests that the source of Mtb-specific IFN-γ production is not limited in CD4⁺ T lymphocytes. Currently, the major sources of Mtb-specific IFN-γ production and the function and phenotype of Mtb-specific IFN-γ-producing cells still remain unclear. Thirty-nine participants (24 active TB patients, 10 HIV/TB co-infected patients, and 5 healthy volunteers) were recruited according to conventional tests and Mtb-specific IFN-γ ELISPOT assay. Multicolor flow cytometry was used to investigate the production of intracellular IFN-γ in peripheral blood mononuclear cells (PBMCs) after Mtb-specific antigen stimulation. Our results showed that CD4⁺, CD8⁺ T cells and NK cells are all major sources of Mtb-specific IFN-γ production in PBMCs of TB patients. Moreover, CD8⁺ T cells are the highest number of Mtb-specific IFN-γ-producing cells in HIV/TB co-infected patients. Although the activity of NK cells is significantly reduced in TB patients when compared with healthy controls, Mtb-specific antigen stimulation induces a significant increase in NK cell activity. We also showed that CD45RO is the characteristic marker of Mtb-specific IFN-γ-producing T cells but not that of Mtb-specific IFN-γ-producing NK cells in peripheral blood. High expression of CD11a may be the characteristic feature of Mtb-specific IFN-γ-producing NK cells. This study put forward a new insight on the source of antigen-specific IFN-γ-production in PBMCs of TB patients.     

Promotion of interferon-gamma production by natural killer cells via suppression of murine peritoneal macrophage prostaglandin E2 production using intravenous anesthetic propofol

Propofol is an intravenous anesthetic, widely used for general anesthesia during surgery, which inevitably involves tissue trauma with inflammation. At sites of inflammation, prostanoids, especially prostaglandin E₂ (PGE₂), are abundant. This study addresses the effect of propofol on macrophage PGE₂ production. Using thioglycollate-elicited murine peritoneal macrophages, propofol (7.5–30 μM) suppressed lipopolysaccharide-induced PGE₂ production. The suppression was via the direct inhibition of cyclooxygenase (COX) enzyme activity and due neither to the downregulation of COX expression nor the inhibition of arachidonic acid release from plasma membranes. In macrophage:natural killer (NK) cell co-culture, propofol dramatically increased interferon-gamma (IFN-γ) production, and the actions of propofol were mimicked by a selective COX-2 inhibitor, NS-398, as well as the selective EP4 receptor antagonist L-161,982, suggesting a role of PGE₂ suppression in the upregulation of IFN-γ production. Furthermore, in purified NK cell culture, PGE₂ directly suppressed the production of IFN-γ by activated NK cells, which was reversed by selective inhibition of EP4 activity. Taken together, our results show that, in macrophage:NK cell co-culture, propofol, through the suppression of macrophage PGE₂ production, upregulates NK cell IFN-γ production by alleviating EP4 receptor-mediated suppression of IFN-γ production. Propofol may potentially exert considerable influence on inflammation and immunity by suppressing PGE₂ synthesis.     

Synergistic cytotoxicity of ex vivo expanded natural killer cells in combination with monoclonal antibody drugs against cancer cells

The adoptive transfer of highly cytotoxic natural killer (NK) cells is an emerging tool for cancer immunotherapy. Antibody-dependent cellular cytotoxicity (ADCC) has recently been identified as one of the critical factors for the clinical efficacy of anticancer antibodies, in which NK cells are the major effectors of ADCC. NK cells were expanded from PBMC by a feeder-cell-free expansion method. NK cell expansion efficiency was evaluated within a period of 21 days. The kinetics of NK cell expansion and the expression of activating and inhibitory receptors on NK cells were monitored. NK cells producing IFN-γ and TNF-α were detected by intracellular cytokine staining. The cytotoxicity of expanded NK cells against various cancer cells was compared with that of freshly isolated NK cells. The ADCC functions of expanded NK cells in combination with rituximab against CD20 + lymphoma cell lines were evaluated. Our method efficiently expanded NK cells ex vivo, which showed a much higher activity to induce the expression of activating receptors and to produce IFN-γ and TNF-α as well as cytotoxicity against various cancer cell lines including CD133 + primary cancer cells than freshly isolated NK cells. We observed a synergistic cytotoxicity of our expanded NK cells against CD20 + B lymphoma cell lines as well as higher IFN-γ and TNF-α production when combined with rituximab. Our results suggest that the adoptive transfer of a large number of ex vivo expanded NK cells, particularly in combination with monoclonal antibody drugs, is a useful tool for cancer immunotherapy.     

罗红霉素通过诱导型一氧化氮合酶/一氧化氮途径抑制哮喘大鼠气道炎症

目的探讨罗红霉素对哮喘大鼠支气管诱导型一氧化氮合酶(iNOS)及一氧化氮(NO)的影响。方法24只成年哮喘大鼠随机分成对照组、哮喘组以及罗红霉素组。对支气管肺泡灌洗液(BALF)细胞总数及嗜酸性粒细胞计数,免疫组织化学检测大鼠支气管上皮细胞iNOS蛋白表达,RT-PCR检测肺组织iNOS mRNA表达,分光光度计检测肺组织iNOS活性及NO含量。双抗体夹心法检测肺组织白细胞介素-4(IL-4)及干扰素-γ(IFN-γ)。结果哮喘组大鼠BALF细胞总数及嗜酸性粒细胞分类分别为(7.28±1.65)×108.L-1、(7.73±1.54)%,均高于对照组(3.76±0.97)×108.L-1、(1.27±0.60)%;罗红霉素组BALF细胞总数及嗜酸性粒细胞分类分别为(5.68±0.95)×108.L-1、(5.54±1.53)%,明显低于哮喘组,差异有统计学意义。哮喘组肺组织IL-4浓度、iNOS活性及NO含量高于对照组,罗红霉素组肺组织IL-4浓度、iNOS活性及NO含量低于哮喘组。哮喘组肺组织IFN-γ浓度低于对照组,罗红霉素组肺组织IFN-γ浓度高于哮喘组。哮喘大鼠支气管上皮细胞iNOS蛋白及肺组织iN-OSmRNA表达分布吸光度值分别为(0.25±0.06)、(0.52±0.14),较对照组[(0.14±0.05),(0.33±0.05)]明显增强;但罗红霉素组iNOS蛋白及mRNA表达为(0.15±0.03)、(0.35±0.07),均明显较哮喘组减弱。结论罗红霉素通过干预哮喘大鼠气道IL-4、IFN-γ以及iNOS/NO体系,抑制哮喘气道炎症反应。     

Immunomodulatory effect of diethylcarbamazine citrate plus filarial excretory–secretory product on rat hepatocarcinogenesis

Diethylcarbamazine citrate (DEC) had a significance in anti-filarial chemotherapy, while excretory–secretory product (ES) is released from adult filarial females. The target of the current study was to examine the immunomodulatory effect of DEC, Setaria equina ES or a combination of them on rat hepatocellular carcinoma (HCC) induced by diethylnitrosamine (DEN). In vitro effect of combined DEC and ES or ES alone on lipopolysaccharide (LPS)-stimulated rat peripheral blood mononuclear cells (PBMCs) was tested through IFN-γ assay in culture supernatants. In addition, single or repeated doses of DEC, ES or DEC + ES have been applied in white albino rats to test the effect on HCC. Levels of IFN-γ and anti-ES IgG antibodies in rat serum were assayed using ELISA. Hemolytic complement activity (CH₅₀) was determined in serum while the concentration of nitric oxide (NO) was assayed in liver tissue. The infiltration of NK cells as well as the expression of MHC Iproliferating cell nuclear antigen (PCNA), inducible NO synthase (iNOS), Bcl2 and p53 were determined using immunohistochemistry. There was a dose-dependent increase in IFN-γ after in vitro exposure to DEC + ES. Repeated ES doses increased NO concentration (p < 0.05) and expression of iNOS but reduced CH₅₀ (p < 0.001), while repeated DEC + ES doses could increase anti-ES IgG (p < 0.01), IFN-γ level (p < 0.05) and NK cell infiltration. The same treatments could also reduce the expression of MHC I expression, PCNA, Bcl2 and p53. This study has shown immunomodulatory and protective effects of DEC + ES repeated doses on rat HCC.     

Increased production of nitric oxide by neutrophils from patients with chronic granulomatous disease on interferon-gamma treatment

Chronic granulomatous disease (CGD) is a rare immunodeficiency disorder in which phagocytic leukocytes fail to generate superoxide (O(2)(-)) and antimicrobial oxidants. The therapeutic validity of interferon-gamma (IFN-γ) has been well established in CGD patients but its underlying mechanisms remain poorly understood. One probable mechanism has been suggested to be modulation of nitric oxide (NO) release from phagocytic cells. Herein, we investigated NO production from neutrophil cells in CGD patients on treatment with IFN-γ in vivo and in vitro. We measured NO levels in sera from 19 CGD patients (group I: 7 patients treated with TMP-SMX, group II: 12 patients treated with TMP-SMX and IFN-γ simultaneously) and healthy control individuals (8 cases). We also measured NO production from neutrophils in both patients groups as well as in control group after adding 100 U IFN-γ in vitro. Our results showed that there was a significant difference between the groups in the NO levels of serum; patients who received IFN-γ had significantly higher amount of NO than the other groups. Besides, NO levels increased significantly after adding 100 U IFN-γ in vitro in three studied groups, considerably in the patients on treatment with IFN-γ. As a brief conclusion, the effect of IFN-γ in increasing NO production is obvious. This could be an explanation for the therapeutic effect of IFN-γ in patients with CGD as NO acts as a bactericidal agent and plays a role in host defense mechanism instead of O(2)(-).     

A new reliable bioassay for determining the biological activity of human interleukin-12 by using human NK cell line NKG cells

A specific and accurate bioassay for determining the biological activity of human interleukin-12 (hIL-12), an important typical Th1 cytokine in both innate and adaptive immunity, is extensively required for biomedical and clinical study. In this paper, we used a new established NK cell line NKG cells as the responder to hIL-12 stimulation by detecting their IFN-γ production. It was found that NKG cells produced high level of IFN-γ when simulated by hIL-12, and the dose-response curve became the best Sigmoid curve (R(2)=0.9977, p<0.0001) when stimulated for 24h. The intra-assay CVs (<10%) and inter-assay CVs (<5%) demonstrated that the bioassay was precise and reproducible. Furthermore, no obvious cross-effects of other cytokines such as IL-2 and IL-18 was observed on the bioassay. Addition of the neutralizing anti-hIL-12 antibody to the bioassay significantly inhibited the IFN-γ production in a dose dependent manner, indicating that the bioactivity was actually mediated by hIL-12. The bioassay by using NKG cells was also suitable for determining the biological activity of recombinant hIL-12 in the form of purified product or culture supernatant by CHO-hIL-12 cell line. In conclusion, a reliable hIL-12 bioassay for determining its biological activity was established by using NKG cells as a responder and measuring their production of IFN-γ.     

糖尿病视网膜病变患者VEGF和IFN-γ及NO检测的临床价值

目的观察细胞生长因子与一氧化氮在2型糖尿病(DM)视网膜病变(DR)发病中的作用。方法采用酶联免疫吸附法和分光光度法检测74例DM患者血管内皮生长因子(VEGF)、γ-干扰素(IFN-γ)和一氧化氮(ON)的血浆含量,同时以30例健康人作为对照。结果①DM无眼底病变组(NDR)与视网膜病变组(DR)血浆VEGF、IFN-γ和NO含量均较对照组明显增高(P<0.05,P<0.05,P<0.01);②DR组VEGF含量明显高于NDR组(P<0.05),而IFN-γ和NO含量明显低于NDR组(P<0.01,P<0.05);③NDR组VEGF与NO含量呈直线正相关(r=0.30,P<0.05),其他各组均无相关性。结论VEGF与INF-γ过度表达在DM视网膜病变中起重要作用,NO代谢紊乱是导致DR发生中进展性微血管病变的重要因素。