Role of Low Dosage Arsenic Trioxide on Pulmonary Dendritic Cells in Asthmatic Mice

Objective: To investigate the distribution and recruitment of pulm onary dendritic cells (DCs) and the influence of low dosage arsenic trioxide (As 2O 3) on them in the airway of asthmatic mice. Methods: Thirty BALB/c mice were randomly divided into 3 groups: the control group, the asthmat ic group and the As 2O 3 treated group. The mice asthmatic model was induced v ia sensitizing with peritoneal injection of ovalbumin (OVA) for two times and th en provocated with aerosol inhalation of OVA for a week. The treated group was p eritoneally injected with 0.2 ml solution of As 2O 3 (4mg/kg) 0.5h after each provocation. The immunohistochemistry and computerised image analysis were appli ed to detect quantitatively the DCs in the lung and airway of mice. Resul ts: All intraepithelial nonlymphoid dendritic cells 145 (NLDC 145) throughout the respiratory tree in the mice of the control group formed a netwo rk with the density of DCs varying from (575±54) cells/mm 2 epithelial surface in the large airway, to (68±12) cells/mm 2 epithelial surface in the small ai rway. The distribution of airway NLDC 145 + in the asthmatic group was simil ar to that in the control group, but its density was significantly upregulated ( P <0.01). The distribution of airway NLDC 145 in the treated group was sim ilar to that in the asthmatic group, only its density was significantly downregu lated ( P <0.01). Conclusion: There is an integral network of N LDC 145 + throughout the respiratory tree. To downregulate the density but n ot change the distribution of pulmonary DCs could be an important therapeutic me chanism of low dosage As 2O 3 in treating asthma.

Hepatocellular carcinoma-specific immunotherapy with synthesized α1,3-galactosyl epitope-ulsed dendritic cells and cytokine-induced killer cells

AIM： To evaluate the safety and clinical efficacy of a new immunotherapy using both α-Gal epitope-pulsed dendritic cells （DCs） and cytokine-induced killer cells. METHODS： Freshly collected hepatocellular carcinoma （HCC） tumor tissues were incubated with a mixture of neuraminidase and recombinant αl,3-galactosyltrans- ferase （αI,3GT） to synthesize α-Gal epitopes on car- bohydrate chains of the glycoproteins of tumor mem- branes. The subsequent incubation of the processed membranes in the presence of human natural anti-Gal IgG resulted in the effective phagocytosis to the tumor membrane by DCs. Eighteen patients aged 38-78 years with stage 111 primary HCC were randomly chosen for the study; 9 patients served as controls, and 9 patients were enrolled in the study group.RESULTS： The evaluation demonstrated that the pro- cedure was safe; no serious side effects or autoimmune diseases were observed. The therapy significantly pro- longed the survival of treated patients as compared with the controls （17.1 ± 2.01 mo vs 10.1 ±4.5 mo, P = 0.00121）. After treatment, all patients in the study group had positive delayed hypersensitivity and robust systemic cytotoxicity in response to tumor lysate as measured by interferon-y-expression in peripheral blood mononuclear cells using enzyme-linked immunosorbent spot assay. They also displayed increased numbers of CD8-, CD45RO- and CD56-positive cells in the peripheral blood and decreased α-fetoprotein level in the se- rum. CONCLUSION： This new tumor-specific immunotherapy is safe, effective and has a great potential for the treat- ment of tumors.

咳嗽变异性哮喘与典型支气管哮喘患者外周血树突状细胞的变化

目的:计算咳嗽变异性哮喘(CVA)、典型支气管哮喘(CA)患者,其外周血中树突状细胞(DCs)及亚型髓样树突状细胞(m DC)、浆细胞样树突状细胞(p DC)比例和数量的变化,了解咳嗽变异性哮喘与典型支气管哮喘之间是否有类似的免疫学发病机制.方法:选取CA患者20例,CVA患者17例,对照组健康人17例,使用流式细胞术检测和计算CA组与CVA组受试者外周血中DCs及其两个亚群:p DC和m DC所占比例和数量的变化.结果:咳嗽变异性哮喘与典型支气管哮喘患者相比,其外周血树突状细胞及其亚型的数量和比例均无明显差异,咳嗽变异性哮喘、典型支气管哮喘患者较健康对照组,p DC数量及比例的增高明显,m DCs/p DCs的比值降低有统计学差异.结论:咳嗽变异性哮喘与支气管哮喘的发病机制在免疫学上类似,由于p DCs比例和数量的增多引起Th2细胞数量增多,造成Th1/Th2平衡向Th2细胞偏移.

肝脏滤泡树突状细胞肉瘤临床病理特征

目的:探讨肝脏滤泡树突状细胞肉瘤患者的临床病理学特征、诊断、鉴别诊断、治疗方法和预后.方法:以中国解放军南京军区福州总医院收治的3例肝脏滤泡树突状细胞肉瘤患者为对象,分析其临床表现及诊疗经过,并进行随访,同时对送检标本进行病理学检查,结合有关文献探讨该病的临床病理学特点.结果:肿瘤位于肝脏左叶或右叶,呈浸润性生长.组织学上可见梭形细胞肉瘤成分与炎性细胞成分混合存在;局灶见大片凝固性坏死.免疫组织化学染色显示肿瘤细胞呈CD21、CD23、CD35、vimentin强阳性,CD45、EMA呈弱阳性表达,反应性炎症细胞呈CD20、CD79a、CD2、CD3、CD45RO、CD38阳性.例3肿瘤细胞呈EBER、EBV阳性表达.免疫组织化学染色阴性抗体有:CK7、CK8、CK18、CK(H)、CD34、CD117、Dog-1、actin、SMA、caldesmon、desmin、CD10、CD21、CD23、CD35、CD1a、CD2、CD3、CD20、CD45、ALK、CD68、CD163、CD30、CD15、HMB45、S-100.病理诊断为肝脏滤泡树突状细胞肉瘤.结论:肝脏滤泡树突状细胞肉瘤是一种罕见恶性肿瘤,临床与病理上需要与指突状树突细胞肉瘤、肝肉瘤样癌、炎性肌纤维母细胞瘤、胃肠道外间质瘤、组织细胞肉瘤等相鉴别,预后不定.

α-干扰素对慢性髓细胞白血病来源的树突状细胞的免疫调节作用(Ⅱ)

目的 于慢性髓细胞白血病 (CML)来源的树突状细胞 (DCs)的无血清培养基中 ,加入α 干扰素 (IFN α) ,观察IFN α对CML DCs表达Fas/FasL的影响 ,以及体外IFN α合并DCs瘤苗对CML祖细胞的抑制作用。方法 CML DCs的培养基中除加入SCF、GM CSF、TNF α及IL 4外 ,还加入IFN α。培养 10～ 14d ,鉴定细胞免疫表型、Ph1染色体比例和混合淋巴细胞反应 ,流式细胞仪检测细胞表达Fas/FasL比例 ,碘化丙锭 (PI)染色分析细胞凋亡 ,ELISA法测上清液sFas含量 ,甲基纤维素半固体集落培养法鉴定IFN α加DCs瘤苗对CML的CFU GEMM的体外抑制作用。结果 加入IFN α后 ,CML DC免疫表型及刺激T细胞增殖能力均显著改善 ;CML DCsFas的表达上调 ,sFas含量却下降 ,FasL表达阴性 ;PI染色显示亚二倍体峰的数值升高 ,即凋亡比增加。IFN α合并DCs瘤苗组对CML的CFU GEMM体外抑制作用优于IFN α组或DCs瘤苗组。结论 IFN α在改善CML DCs免疫表型及功能的同时 ,亦可通过Fas途径促进Ph1阳性细胞凋亡。IFN α与DCs瘤苗有协同作用 ,体外二者联合应用对CML祖细胞有明显的抑制作用。

子宫颈癌的免疫治疗

近几年有研究以免疫治疗运用于子宫颈恶性肿瘤并取得一定的成效,其中包括检查点抑制剂(immune checkpoint inhibitors)、基于树突状细胞的治疗(dendritic cell-based therapy)、免疫T细胞移植(adoptive T-cell transfer)、肿瘤浸润淋巴细胞免疫细胞移植(tumor infiltrating lymphocytesadoptive cell transfer)、基于细胞毒性T淋巴细胞的免疫疗法(cytotoxic T lymphocytes-based immunotherapy)、使用DC-CIK治疗因子诱导的杀伤细胞的免疫疗法(cytokine-induced killer cell-based immunotherapy)、基于DNA的免疫疗法(DNAbased immunotherapy)。免疫治疗的方法包括结合不同免疫治疗方法,或与放化疗结合使用。这些方法为子宫颈癌的治疗提供了新的希望。然而,免疫治疗的机制和制备过程复杂,且个体化治疗要求高,制备效率低、价格昂贵、周期长等问题亟待解决,需要进一步研究和临床共同努力。

树突状细胞疫苗治疗肝细胞癌的研究进展

肝细胞癌（HCC）为肝脏最常见的恶性肿瘤，早期诊断困难，进展迅速，患者疾病负担较重，预后较差。目前，HCC的主要治疗手段包括手术切除、肝移植、介入治疗、化疗、消融治疗、靶向治疗等，但其改善预后的效果不佳。近年来，树突状细胞在肿瘤免疫治疗中的作用逐渐被认识，并将成为有前景的改善HCC患者预后的治疗方法。本文就树突状细胞疫苗治疗HCC的研究现状进行综述。

无关第三方耐受型树突细胞对小鼠异基因骨髓移植后急性移植物抗宿主病的预防作用

目的探讨无关第三方耐受型树突细胞（tDC）的生物学特性及其对急性移植物抗宿主病（aGVHD）的预防作用。方法用GM—CSF、IL-10和TGF—B1诱导培养D1小鼠骨髓源tDC，用流式细胞术检测细胞表型，用RT—PCR方法检测细胞因子基因表达情况。采用混合淋巴细胞反应法检测其体外对异基因小鼠CD4＋T细胞增殖的影响；将D1小鼠骨髓源tDC按不同剂量分组输注给aGVHD小鼠模型（B6--D2），观察受鼠一般状况、生存率、临床GVHD评分、血清中Thl／Th2细胞因子水平的变化，并以单纯aGVHD模型为对照组进行比较。结果流式细胞术检测结果表明tDC表达低水平的MHCⅡ分子（I—A／I—E）和共刺激分子（CD80、CD86、CD40）。RT—PCR结果表明，tDC低水平表达炎性细胞因子IL-12p40，但高水平表达免疫抑制性因子IL-10、TGF—β、FasL、IDO和精氨酸酶。无关第三方tDC在体外能够有效抑制异基因CD4＋T细胞的增殖，并且这种抑制效应与tDC的剂量有关；小鼠体内实验显示，aGVHD对照组小鼠在移植后18d内全部死亡；而接受无关第三方tDC输注组小鼠aGVHD症状均有所改善，其中输注10。tDC的受鼠有60％能存活至60d以上，无明显的GVHD症状。而输注10^3 tDC的小鼠仅有20％能存活至移植后第60天，输注10^5 tDC的小鼠则在移植后37d内全部死亡。移植21d时，输注10^3、10^4、10^5无关第三方tDC的小鼠血清内IL-10水平分别为（114．23±7．78）、（646．18±212．02）、（121．97±10．47）ng／L，10^4 tDC组显著高于其他组（P〈0．05）。结论经IL—10和TGF-β1诱导的无关第三方tDC在体外能够抑制CD4＋T细胞的增殖，对aGVHD有显著的预防作用，这种作用与tDC剂量和IL-10的高表达有关。过继性输注一定量的无关第三方tDC能显著延长aGVHD小鼠的生存期，有效预防异基因骨髓移植小鼠aGVHD的发生。