Regulation of immune responses by CD1d-restricted natural killer T cells

Natural killer T (NKT) cells are a unique subset of T lymphocytes that share receptor structures and properties with conventional T lymphocytes and natural killer (NK) cells. NKT cells are specific for glycolipid antigens such as the marine sponge-derived agent α-galactosylceramide (α-GalCer) presented by the major histocompatibility complex (MHC) class I-like molcule CD1d. My laboratory has evaluated the function of NKT cells by generating and analyzing CD1d-deficient mice. These studies showed that CD1d expression is required for NKT cell development, but not absolutely necessary for the generation of polarized T helper (Th) cell responses. Further, we have studied the in vivo response of NKT cells toα-GalCer stimulation and the capacity of α-GalCer to modulate innate and adaptive immune responses. Our results revealed that, quickly following administration of α-GalCer, NKT cells expand and produce cytokines, trans-activate a variety of innate and adaptive immune cells, and promote Th2 responses that are capable of suppressing Th1-dominant autoimmunity. Our findings indicate that NKT cells play a regulatory role in the immune response and that specific activation of these cells may be exploited for therapeutic purposes.     

卵巢癌肿瘤标志物研究进展

卵巢癌是妇科三大恶性肿瘤之一,由于疾病早期没有明显的临床症状,发现时多已进展为晚期,且约80%的晚期患者在治疗后3年内复发,因此卵巢癌成为妇科恶性肿瘤中死亡率最高的肿瘤。早期有效的筛查及预后评估机制对卵巢癌的诊断及治疗有重要意义。CA125是监测卵巢癌病程和转归的重要标志物,然而其特异性不足,其他标志物包括HE4、CA199对卵巢癌的诊断和预后也缺乏敏感性和特异性。因此,寻找和鉴定新的分子标志物用于早期筛查和诊断卵巢癌以及评估预后至关重要。本文基于目前卵巢癌生物标志物最新研究进行综述。     

Development and characterization of a novel conjugated methamphetamine vaccine

BackgroundMethamphetamine (METH) addiction is a major public health concern globally with limited management options. The development of a METH vaccine through hapten design has received significant attention as a promising platform for the potential treatment of METH addiction and overdose, however there is yet to be a successful candidate in human trials.Research design and methods: In this study, we developed a novel conjugated METH vaccine using oxidized mannan (a polymannose) as an immunogenic carrier. A METH hapten was synthesized by using amphetamine and conjugated to mannan with a (Lysine-Glycine-Lysine-Glycine-lysine-Glycine-Lysine-Glycine-Lysine-Glycine) (KG)₅ peptide linker.ResultsThe reaction between amphetamine and (KG)₅, oxidation of mannan, and conjugation of amphetamine-(KG)₅ with oxidized mannan were confirmed by color tests, Fourier-transform infrared spectroscopy, gas and liquid chromatography mass spectrometry, thin-layer chromatography, and ultraviolet spectrophotometer. Additionally, the ability of the vaccine to generate antibodies was confirmed in C57BL/6 mice.ConclusionsThe successful development and characterization of the METH-mannan conjugate vaccine, provides a potential therapeutic intervention to curb METH substance use disorders. Each step of vaccine development was characterized to aid in future research on these vaccines, and the immunogenicity shown in the animal models supports future evaluation of the approach. Future studies of the conjugated METH vaccine should evaluate the efficacy in animal models of acute and chronic METH to pave the way for human studies.     

肿瘤抗原冲击致敏的IL-2基因修饰的巨噬细胞治疗肾癌的实验研究

目的 :观察体外肿瘤抗原冲击致敏的白细胞介素 2 (IL 2 )基因修饰的巨噬细胞对肾癌小鼠的治疗效果并探讨其相关的免疫机理。方法 :通过重组腺病毒的介导 ,将IL 2基因转入新鲜分离的小鼠腹腔巨噬细胞 ,经肿瘤抗原冲击致敏后回输治疗原位肾癌小鼠 ,采用 4h5 1 Cr释放法检测脾脏NK和CTL活性。结果 :IL 2基因修饰的巨噬细胞经肿瘤抗原冲击后体内回输可使肾癌小鼠肺转移结节明显减少 ,存活期明显延长 ,40 %肾癌小鼠达到长期存活。治疗后荷瘤小鼠脾脏NK和CTL活性显著提高。结论 :IL 2基因修饰的巨噬细胞经肿瘤抗原冲击后自体回输是治疗肾癌的有效方法。     

幽门螺杆菌重组粘附素rHpaA生物学活性及免疫原性的体外评价

目的：体外评价幽门螺杆菌（Hp)重组粘附素rHpaA的生物学活性及免疫原性,探讨其在Hp疫苗应用中的可行性。方法：流式细胞术和光镜定量计数法检测rHpaA及其抗体对Hp与胃上皮细胞粘附的影响;ELISA法测定Hp感染者血清中抗HpaA抗体,^3H-TDR掺入法测定人外周血淋巴细胞（PBL）对rHpaA的增殖反应,评价rHpaA对体液和细胞免疫的细胞;流式细胞术检测rHpaA对T辅助细胞（Th）表型的选择作用,探讨rHpaA可能的免疫机制。结果rHpaA和抗rHpaA抗血清均能部分阻断Hp与胃上皮细胞系的粘附;ELISA法能检测出Hp阳性患者血清中存在抗HpaA抗体,检测出率与Hp超声破碎抗原（HpSON)包被下的检出率相近（50%vs54%,P＞0．05）;不论培养基中是否加入IL－2,rHpaA均可刺激Hp PBL的增殖（P＜0．05）;Hp阳性或阴性PBL细胞与rHpaA孵育后均能显著提高IL－4阳性T细胞比例（P＜0．05）。结论HpaA是具有免疫原性的Hp菌体成分,且能选择性增加Th2细胞比例,有望成为Hp疫苗研制中一种有效的新抗原。     

重症肝炎免疫治疗与细胞因子、T细胞亚群的调控关系

了解重症肝炎患者经Ｔα１免疫治疗前后外周血中、ｓＩＬ－２Ｒ,ＩＬ－４,ＩＬ－６内毒素及Ｔ细胞亚群的变化。方法采用ＥＬＩＳＡ方法或全成基质偶氮显色法或链菌素亲生物素过氧化酶连接法检测上述指标。结果治疗前各项指标较正常值均有显著变。,后除ｓＩＬ－２Ｒ和ＣＤ＾＋４外,其他指标基本恢复到正常水平。     

Antitumor effects of oral administration of an interferon-inducing pyrimidinone,Bropirimine, on murine renal-cell carcinoma

Bropirimine [2-amino-5-bromo-6-phenyl-4-(3H)-pyrimidinone] is a low-molecular-weight compound that acts as an inducer of interferon in several animal species. Experiments were designed to explore the possibility of using this drug for the treatment of renal-cell carcinoma (RCC). Euthymic BALB/c mice were inoculated with murine RCC (Renca) cells and given graded doses of Bropirimine p.o. for 5 consecutive days beginning on day 1 following tumor inoculation. These mice were killed and tumors were excised on day 21. Bropirimine significantly (P<0.01) inhibited the tumor growth at a daily dose of 1,000 or 2,000 mg/kg. No adverse effect or toxicity was noted at 1,000 mg/kg, and at 2,000 mg/kg there was only a marginal body-weight reduction without any other appreciable side effect. In addition to the inhibition of tumor growth, there was a small yet significant (P<0.05) increase in the duration of survival (in days) in the Bropirimine-treated animals. When the treatment was delayed to begin on day 6 following tumor inoculation, Bropirimine did not suppress tumor growth in euthymic mice, pointing to the importance of the timing of the treatment. In athymic nude BALB/c mice lacking T-cells or T-cell function, Bropirimine also inhibited tumor growth (P<0.01). The antitumor effect of this drug was abolished by pretreatment with anti-asialo GM1 serum, which eliminated natural killer (NK) activity in euthymic mice. In vivo treatment with Bropirimine augmented the cytotoxicity of lymphocytes isolated from the spleens or lungs of the tumor-bearing mice, which were active against Renca and YAC-1 cells in vitro. This activity was NK-cell-dependent as judged on the basis of the results of the in vitro complement-dependent cytotoxicity assay. Since Bropirimine induced interferon (IFN)-/ production, significantly (P<0.05) elevating its serum concentration, and since this drug mimics the effects of IFN-/, it seemed likely that the Bropirimine-induced NK cell augmentation we found was mediated by IFN-/. These results suggest that Bropirimine, a booster of NK activity, may have potential as an adjunct to other therapeutic modalities in the treatment of human RCC.     

Isolation perfusion in extracorporeal circulation with interleukin-2 and lymphokine-activated killer cells in the treatment of in-transit metastases from limb cutaneous melanoma

Background: Therapies of advanced melanoma patients with interleukin-2 (IL-2) and cytotoxic lymphocytes have produced interesting results, but a larger diffusion of these treatments is limited by the severe side effects due to IL-2 systemic infusion. A strictly regional administration of IL-2 and cells by an isolation perfusion (IP) in extracorporeal circulation (ECC) for the treatment of regional melanoma metastases could improve tolerability and efficacy of this specific modality of immunotherapy. Methods: Ten patients were submitted to adoptive immunotherapy with IL-2 and lymphokine-activated killer (LAK) cells by IP in ECC. The schedule of treatment included the first course of a 5-day systemic administration of IL-2 (Proleukin, EuroCetus 9–12 × 10⁶ IU/M²/day continuous infusion); autologous LAK cells were obtained via leukapheresis and after in vitro activation were given (range 8–28 × 10⁹) along with IL-2 (120-2,400 IU/ml of perfusion priming) to the affected limb by IP; IL-2 (9–12×10⁶ IU/m²/day) was also administered by systemic continuous infusion for 5 days starting on the day after IP. Results: All patients concluded the treatment without any major local or systemic toxicities. Clinical responses included one complete and six partial remissions; three patients had stable disease. All patients are alive. Follow-up after IP ranged from 12 to 35 months (median: 22). The analysis of circulating lymphocytes revealed the rapid disappearance of LAK cells, suggesting their extravasation and/or endothelial adhesion in perfused tissues. Conclusions: IP with IL-2 and LAK cells is a new approach for the treatment of in-transit metastases due to cutaneous melanoma. The treatment appears to be feasible and reliable. Further biological and immunological studies should permit amelioration of the present modality of treatment.