Induction and mode of action of the viral stress-inducible murine proteins, P56 and P54

Mammalian cells respond to virus infection or other viral stresses, such as double-stranded (ds) RNA and interferons (IFN), by robust and rapid induction of viral stress-inducible proteins. The induction and actions of one such protein, the human P56, have been extensively studied. However, little is known about the distantly related mouse proteins, MuP56 and MuP54. Here, we report that, in mouse cells, they could be induced by IFN, dsRNA or Sendai virus infection. MuP56 and MuP54 inhibited protein synthesis in vitro by binding to the "c", but not the "e", subunit of the translation initiation factor, eIF-3. The N-terminal region of the MuP54 was sufficient for inhibiting translation, but it and the corresponding region of MuP56 bound to two different regions of eIF3c. Thus, members of the human and murine P56 family have similar but non-identical functions.     

膜融合脂质体为载体的黑色素瘤疫苗的研究

 目的制备膜融合脂质体为载体的黑色素瘤疫苗，并评价免疫小鼠所产生的细胞免疫和体液免疫。方法从B₁₆黑色素瘤中提取混合蛋白质作为抗原，将其包封在脂质体中并与仙台病毒融合形成的膜融合脂质体，制成疫苗。考察膜融合脂质体的形态及粒径分布，测定小鼠免疫后的细胞毒T淋巴细胞(CTL)反应活性及血浆中总的IgG的浓度。结果以普通脂质体为载体或游离的蛋白质制备的疫苗只能产生体液免疫反应而不能诱发细胞毒T淋巴细胞反应；以膜融合脂质体为载体的疫苗既可产生体液免疫又可诱发强烈的CTL反应(P＜0.001)，并能抑制肿瘤细胞的生长。结论膜融合脂质体是一种有效的肿瘤疫苗载体。     

Intratumoral and s.c. injection of inactivated hemagglutinating virus of Japan envelope (GEN0101) in metastatic castration‐resistant prostate cancer

Inactivated hemagglutinating virus of Japan envelope (HVJ‐E) has an antitumor effect and tumor immunity. We undertook an open‐label, phase I, dose‐escalation study in patients with castration‐resistant prostate cancer (CRPC) to determine the safety and efficacy of intratumoral and s.c. injection of HVJ‐E (GEN0101). Patients with CRPC, who were resistant to or unable to receive standard of care, were included. GEN0101 was injected directly into the prostate and s.c. in two 28‐day treatment cycles. The primary end‐points were to evaluate the safety and tolerability of GEN0101 and determine its recommended dose. The secondary end‐points were to analyze the antitumor effect and tumor immunity. Three patients received 30 000 mNAU GEN0101 and 6 received 60 000 mNAU. There was no dose‐limiting toxicity, and the recommended dose of GEN0101 was defined as 60 000 mNAU. Radiographically, 1 patient had stable disease and 2 had progressive disease in the low‐dose group, whereas 5 patients had stable disease and 1 had progressive disease in the high‐dose group. Three patients in the high‐dose group showed reduction in lymph node metastasis. Prostate‐specific antigen increase rates in the high‐dose group were suppressed more than those in the low‐dose group. Natural killer cell activity was enhanced in 2 patients of the low‐dose group and in 5 patients in the high‐dose group. In conclusion, intratumoral and s.c. injections of GEN0101 were well‐tolerated and feasible to use. The study is registered with the UMIN Clinical Trials Registry (no. UMIN000017092).     

仙台病毒F基因的克隆及原核表达

目的利用原核表达系统表达仙台病毒(Sendai Virus)F蛋白主要抗原片段FP(S),并对表达产物进行免疫学初步研究。方法根据GenBank公布的仙台病毒F蛋白(gi:9627219)的基因序列设计特异性引物,通过RT-PCR扩增出F基因的主要抗原片段FP(S),插入pMD-18-T载体中,鉴定正确后克隆入pQE31原核表达载体中,将鉴定正确的pQE31-FP(S)转化大肠埃希菌M15,IPTG诱导表达,对大肠埃希菌裂解物进行SDS-PAGE和Western-blot验证。结果大肠埃希菌表达的FP(S)相对分子质量约26×103,与预期相符;能与SeV阳性血清发生特异性反应,出现单一条带。结论原核表达的FP(S)蛋白有良好的抗原性,为检测仙台病毒抗体的ELISA检测方法的研究奠定了基础。     

Follow-up studies of long-term survivors after hepatic portoenterostomy for “noncorrectable” biliary atresia

Fourteen patients with “noncorrectable” biliary atresia are living without jaundice for more than 2 yr after hepatic portoenterostomy or its modification. Retardation of physical growth was observed in one of them, and mental retardation in another, both of which seemed irrelevant to biliary atresia. Serial tests for liver function after operation revealed early recovery of serum bilirubin, transaminase, and turbidity, and delayed improvement of alkaline phosphatase. Postoperative needle biopsy of the liver disclosed that changes in hepatic parenchyma and ductular proliferation were rapidly improved after successful operation. Improvement of fibrosis of the liver was delayed, and it was not satis-factory in patients whose preoperative changes in the liver were severe or in whom ascending cholangitis had been a frequent complication. Histologic features of hepatic cirrhosis were observed in the liver in three cases, in two of which there had been frequent episodes of cholangitis. Only one of these showed clinical signs of portal hypertension.Functional and morphologic cure can be achieved in “noncorrectable” biliary atresia by hepatic portoenterostomy or its modifications, although varying degree of hepatic fibrosis may remain according to severity of preoperative changes of the liver and postoperative complication of ascending cholangitis.     

Cytotoxic T lymphocytes specific for vesicular stomatitis virus recognize the major surface glycoprotein of VSV

The major surface glycoprotein (G) of vesicular stomatitis virus (VSV) was purified and incorporated into lipid vesicles containing the purified hemagglutinin-neuraminidase (HN) and fusion (F) glycoproteins of Sendai virus. These lipid vesicles were then used to modify and render target cells susceptible to lysis by anti-VSV cytotoxic T lymphocytes (CTLs). This result provides direct evidence that the G protein is a target antigen of anti-VSV CTLs.     

Incorporation of orally administered glucose-U-14C and fructose-U-14C into the triglyceride of liver, plasma, and adipose tissue of rats

Male rats, fasted for 5–6 hr, were given glucose-U-¹⁴C or fructose-U-¹⁴C, with their respective carrier, by intragastric instillation. Sequential radioactivity in plasma carbohydrates and in the triglyceride of the liver, plasma, and adipose tissue and plasma immunoreactive insulin and free fatty acids were measured. The validity of taking the triglyceride labeling rate as the triglyceride synthesis rate was tested by measuring the metabolic activity of endogenous glucose. Two or three times greater radioactivity was found in the liver and plasma triglycerides after fructose than after glucose while the reverse was true for adipose tissue. The greater radioactivity in triglyceride of the liver and plasma after fructose was mainly due to triglyceride-glycerol radioactivity. The greater radioactivity in adipose tissue triglyceride was due to the radioactivity of both triglyceride-fatty acids and triglyceride-glycerol. Three hours after glucose, 92% of the total radioactivity in triglyceride was in adipose tissue and 8% was in the liver. However, 3 hr after fructose, 57% of the radioactivity was in adipose tissue and 42% was in the liver. Daily repetition of such a pattern of fructose handling may lead to abnormal metabolism of endogenous triglyceride.     

Persistent inflammation and hyperresponsiveness following viral rhinosinusitis

OBJECTIVES: To develop a murine model of viral rhinosinusitis. STUDY DESIGN: Randomized, controlled, animal model. METHODS: Mice were intranasally inoculated with Sendai virus (SeV) or ultraviolet (UV)-inactivated virus. On days 3 and 10 postinfection, nasal lavage fluid was obtained for viral culture. On days 4, 10, and 38 postinfection, sinus mucosa was harvested and analyzed by flow cytometry for CD3-, CD4-, CD8-, CD25-, CD11b-, CCR3-, and GR1-positive cells. Nasal hyperresponsiveness to histamine challenge was measured on days 8 and 36 postinoculation. RESULTS: On day 3, viral cultures were positive from all SeV-inoculated mice but from none of the UV-inactivated mice (P相似文献