A prime-boost concept using a T-cell epitope-driven DNA vaccine followed by a whole virus vaccine effectively protected pigs in the pandemic H1N1 pig challenge model

Influenza A virus (IAV) vaccines in pigs generally provide homosubtypic protection but fail to prevent heterologous infections. In this pilot study, the efficacy of an intradermal pDNA vaccine composed of conserved SLA class I and class II T cell epitopes (EPITOPE) against a homosubtypic challenge was compared to an intramuscular commercial inactivated whole virus vaccine (INACT) and a heterologous prime boost approach using both vaccines. Thirty-nine IAV-free, 3-week-old pigs were randomly assigned to one of five groups including NEG-CONTROL (unvaccinated, sham-challenged), INACT-INACT-IAV (vaccinated with FluSure XP® at 4 and 7 weeks, pH1N1 challenged), EPITOPE-INACT-IAV (vaccinated with PigMatrix EDV at 4 and FluSure XP® at 7 weeks, pH1N1 challenged), EPITOPE-EPITOPE-IAV (vaccinated with PigMatrix EDV at 4 and 7 weeks, pH1N1 challenged), and a POS-CONTROL group (unvaccinated, pH1N1 challenged). The challenge was done at 9 weeks of age and pigs were necropsied at day post challenge (dpc) 5. At the time of challenge, all INACT-INACT-IAV pigs, and by dpc 5 all EPITOPE-INACT-IAV pigs were IAV seropositive. IFNγ secreting cells, recognizing vaccine epitope-specific peptides and pH1N1 challenge virus were highest in the EPITOPE-INACT-IAV pigs at challenge. Macroscopic lung lesion scores were reduced in all EPITOPE-INACT-IAV pigs while INACT-INACT-IAV pigs exhibited a bimodal distribution of low and high scores akin to naïve challenged animals. No IAV antigen in lung tissues was detected at necropsy in the EPITOPE-INACT-IAV group, which was similar to naïve unchallenged pigs and different from all other challenged groups. Results suggest that the heterologous prime boost approach using an epitope-driven DNA vaccine followed by an inactivated vaccine was effective against a homosubtypic challenge, and further exploration of this vaccine approach as a practical control measure against heterosubtypic IAV infections is warranted.     

MBP对PBMC产生IFN-γ和NO的影响

目的探讨T细胞非特异性活化在CNS脱髓鞘性疾病中的作用。方法分离实验性自身免疫性脑脊髓炎(EAE)易感性BALB/c小鼠外周血单个核细胞(PBMC),采用体外细胞培养方法在体外与碱性髓鞘蛋白(MBP)共培养,测定培养上清液中IFN-γ、NO水平。结果经MBP刺激的PBMC产生IFN-γ[(43.83±6.06)pg/mL]和NO[(180.76±20.75)μmol/L]明显增加,与对照组产生的IFN-γ[(28.52±2.18)pg/mL]和NO[(95.61±13.09)μmol/L]相比差异有统计学意义(P<0.01)。结论在CNS脱髓鞘性疾病发病过程中,活化的T细胞、单核细胞等分泌致炎细胞因子和其他有害物质增多。     

The effect of interleukin-1 on iron metabolism in rats

The effect of interleukin-1 on iron metabolism in rats was evaluated. Plasma iron decreased from 184 +/- 16 micrograms/dl (mean +/- SE) to 24 +/- 12 at 6 hours after interleukin-1 intramuscular administration in non-fasting rats and 109 +/- 6 micrograms/dl to 12 +/- 1 micrograms/dl in fasting rats, which was significantly lower than in control rats. Ferrokinetic studies showed a more rapid disappearance rate and lower iron turnover in interleukin-1-injected rats. The release of iron from the mononuclear phagocyte system to plasma was studied at 3 h after interleukin-1 administration. Although the percent of radioactivity in plasma of the total injected dose was 3.2 +/- 0.6% in interleukin-1, which was significantly lower than in the control rats (5.4 +/- 0.6%) at 9 h after intravenous injection of 59Fe chondroitin ferrous sulfate, there was no difference between the amount of 59Fe released from the mononuclear phagocyte system over the first 9 h in interleukin-1 and control rats. These data appear to imply that iron release is unimpaired but that, for some reason, there is an enhanced rate of clearance of the 59Fe once it has been released from the mononuclear phagocyte system into the plasma.     

脐血单核细胞对VD大鼠认知功能及脑组织BDNF的影响

目的观察颈内动脉输注脐血单核细胞(Human cord blood mononuclear cells,HCMNCs)对血管性痴呆(Vascular dementia,VD)大鼠认知功能及脑组织脑源性神经营养因子(Brain-derivedneurotrophicfactor,BDNF)含量的影响。方法改良Pulsinellis四血管阻断法建立VD大鼠模型;体外分离HCMNCs,术后24h颈内动脉输注数量为3×106/0.5ml的BrdU标记细胞;利用穿梭箱系统和ELISA法检测注射HCMNCs后2、4、8周VD大鼠学习记忆能力以及脑组织BDNF含量的变化。结果模型组大鼠主动回避反应比率明显低于对照组(P<0.01),治疗组较模型组显著提高(P<0.01)。术后2周模型组大鼠脑组织BDNF含量较对照组明显增高(P<0.01),4周时达到高峰(P<0.01),8周时则明显下降,与2周时相比有显著性差异(P<0.05);颈内动脉输注HCMNCs后治疗组大鼠脑组织BDNF含量较模型组显著升高(P<0.01),4周时最高(P<0.01),8周时略有下降,但仍维持在较高水平,与4周时相比无显著性差异(P>0.05)。结论颈内动脉输注HCMNCs可显著改善VD大鼠学习记忆能力,增加VD大鼠脑组织BDNF含量,具有脑保护作用。     

激素敏感型肾病单个核细胞培养上清液对大鼠肾小球多阴离子及超微结构的影响

用刀豆蛋白Ａ刺激激素敏感型肾病单个核细胞，并将其培养上清液灌注于大鼠肾内。结合电镜观察，用图像分析仪及体视学方法分别对其肾组织胶体铁染色切片和电镜照片上的足突和滤过裂隙的平均宽度进行测定。结果表明：该病极期上清液灌注的鼠肾小球多阴离子明显减少，足突肿胀、融合，滤过裂隙变窄；而该病缓解组和正常对照组则为阴性。说明该病单个核细胞培养上清液可引起鼠肾小球微小病变型肾病综合征的病变，并揭示该变化可能是由其上清液中一种异常的Ｔ淋巴细胞因子所致。     

单核吞噬细胞应答伤寒沙门菌的实验研究

目的 研究单核吞噬细胞应答伤寒沙门菌的机理。方法 采用体外培养的PMA分化的THP－1细胞，检测其内化和杀伤胞内伤寒沙门菌的活性，以及伤寒沙门菌诱导PMA分化的THP－1细胞产生TNF－α和IL－12的情况。结果 PMA分化的THP－1细胞内化和杀伤Vi阴性伤寒沙门菌的活性明显高于内化和杀伤Vi阳性伤寒沙门菌的活性。IFN－γ明显中此活性。伤寒沙门菌诱导PMA分化的THP－1细胞可以产生一定量的TNF－α和IL－12，然而，IFN－γ活化的PMA分化的HTP－1细胞产生TNF－α和IL－12的量明显增加。结论 伤寒沙门菌在单核吞噬细胞中的存活可能与Vi抗原的存在有关；IFN－γ在调节机体细胞免疫，防循伤寒沙门菌感染中起着重要的作用。     

Induction of type II collagen-specific antibody production in blood lymphocyte cultures of rhesus monkeys (Macaca mulatta) with collagen-induced arthritis using the immobilized native antigen.

Peripheral blood mononuclear cells (PBMC) from Rhesus monkeys previously immunized with bovine type II collagen to induce arthritis were cultured with the same antigen. Because the native protein is poorly soluble in culture medium a heating step is often used. The antigen in this form induced PBMC proliferation, but epitopes for the induction of antibody production and arthritis were lost. To keep the native protein intact it was coated on affigel beads. With the immobilized antigen specific antibody production could be induced.     

用Fura-2测定周围血单个核细胞内游离钙浓度

本文用Fura-2测定了周围血单个核细胞内游离钙浓度，并对有关实验条件进行了探讨。对比研究表明，血标本在4℃保存4小时对测定结果无显著影响，测定标本中单个核细胞数以10~5～10~6/ml为宜。认为测定单个核细胞内游离钙浓度对判断单个核细胞的功能状况有一定帮助。     

脂肪乳剂对健康人外周血单个核细胞磷脂脂肪酸组成的影响

目的探讨脂肪乳剂对健康人免疫细胞脂肪酸组成的影响。方法采用毛细管气相色谱法检测8名健康志愿者连续7天外周静脉输注脂肪乳剂(20%intralipid)后,外周血单个核细胞磷脂脂肪酸谱的变化。结果静脉输注脂肪乳剂对健康人外周血单个核细胞数量、肝功能和血脂无影响,软脂酸(C16∶0)、油酸(C18∶1N9)比例在单个核细胞磷脂酰乙醇胺(PE)中明显增加(P<0.05,P<0.01);硬脂酸(C18∶0)减少(P<0.05);亚麻酸(C18∶3N3)、软油酸(C16∶1N7)在磷脂酰胆碱(PC)中明显增加(P<0.05),而其余脂肪酸均未发生明显改变。饱和脂肪酸与不饱和脂肪酸的比值(S/U)和脂肪酸的不饱和指数(UI)在单个核细胞PE、PC中均保持不变。结论脂肪乳剂不可能通过影响免疫细胞磷脂脂肪酸组成的途径对免疫功能产生影响。