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961.
962.
Gyorffy S Rodriguez-Lecompte JC Woods JP Foley R Kruth S Liaw PC Gauldie J 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2005,19(1):56-63
Canine malignant melanoma (CMM) is a common and aggressive form of cancer in dogs. Established therapeutic approaches such as surgery, chemotherapy, and radiation therapy (RT) have not proven curative. As a coadjuvant of RT and to enhance the antimelanoma immune response, we characterized dendritic cells (DCs) from the bone marrow (BM) of dogs with CMM, ex vivo, for use in therapeutic vaccines. BM mononuclear cells from 3 dogs with melanoma and from 1 healthy dog were cultured for 12 days in media supplemented with recombinant human granulocyte-macrophage colony stimulating factor, stem cell factor, tumor necrosis factor, and Flt-3 ligand. On day 11, DCs were transduced with an adenovirus vector encoding a xenoantigen, human melanoma antigen gp100. Each dog received 3 subcutaneous vaccinations over a 4-month period. Phenotypic analysis of the expanded DC population demonstrated expression of CD11c/CD18 and major histocompatibility complex class II surface markers, and ultrastructural features characteristic of DCs were observed on electron microscopy. On functional analysis, these DCs were able to stimulate allo-reactivity and capture and express gp100. One dog demonstrated antigen-specific cytotoxic T lymphocyte (CTL) activity in peripheral blood lymphocytes. This dog has displayed no clinical signs, either locally or systemically, of recurrent melanoma 48 months after initial DC injection. However, another dog, which was CTL negative, relapsed 22 months after vaccination. Ex vivo DC expansion is feasible for immunotherapy of spontaneous cancers in outbred dogs. 相似文献
963.
【目的】构建猪繁殖与呼吸综合征病毒(PRRSV)GP5基因C3d-P28分子佐剂重组质粒,探明C3d-P28分子佐剂的免疫增强效果。【方法】用大肠杆菌疫苗对健康猪进行免疫激活,提取猪肝脏总RNA,RT-PCR克隆C3d-P28并连接至pUC19载体,构建pUC19-P28.n(2、4、6)串联体,然后分别将P28.n(2、4、6)连接至真核表达载体pcDNA3.1构建成 pcDNA3.1-P28.n(2、4、6)。RT-PCR扩增PRRSV GP5基因,分别定向克隆至pcDNA3.1-P28.n(2、4、6)中,构建pcDNA3.1-GP5-P28.n(2、4、6)重组质粒;提取重组质粒进行双酶切、电泳,同时用重组质粒转染Marc 145细胞,经间接免疫荧光进行蛋白表达检测,并用重组质粒免疫小鼠,通过ELISA试剂盒检测小鼠体内抗体、IL-4和IFN-γ的水平,检验重组质粒的免疫效果。【结果】从猪肝脏中克隆了C3d cDNA,构建了pcDNA3.1-P28.n(2、4、6)串联体,并将PRRSV GP5基因定向克隆至真核表达载体pcDNA3.1-P28.n(2、4、6)中,构建了PRRSV GP5重组质粒(pcDNA3.1-GP5-P28.2、pcDNA3.1-GP5-P28.4、pcDNA3.1-GP5-P28.6);通过检测小鼠血清中抗体、IL-4和IFN-γ的结果显示,抗体效价、IL-4和IFN-γ含量均比对照组高,且以pcDNA3.1-GP5-P28.6组的效果最好,与空白对照及PCDNA3.1-GP5组比较差异均显著(P<0.05)。【结论】构建了猪补体C3d-P28分子佐剂PRRSV GP5重组质粒,该质粒可刺激机体产生较高的体液免疫和细胞免疫水平,补体 C3d-P28分子佐剂能显著增强PRRSV GP5基因的免疫效果。 相似文献
964.
细胞因子与猪瘟病毒E_2基因真核双表达载体的构建及其免疫增强作用 总被引:9,自引:1,他引:9
构建了猪瘟病毒E2 基因与IL 2、IL 3基因的双表达真核表达载体 ,观察其表达水平 ,并对其免疫增强效果进行了观察。结果表明 ,构建了 2种猪瘟病毒E2 基因与白细胞介素 2、3的真核表达质粒pIRSTIL 2 ,pIRSTIL 3。将 2种质粒转染BHK 2 1细胞 ,均可在体外表达E2 抗原和有生物活性的IL 2、IL 3两种质粒能诱导产生CSFV的特异性免疫反应。pIRSTIL 2 ,pIRSTIL 3所诱导的免疫应答反应比使用单表达质粒 pIRST强。试验表明 ,细胞因子与目的基因构建的双表达基因疫苗能有效提高基因疫苗的免疫效果。 相似文献
965.
966.
YAN Lin HE Qi-gai CHEN Huan-chun XIAO Shao-bo WU Mei-zhou LU Jian-qiang HAN Li 《中国农业科学(英文版)》2003,2(8)
Porcine interleukin-2 and porcine interleukin-6 cDNA sequences were cloned into the expressing vectors pET-28a and pGEX-KG respectively. They were expressed in E.coli BL21 (DE3) with high-level production. The gene deleted vaccine of pseudorabies virus Ea strain (TK-/gG-/LacZ+) was mixed with the two different purified recombinant proteins each, or both, with the doses of 2, 5 or 10 μg ml-1. Ten groups of pseudorabies negative antibody swines were immuned twice with tested vaccines with different doses, or control vaccine, respectively. The antibody titers of the test groups were detected by neutralization test, and the daily weight gains of swines were calculated and analyzed statistically. In the study, all the neutralizing antibody titers in test groups were higher than the control group, and the recombinant proteins appeared a dose dependent adjuvant effect. The tested vaccines with 2 μg ml-1 pIL-2 and with 10 μg ml-1 pIL-2/pIL-6 got significant and extremely significant differences, compared with the vaccines without pILs. The difference of the daily weight gain indicated the potential positive influence of pIL-2 and pIL-6 on immune protection. 相似文献
967.
Quercetin, one of the most abundant flavonoids in plants, is discussed with respect to health-promoting effects like antioxidative and anti-inflammatory properties. Although most claims regarding biological effects of flavonoids are based on in vitro and ex vivo studies, the use of flavonoid-containing supplements in humans and companion animals has increased in recent years. Flavonoid-containing supplements are also offered for pet and livestock nutrition. However, any systemic effect of a substance within a living subject depends on its bioavailability. Therefore, the aim of the present study was to gain information on the oral bioavailability of quercetin in horses. Four Icelandic horses with a mean body weight (BW) of 315 ± 25 kg (mean ± standard error [SEM]) were fed a test meal (crimped oats 1 g/kg BW) with the addition of quercetin (20 mg/kg BW). Blood samples were collected directly from the jugular vein before and after ingestion of the test meal for 24 hours, and flavonoid content was analyzed by high-performance liquid chromatography. Quercetin was the main metabolite in plasma with intact flavonol structure after β-glucuronidase/sulfatase treatment of blood samples. The area under the plasma concentration–time curve of quercetin accounted for 88% of total flavonols. Forty-seven percent of the quercetin detected in plasma after ingestion of the test meal was not conjugated. In addition to quercetin, the quercetin derivatives isorhamnetin (methylated) and kaempferol were detected in plasma. Although quercetin is orally bioavailable in horses, similar to other monogastric species, the plasma metabolite pattern differs from those found in species investigated previously (rat, dog, pig, and human). 相似文献
968.
Shijun YAN He YAN Chaolin ZHANG Tongyan WANG Qingyuan YANG Zhe SUN Yan XIAO Feifei TAN Xiangdong LI Kegong TIAN 《农业科学与工程前沿(英文版)》2016,3(1):81
As virulence-determining genes, RR1 and RR2 encode the small subunit and large subunit of viral ribonucleotide reductase (RR) in pseudorabies virus which have been extensively studied in mice. However, their role in pigs has not been adequately investigated. In this study, we deleted RR1 and RR2 genes based on a TK/gE/gI triple gene-deleted pseudorabies virus and tested its efficacy in pigs as a vaccine candidate. The rescued virus showed similar growth properties and plaque size in vitro as its parent strain. In an animal study, the virus could elicit humoral immune responses shown by generation of gB-specific antibodies and virus neutralizing antibodies. However, vaccination could not provide protection against virulent pseudorabies virus challenge since vaccinated pigs showed clinical pseudorabies-specific syndromes. The deficiency in protection may due to the generation of late and low levels of gB antibodies and virus neutralizing antibodies. 相似文献
969.
Immunization of Male Mice with a New Recombinant GnRH Fusion Protein Reduces the Testicular Function 总被引:1,自引:0,他引:1
FANG Fu-gui YANG Ya-ping LIU Ya ZHANG Yun-hai TAO Yong WANG Suo-lu PU Yong ZHANG Xiao-rong 《中国农业科学(英文版)》2009,8(3):380-385
The objective of the present study was to evaluate the effect of an immunogenic maltose-binding protein-gonadotropin releasing hormone (GnRH-6-MBP) using genetic engineering. The synthetic mammalian tandem repeated GnRH hexamer gene was inserted into the expression plasmid pMAL-c2x. Recombinant GnRH-6-MBP protein was over- expressed in E.coli strain BL21. Amylose resin with affinity chromatograph was used to purify target protein. The reactiongenicity of fusion protein was identified by indirect enzyme linked immunosorbent assay (ELISA), and the antigenicity and biological effects of GnRH-6-MBP were tested in mice. In the experiment, 20 male Kunming white mice of 20 d old were randomly divided into treatment and control group. Ten mice were immunized with 100 μg GnRH-6-MBP administered subcutaneously (s.c.) thrice at 2-week intervals with GnRH-6-MBP. Mice were sacrificed after 3 weeks following the booster injection, the testis was removed, weighed and measured, and the histological structure was observed. The reactiongenicity of fusion protein to GnRH antibody was much higher than the control. Active immunization against GnRH-6-MBP reduced remarkably (P 〈 0.01) the length and weight of the testis, and shortened the girth and width of the testis (P 〈 0.05), and suppressed testicular spermatogenesis compared to the control mice. These results indicate that the recombinant GnRH-6-MBP acted as a strong immunogen and caused atrophy of the testis. 相似文献
970.
Despite the range of oral joint supplements available, there has been very limited research into their efficacy. The study aimed to determine the effect of an oral joint supplement on limb kinematics, orthopedic, physiotherapy, and handler evaluation in horses. Supplement S or placebo P was fed to 24 horses for 21 days each in a random order. Horses were evaluated at days 0 (baseline), 21 (after first treatment), and 42 (after second treatment). Assessments included the following: clinical orthopedic evaluation for straight line/lunging circle in walk and trot; high-speed motion capture determined hindlimb kinematics for straight-line trotting; grading of limb range of motion (ROM) and muscle tone based on standardized physiotherapy criteria; handler grading of specific criteria during pasture, groundwork, and ridden exercise. Effect of treatment, sequence, limb, and interactions were investigated using linear-mixed models. S was associated with significantly lower lameness grade in a straight line (P = .001) and circle (P = .010), with individual horses improving up to 2/10 grades over P/baseline. S was associated with significantly improved ROM and muscle tone. Ridden/groundwork scores were significantly higher with S compared to P/baseline. With S, horses were graded significantly higher for “ease of movement” at pasture compared with P/baseline. For horses with hindlimb lameness, S was associated with significantly greater tarsal flexion than baseline (4.2% greater, P < .020) or P (2.7% greater, P < .037). S was associated with less lameness and improved physiotherapy scores, ridden/groundwork scores, and pasture “ease of movement.” Increased midstance tarsal flexion of lame limbs may indicate improved mobility/comfort during peak loading, supporting a positive effect of S. 相似文献