483. Oral Adeno-Associated Virus-neu (AAV-neu) Immunotherapy Inhibits the Growth of neu+ Murine Breast Cancer
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Keywords:
HER2/neu
Adeno-associated virus
Oral immunotherapy
Poxviridae
Medical microbiology
Veterinary virology
Orthopoxvirus
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ABSTRACT Recent DNA sequence analysis indicates that rhesus rhadinovirus (RRV) is a member of the lymphotropic gamma-2 herpesvirus family. To determine if RRV is lymphotropic, peripheral blood mononuclear cells from naturally infected monkeys were separated by immunomagnetic bead depletion and analyzed for the presence of RRV by virus isolation and nested PCR. The recovery and consistent detection of RRV in the CD20 + -enriched fraction clearly demonstrates that B lymphocytes are a major site of virus persistence.
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目的
本研究调查6～18岁小麦过敏儿童口服免疫治疗的情况。
方法
本研究开展于2009－2015年，共有4个芬兰儿童过敏诊治单位参与。对于100名小麦过敏的儿童，连续17周每天食用一次全熟的小麦面条，在此过程中小麦蛋白的含量从0.3 mg逐渐升至2 000 mg，并继续维持3个月或9个月。分别于治疗开始前和随访时采集血液样本。
结果
患儿(67%为男性)平均年龄11.6岁(6.1-18.6岁)，其中57人于治疗16月后可以每日食用小麦制品。但100名患儿在治疗过程中有94名出现过敏症状，其中34名为轻度、36名为中度、24名为重度。ω-5-麸朊的特异性IgE在未达治疗剂量的患者中显著升高，并与过敏反应的强度有关。
结论
大部分(57%)小麦过敏的儿童在口服免疫治疗16月后可以食用小麦制品，但100例中，有94例在治疗过程中出现过敏反应，中重度反应达60例。ω-5-麸朊的特异性IgE水平可作为评估小麦耐受量和治疗反应的标志物。
Oral immunotherapy
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In replication of adeno-associated virus type 4 (AAV-4) the helper function may be performed by a non-defective virus from the same group of parvoviruses (Kilham virus). The synthesis of AAV-4 antigen was observed in a pig embryo kidney cell line, SPEV, chronically infected with Kilham virus, strain RV-13, 45--52 passages. A one-day-old SPEV-Kilham culture was infected with AAV-4. The AAV-4 antigen was detected by immunofluorescence at 6, 8, 12, 18 hours, 2, 3, 4, and 5 days after inoculation. During the first 2--4 days after inoculation the AAV-4 antigen was found in the nucleus and perinuclear zone, later in the cytoplasm. A "new" helper virus for AAV-4 replication has been found: simiancytomegalovirus in human embryo fibroblast cell culture permissive for the helper virus. In the systems where AAV-4 replicates, its antigen can be detected in the nucleus and perinuclear zone by IF. AAV-4 did not replicate in a system insensitive to the helper virus or under non-permissive conditions: at the time, the AAV-4 antigen localized only in the cell cytoplasm was detected.
Helper virus
Immunofluorescence
Adeno-associated virus
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Allergen Immunotherapy
Basophil activation
Oral immunotherapy
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The ganglia of rabbits infected with a relatively benign strain of herpesvirus (E-43) and challenged with either of two virulent neurotrophic strains (MP or McKrae) were found to be colonized only by the initial benign infecting strain. Primary infection with the E-43 strain resulted in milder disease when the animals were infected with MP or McKrae strains and also prevented colonization of the ganglion by these strains. Neutralization with anti-glycoprotein C, plaque morphology, cytopathic effects, reconstruction experiments, and restriction endonuclease analysis indicated that the virus recovered from the ganglion was the initial infecting E-43 strain; no traces of the challenging MP and McKrae strains were found. The challenging McKrae strain was shed for several weeks in a few animals, but the virus isolated from the trigeminal ganglia of these animals was the primary infecting E-43 strain. These results suggest that initial infection with a relatively benign strain of herpesvirus may prevent superinfection of the ganglion (but not necessarily the end organ) by highly virulent herpes simplex virus strains and could have significant implications in the consideration of immunization against this disease in humans.
Superinfection
Strain (injury)
Trigeminal ganglion
Simplexvirus
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More than 200 cells were cloned from populations of mammalian cells persistently infected with Japanese encephalitis virus. Only four cloned cultures contained cells that had viral antigen measurable by immunofluorescence and that released infectious virus, yet all clones harbored virus-specific RNA. Superinfection of cloned cells with wild-type Japanese encephalitis virus did not produce cytopathic effects, but resulted in production of viral antigen and infectious virus in formerly nonproducing clones. Cocultivation of nonproducer clone cells with normally permissive cells did not induce virus production, nor did treatment of nonproducer clones with various inhibitors of DNA, RNA, or protein synthesis. It is suggested that the cloning procedure may have selected for a particular subpopulation of cells and that defective virus is also involved in establishment and maintenance of persistent infection.
clone (Java method)
Viral transformation
Superinfection
Viral Interference
Helper virus
Permissiveness
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To study the inhibition of infectious bursal disease virus( IBDV) replication by miRNAs delivered by recombinant avian adeno-associated viral vector,the recombinant avian adeno-associated virus transfer vector for vp2 or vp1 gene-specific miRNAs was built by recombinant DNA technology. Recombinant avian adeno-associated virus was prepared by calcium phosphate method. Virus infestion test revealed that the physical titre reached 5×108TU/ml of purified virus. The total cellular RNA was extracted after the transduction of recombinant virus for detection of expressed miRNAs. After transduction for two days,the DF-1 cells were infected with homologous or heterologous IBDV. Then vp2 gene expression level and virus TCID 50 were tested. Compared with the negative control group,the successful expression of miRNA could significantly inhibit gene amplification on homologous or heterologous virus which was showed by declined titres. Recombinant avian adeno-associated virus could be applied as an effectively and stably expressing vector of miRNA used in develop a new method to prevent infectious bursal disease( IBD).
Infectious bursal disease
Adeno-associated virus
Recombinant virus
Heterologous
Transduction (biophysics)
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Helper virus
Infectious bursal disease
Adeno-associated virus
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RNA interference (RNAi) is a novel antiviral strategy against a variety of virus infections. Infectious bursal disease virus (IBDV) causes an economically important disease in young chickens. This study demonstrated efficient inhibition of IBDV replication by recombinant avian adeno-associated virus (rAAAV)-delivered anti- VP1 and anti- VP2 microRNAs (miRNAs). In the viral vector-transduced cells, sequence-specific miRNA expression was detected by poly(A)-tailed RT-PCR. Reporter assays using a pVP2-EGFP vector showed significant and long-lasting inhibition of VP2–EGFP expression in cells transduced with anti- VP2 miRNA-expressing rAAAV-RFPmiVP2E, but not with the control miRNA-expressing rAAAV-RFPmiVP2con or anti- VP1 miRNA-expressing rAAAV-RFPmiVP1. Semi-quantitative RT-PCR and/or virus titration assays showed a significant inhibitory effect on homologous IBDV replication in cells transduced with rAAAV-RFPmiVP1 or rAAAV-RFPmiVP2E. For two heterologous IBDV isolates, transduction with rAAAV-RFPmiVP1 led to slightly weaker but similar inhibitory effects, whereas transduction with rAAAV-RFPmiVP2E resulted in significantly weaker and different inhibitory effects. These results suggest that rAAAV could act as an efficient vector for miRNA delivery into avian cells and that VP1 is the more suitable target for interfering with IBDV replication using RNAi technology.
Infectious bursal disease
Adeno-associated virus
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