Herpes simplex virus in mice: electron microscopy of neural spread

Herpes simplex virus rapidly infected the trigeminal nerves of mice after intranasal inoculation. Centripetal neural spread was suggested by histologic evidence of encephalitis in the area of attachment of the trigeminal nerve. Furthermore, electron microscopy revealed virus replication primarily within Schwann cells of the trigeminal nerve, and neurons of the gasserian ganglion.     

Activity-dependent transfer of brain-derived neurotrophic factor to postsynaptic neurons

Neurotrophins such as brain-derived neurotrophic factor (BDNF) are thought to be transferred from post- to presynaptic neurons and to be involved in the formation and plasticity of neural circuits. However, direct evidence for a transneuronal transfer of BDNF and its relation to neuronal activity remains elusive. We simultaneously injected complementary DNAs of green fluorescent protein (GFP)-tagged BDNF and red fluorescence protein into the nucleus of single neurons and visualized expression, localization, and transport of BDNF in living neurons. Fluorescent puncta representing BDNF moved in axons in the anterograde direction, though some moved retrogradely, and transferred to postsynaptic neurons in an activity-dependent manner.     

Delayed transneuronal death of substantia nigra neurons prevented by gamma-aminobutyric acid agonist

In an investigation of the mechanism by which brain lesions result in delayed degeneration of neurons remote from the site of injury, neurons within the caudate nucleus of rats were destroyed by local injection of the excitotoxin ibotenic acid. Treatment resulted in the rapid degeneration of the striatonigral pathway including projections containing the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) and delayed transneuronal death of neurons in the substantia nigra pars reticulata. The distribution of nigral cell loss corresponded to the loss of GABAergic terminals. Neuronal death was prevented by long-term intraventricular infusion of the GABA agonist muscimol. Delayed transneuronal degeneration may be produced by neuronal disinhibition consequent to loss of inhibitory inputs. Replacement of inhibitory transmitters by suitable drugs may prevent some forms of delayed neuronal death.     

Herpes simplex virus glycoprotein D: human monoclonal antibody produced by bone marrow cell line

Normal bone marrow cells from a donor positive for herpes simplex virus were transformed with Epstein-Barr virus. The resulting lymphoblastoid cell line has secreted immunoglobulin G1 of the kappa type continuously for 2 years. This immunoglobulin, detected both on the cell surface and in the cytoplasm, reacts with cells infected with herpes simplex virus. It defines an antigen that comigrates with the 55-kilodalton glycoprotein D of herpes simplex virus type 1 and neutralizes the infectivity of herpes simplex viruses 1 and 2.     

TLR3 deficiency in patients with herpes simplex encephalitis

Some Toll and Toll-like receptors (TLRs) provide immunity to experimental infections in animal models, but their contribution to host defense in natural ecosystems is unknown. We report a dominant-negative TLR3 allele in otherwise healthy children with herpes simplex virus 1 (HSV-1) encephalitis. TLR3 is expressed in the central nervous system (CNS), where it is required to control HSV-1, which spreads from the epithelium to the CNS via cranial nerves. TLR3 is also expressed in epithelial and dendritic cells, which apparently use TLR3-independent pathways to prevent further dissemination of HSV-1 and to provide resistance to other pathogens in TLR3-deficient patients. Human TLR3 appears to be redundant in host defense to most microbes but is vital for natural immunity to HSV-1 in the CNS, which suggests that neurotropic viruses have contributed to the evolutionary maintenance of TLR3.     

Noncytotoxic lytic granule-mediated CD8+ T cell inhibition of HSV-1 reactivation from neuronal latency

Reactivation of herpes simplex virus type 1 (HSV-1) from neuronal latency is a common and potentially devastating cause of disease worldwide. CD8+ T cells can completely inhibit HSV reactivation in mice, with interferon-gamma affording a portion of this protection. We found that CD8+ T cell lytic granules are also required for the maintenance of neuronal latency both in vivo and in ex vivo ganglia cultures and that their directed release to the junction with neurons in latently infected ganglia did not induce neuronal apoptosis. Here, we describe a nonlethal mechanism of viral inactivation in which the lytic granule component, granzyme B, degrades the HSV-1 immediate early protein, ICP4, which is essential for further viral gene expression.     

Germline transmission of exogenous genes in the chicken

Difficulties associated with in vitro manipulation and culture of the early chicken embryo have restricted generation of transgenic chickens to approaches that use replication-competent retroviruses. The need to produce transgenic chickens in the absence of replicating virus prompted development of a new method of gene transfer into the chicken. Microinjection of the replication-defective reticuloendotheliosis virus (REV) vector ME111 beneath unincubated chicken embryo blastoderms results in infection of germline stem cells. This vector contains genetic information exogenous to the chicken genome, including both the herpes simplex virus type 1 thymidine kinase gene and the Tn5 neomycin phosphotransferase gene. About 8 percent of male birds hatched from injected embryos contained vector DNA in their semen. All four positive males tested passed vector sequences onto their progeny. Analysis of G1 offspring showed that gonads of G0 male birds were mosaic with respect to insertion of vector provirus. Thus, primordial germ cells present in the unincubated chicken embryo blastoderm are susceptible to infection by defective REV vectors.     

Latency of herpes simplex virus in absence of neutralizing antibody: model for reactivation

Mice inoculated with herpes simplex virus (type 1) by the lip or corneal route and then passively immunized with rabbit antibody to herpes simplex virus developed a latent infection in the trigeminal ganglia within 96 hours. Neutralizing antibody to herpes simplex virus was cleared from the circulation and could not be detected in most of these mice after 2 months. Examination of ganglia from the antibody-negative mice revealed latent virus in over 90 percent of the animals, indicating that serum neutralizing antibody is not necessary to maintain the latent state. When the lips or corneas of these mice were traumatized, viral reactivation occurred in up to 90 percent of the mice, as demonstrated by the appearance of neutralizing antibody. This study provides a model for identifying factors that trigger viral reactivation.     

鸡卵巢传入神经元在脊神经节内的节段性分布

将CB—HRP(霍乱毒素B亚单位结合态的辣根过氧化物酶)溶液注入母鸡卵巢内,逆行追踪研究了分布于卵巢的感觉神经元的定位及其规律性。结果表明:①分布于鸡卵巢,经交感途径传入的感觉神经元胞体位于T_1—L_3节段的脊神经节内,峰值在T_6节段。说明鸡卵巢经交感途径传入的感觉神经元的分布具有相对的集中性和弥散性。②脊神经节内的标记细胞以小细胞居多,推测鸡卵巢内可能含感受损伤性刺激的纤维占绝对优势。     

Vaccinia virus recombinant expressing herpes simplex virus type 1 glycoprotein D prevents latent herpes in mice

In humans, herpes simplex virus causes a primary infection and then often a latent ganglionic infection that persists for life. Because these latent infections can recur periodically, vaccines are needed that can protect against both primary and latent herpes simplex infections. Infectious vaccinia virus recombinants that contain the herpes simplex virus type 1 (HSV-1) glycoprotein D gene under control of defined early or late vaccinia virus promoters were constructed. Tissue culture cells infected with these recombinant viruses synthesized a glycosylated protein that had the same mass (60,000 daltons) as the glycoprotein D produced by HSV-1. Immunization of mice with one of these recombinant viruses by intradermal, subcutaneous, or intraperitoneal routes resulted in the production of antibodies that neutralized HSV-1 and protected the mice against subsequent lethal challenge with HSV-1 or HSV-2. Immunization with the recombinant virus also protected the majority of the mice against the development of a latent HSV-1 infection of the trigeminal ganglia. This is the first demonstration that a genetically engineered vaccine can prevent the development of latency.     

电压门控钠离子通道scn1Laa参与斑马鱼脑神经发育和运动行为调节的研究

电压门控钠离子通道对于脊椎动物脑神经起始、传播动作电位具有重要作用。为了解斑马鱼电压门控钠离子通道基因scn1Laa在脑神经中的作用,通过CRISPR/Cas9基因编辑技术,首次构建了可稳定遗传的生长没有受明显影响的scn1Laa缺陷型（scn1Laa-/-）斑马鱼家系。相比野生型,5 dpf（days post-fertilization,受精后5天）scn1Laa缺陷型斑马鱼兴奋抑制性神经元（氨基丁酸类神经元）表达相对增加,兴奋类神经元（谷氨酸能类神经元）和成熟神经元显著减少,脑部细胞增殖也显著减少。受精后5天和90 天的 scn1Laa缺陷型斑马鱼的运动较同时期野生型斑马鱼更为活跃,受精后90天的 scn1Laa缺陷型斑马鱼的运动具有明显的爆发性。以上结果表明,scn1laa缺失导致兴奋类神经元（谷氨酸能类神经元）以及神经细胞增殖减少,影响脑周围神经放电,导致运动神经调节障碍,出现运动行为异常活跃。即电压门控钠离子通道基因scn1Laa参与斑马鱼脑神经发育和生长,间接参与运动行为调节。同时本文也为进一步探究电压门控钠离子通道在脑神经中的作用奠定基础。     

Viral receptors on isolated murine and human ependymal cells

Viruses that infect ependyma cause ependymitis in humans and hydrocephalus in experimental animals. We report that reovirus type 1 (which induces hydrocephalus in mice) binds to the surface of isolated human and murine ciliated ependymal cells. With the use of recombinant viral clones, the binding property was mapped to the type 1 viral hemagglutinin, which also determines in vivo the affinity of reovirus type 1 for ependyma. Mumps virus, measles virus, parainfluenza type 3, and herpes simplex virus type 1 bind to murine ependyma cells, whereas reovirus type 3, herpes simplex virus type 2, and poliovirus type 2 do not.     

山羊正中神经感觉纤维来源及节段性分布规律的研究—HRP法

本实验采用HRP正中神经断端涂抹法,对五只山羊正中神经感觉纤维的节段性来源,进行了研究。结果表明:正中神经的感觉神经元主要集中于C_7、C_8和Th_1的脊神经节中,其次位于C_6、Th_2的脊神经节中。胞体形态具有多型性,并区分为大、中、小三型。其中以小型细胞为主。     

支配山羊胸腺的神经纤维起源

用ＨＲＰ法研究证明,支配山羊胸腺的初级传入神经来源于双侧Ｃ１～Ｔ５ 脊神经节,以同侧为主;其标记神经元的分布呈双峰型;在双侧结状节也出现少量标记细胞,山羊胸腺的感觉经两侧神经传入,以交感途径占优势;交感节后纤维来自双侧颈前节、颈中节、星状节及Ｔ２～Ｔ５ 椎旁节,其中颈中节和星状节的标记细胞最多;在双侧Ｃ１～２节脊髓腹角和延髓疑核出现标记细胞较多,延髓迷走神经背核、面后核和脊上核出现较少的标记细胞。     

Protection from genital herpes simplex virus type 2 infection by vaccination with cloned type 1 glycoprotein D

Guinea pigs were vaccinated with truncated herpes simplex virus type-1 (HSV-1) glycoprotein D produced in the genetically engineered mammalian cell line gD10.2. Vaccinated animals formed antibodies that neutralized both HSV-1 and herpes simplex virus type 2 (HSV-2) in an in vitro neutralization assay. Vaccinated animals were challenged with HSV-2 by intravaginal infection. Animals that received the immunogen in Freund's complete adjuvant were completely protected from the clinical manifestations of genital HSV-2 infection. Animals that received the immunogen incorporated in alum adjuvants were partly protected from clinical disease; the infections that did develop were significantly less severe than those that occurred in control animals injected with adjuvant alone. The results demonstrate that immunization with a purified viral protein can provide significant protection against primary genital infection by HSV-2 in guinea pigs.     

Rapid diagnosis of herpes simplex virus infections with fluorescent antibody

A fluorescent microscopy technique is described which may prove useful in differentiating clinically similar lesions into lesions of herpes simplex virus etiology and nonherpetic lesions. Herpes simplex virus was isolated from the specimens which yielded positive fluorescence, and no virus was isolated from the specimens which yielded no fluorescence.     

Nonencapsidated infectious DNA of adeno-satellite virus in cells coinfected with herpesvirus

Adeno-associated satellite viruses produce antigen detectable by immunofluorescence but not infectious virus in tissue culture cells coinfected with herpes simplex virus. Analysis of DNA extracts from these infected cells shows that large amounts of infectious satellite virus DNA are produced but not encapsidated in the system. This result indicates that satellite virus may be defective at the maturation step.