Lipid-containing Viruses Research Articles

A review of the science and clinical use of alcohol-based hand rubs

Alcohol has a longstanding history as an antiseptic, and the coronavirus disease 2019 (COVID-19) pandemic has sparked a renewed interest in its use as a hand sanitizer. Alcohol works by denaturing protein and rendering cell membranes permeable. It offers excellent germicidal effects against Gram-positive and Gram-negative bacteria, Mycobacterium tuberculosis, fungi, and lipid-containing viruses. However, it is less reliable against non-lipid containing viruses and is ineffective against bacterial and fungal spores. Alcohol-based hand rub (ABHR) usually contains 60–90% isopropanol or ethanol. Additives such as chlorhexidine to complement the action of alcohol and emollients to ameliorate the drying effect of alcohol are often included to improve the formulation of ABHR. In the clinical setting, ABHR provides biocidal activity against multidrug resistant bacteria such as methicillin-resistant Staphylococcus aureus as well as viruses like human coronavirus, severe acute respiratory syndrome coronavirus, and Middle East respiratory syndrome. Moreover, its use is associated with an improved compliance with hand hygiene, which has been shown to translate into better patient outcomes. However, there are cases of intoxications secondary to ingestion of ABHR or adulterated alcohol when resources are diverted away from the normal beverage production to meet the increased need for hand sanitizer during the COVID-19 pandemic. The risk of unintentional topical absorption and fire hazard among healthcare workers is low but should not be ignored. We proposed recommendations to mitigate the risk of ABHR ingestion and poisoning as well as that of fire hazard.

Spectroscopic Studies of Mosquito Iridescent Virus, its Capsid Proteins, Lipids, and DNA

Mosquito iridescent virus (MIV) is an icosahedric lipid-containing virus which affects mosquitos of Aedes, Culex, Culizeta genera. Apart from mosquitos and other insects, iridoviruses cause the mass death of fish and can cause huge losses for industrial fish breedings. The MIV virion consists of a core of the genetic material (double-stranded viral DNA) surrounded by a capsid (icosahedral protein shell) and further encased in a lipid envelope. The aim of the work was to determine the role of MIV virion constituents (lipids, capsid proteins, and viral DNA) in the formation of spectral properties of the whole MIV virions. Measured are UV-Vis absorption, fluorescence, fluorescence excitation, and phosphorescence spectra of MIV virions, their capsid proteins, lipids, and viral DNA dissolved in various buffers. It is shown that the UV absorption of MIV virions is caused by the absorption of all virion constituents such as capsid proteins, lipids, and viral DNA. The fluorescence of MIV virions at room temperature is mainly due to the fluorescence of capsid proteins. The spectra measured at low temperatures make it possible to identify the type of a nucleic acid (DNA or RNA) inside the virion thanks to the fact that the DNA and RNA phosphorescence spectra are radically different.

The Cystovirus phi6 as a simulant for Venezuelan equine encephalitis virus

Innocuous organisms able to simulate the properties of pathogens are necessary so that the behaviour of pathogenic microorganisms in the environment may be understood. This is essential if we are to understand the epidemiology and consequence management of airborne infection, whether natural or the result of a deliberate release. The coliphage MS2 has been widely used as a simulant for mammalian viruses, although it bears little structural or biochemical similarity to the agents of interest. The well-characterised bacteriophage phi6 (host Pseudomonas syringae) is closer in size and complexity to pathogenic viruses, notably the Alphavirus Venezuelan equine encephalitis virus (VEEV) because the virion has surface structures and is bounded by a lipid envelope. Here, we show for phi6 that the assay is simple and precise and that the infectivity is stable in buffers and under conditions used for aerobiology experiments, it is retained after aerosolisation and sampling. Finally, phi6 aerosol was generated into a re-circulating wind tunnel where a stable and acceptable level of viability was maintained. Phi6 may prove a useful simulant for VEEV and possibly for other lipid-containing viruses as well.

New, Closely Related Haloarchaeal Viral Elements with Different Nucleic Acid Types

During the search for haloarchaeal viruses, we isolated and characterized a new pleomorphic lipid-containing virus, Haloarcula hispanica pleomorphic virus 1 (HHPV-1), that infects the halophilic archaeon Haloarcula hispanica. The virus contains a circular double-stranded DNA genome of 8,082 bp in size. The organization of the genome shows remarkable synteny and amino acid sequence similarity to the genome and predicted proteins of the halovirus HRPV-1, a pleomorphic single-stranded DNA virus that infects a halophilic archaeon Halorubrum sp. Analysis of the two halovirus sequences, as well as the entire nucleotide sequence of the 10.8-kb pHK2-plasmid and a 12.6-kb chromosomal region in Haloferax volcanii, allows us to suggest a new group of closely related viruses with genomes of either single-stranded or double-stranded DNA. Currently, closely related viruses are considered to have the same genome type. Our observation clearly contradicts this categorization and indicates that we should reconsider the way we classify viruses. Our results also provide a new example of related viruses where the viral structural proteins have not diverged as much as the proteins associated with genome replication. This result further strengthens the proposal for higher-order classification to be based on virion architecture rather than on genome type or replication mechanism.

Electron Cryomicroscopy Comparison of the Architectures of the Enveloped Bacteriophages ϕ6 and ϕ8

The enveloped dsRNA bacteriophages phi6 and phi8 are the two most distantly related members of the Cystoviridae family. Their structure and function are similar to that of the Reoviridae but their assembly can be conveniently studied in vitro. Electron cryomicroscopy and three-dimensional icosahedral reconstruction were used to determine the structures of the phi6 virion (14 A resolution), phi8 virion (18 A resolution), and phi8 core (8.5 A resolution). Spikes protrude 2 nm from the membrane bilayer in phi6 and 7 nm in phi8. In the phi6 nucleocapsid, 600 copies of P8 and 72 copies of P4 interact with the membrane, whereas in phi8 it is only P4 and 60 copies of a minor protein. The major polymerase complex protein P1 forms a dodecahedral shell from 60 asymmetric dimers in both viruses, but the alpha-helical fold has apparently diverged. These structural differences reflect the different host ranges and entry and assembly mechanisms of the two viruses.

External scaffold of spherical immature poxvirus particles is made of protein trimers, forming a honeycomb lattice

During morphogenesis, poxviruses undergo a remarkable transition from spherical immature forms to brick-shaped infectious particles lacking helical or icosahedral symmetry. In this study, we show that the transitory honeycomb lattice coating the lipoprotein membrane of immature vaccinia virus particles is formed from trimers of a 62-kD protein encoded by the viral D13L gene. Deep-etch electron microscopy demonstrated that anti-D13 antibodies bound to the external protein coat and that lattice fragments were in affinity-purified D13 preparations. Soluble D13 appeared mostly trimeric by gel electrophoresis and ultracentrifugation, which is consistent with structural requirements for a honeycomb. In the presence or absence of other virion proteins, a mutated D13 with one amino acid substitution formed stacks of membrane-unassociated flat sheets that closely resembled the curved honeycombs of immature virions except for the absence of pentagonal facets. A homologous domain that is present in D13 and capsid proteins of certain other lipid-containing viruses support the idea that the developmental stages of poxviruses reflect their evolution from an icosahedral ancestor.

Transmission of infection by gastrointestional endoscopy: May 2001

Transmission of infection by gastrointestional endoscopy: May 2001

Gene XV of Bacteriophage PRD1 Encodes a Lytic Enzyme with Muramidase Activity

Bacteriophage PRD1 is a lipid-containing virus that infects a variety of Gram-negative bacteria, including Escherichia coli. The phage lyses its host by virtue of a virally-encoded lytic enzyme, the synthesis of which has been assigned to gene XV on the basis of complementation analysis and experiments with mutant phages. We report here the cloning of gene XV into an expression plasmid and the purification of its product, protein P15, to near homogeneity. The purified protein P15, identified by N-terminal sequence analysis, showed a strong lytic activity against chloroform-treated Gram-negative cells. No activity against Gram-positive bacterial species could be detected. The pH optimum of the enzyme was between 7.0-8.0. Protein P15 was readily inactivated at temperatures above 4 degrees C, as well as by increasing the ionic strength of the buffers. The analysis of cell wall digests indicated that P15 is a glycosidase that cleaves the beta (1-4) linkage between N-acetylmuramic acid and N-acetylglucosamine, thus displaying muramidase activity.

Differential inactivation of surrogate viruses with merocyanine 540.

Bacteriophages may be useful as surrogates for animal viruses when the virucidal properties of different photosensitizing compounds are initially investigated. We studied photoinactivation of four bacteriophages, phi X174, T7, PRD1, and phi 6, by the dye merocyanine 540 (MC540) (15 micrograms/mL). Merocyanine 540 (MC540) should be most effective with lipid-containing viruses, since it is primarily lipophilic (but also binds to proteins). Two of the phages, PRD1 and phi 6 contain lipid, with only phi 6 having an external lipoprotein envelope. Filtered radiation (450-600 nm) from a 750 W projector was used at 16-100 W/m2. The survival curves of the different viruses clearly demonstrated different levels of sensitivity to photoinactivation by MC540, with phi 6 (Do = 1.5 kJ/m2) being the most sensitive, followed by T7 (21-fold less sensitive). While both PRD1 and phi 6 have lipid components, only phi 6 was photoinactivated by MC540. Thus the internal lipid components of PRD1 were not sufficient to allow photoinactivation by this dye, at fluences up to 300 kJ/m2. For comparison, we also photoinactivated Herpes simplex virus (Do = 0.053 kJ/m2) and found it to be 28-fold more sensitive than phi 6 to photoinactivation by the same concentration of MC540. Thus phi 6 may be used as a surrogate for enveloped human viruses for photoinactivation by lipophilic dyes, but the results may only be useful qualitatively.

Inhibition by albumin of merocyanine 540-mediated photosensitization of platelets and viruses.

The effect of the photosensitizer merocyanine 540 (MC 540) on platelets and on three marker viruses was examined to assess its potential in reducing virus transmission by blood products. The results demonstrated several deleterious effects of MC 540 (4-24 micrograms/mL) on platelet morphology and function in both the absence and presence of light (450-600 nm). Treatment of washed platelets with MC 540 in the dark resulted in a significant release of serotonin in the absence of added agonist, as well as a diminished response to thrombin as measured in vitro. In addition, photosensitization caused spontaneous platelet aggregation and release of 92 percent of the releasable serotonin without the addition of an agonist. Because photo-treatment of blood products is likely to be performed in a protein-rich medium, the influence of albumin on the phototoxic effects on platelets was assessed. Albumin added to the suspension medium at concentrations greater than or equal to 1.0 percent protected the platelets against the effects of MC 540 in the dark, whereas 5-percent albumin was required for protection against the phototoxic effects of MC 540 on the platelet response to thrombin. The antiviral activity of MC 540 and light was examined by using the lipid-containing viruses herpes simplex virus (HSV) and bacteriophages phi 6 and PM2. Of the lipid-enveloped viruses, HSV was 25 times more photosensitive to MC 540 than was phi 6 (15 micrograms/mL). PM2, which has an internal lipid layer, was almost 300 times less sensitive to MC 540 and light than was HSV.(ABSTRACT TRUNCATED AT 250 WORDS)

Structure and assembly of bacteriophage PRD1, an Escherichia coli virus with a membrane

This article describes the structure and assembly of bacteriophage PRD1, a lipid-containing virus able to infect Escherichia coli. This phage, with an approximate diameter of 65 nm, is composed of an outer protein shell surrounding a lipid-protein membrane which, in turn, encloses the nucleic acid. The phage genome consists of a single linear dsDNA molecule of about 15 kb that has a protein covalently linked to each of its 5′ ends. This protein is used as a primer in DNA replication. During assembly membrane proteins are inserted into the host cytoplasmic membrane while major capsid protein multimers are found in the cytoplasm. Capsid multimers, assisted by two nonstructural assembly factors, are capable of translocating the virus-specific membrane resulting in the formation of cytoplasmic empty particles. Subsequent DNA packaging leads to the formation of infectious virus.

Structural study of the lipid-containing bacteriophage PRD1 and its capsid and DNA components by laser Raman spectroscopy

The Raman spectrum of a virus contains the structural signature of each of its molecular components (Thomas, 1987). We report the first Raman spectrum obtained from an intact, lipid-containing virus--the icosahedral bacteriophage PRD1--and show that this spectrum contains characteristic structure markers for the major capsid protein, the packaged double-stranded DNA genome, and the viral membrane which resides between the capsid and DNA. We find that the packaged genome of PRD1 exhibits Raman markers typical of the B-DNA secondary structure. Comparison of the Raman spectrum of the packaged DNA with that of protein-free DNA extracted from the virion shows further that the B-form secondary structure is not significantly perturbed by packaging in the virion. The Raman signature of the PRD1 membrane, monitored within the virion at 4 degrees C, is that of a phospholipid liquid-crystalline phase. The PRD1 capsid, which comprises several hundred copies of the major coat protein P3 (product of viral gene III) and a few copies of minor proteins, incorporates P3 capsomers predominantly in the beta-sheet conformation. The beta-sheet structure of P3 is maintained in the fully assembled PRD1 virion, as well as in the empty capsid. The present results demonstrate the feasibility of obtaining structural information from the three different classes of biomolecules--nucleic acid, protein, and lipid--which constitute a membrane-lined virus particle. Our results also demonstrate that the coat protein and double-stranded DNA components of a lipid-containing bacteriophage share many structural features in common with bacteriophage lacking a lipid membrane.

Trypsin treatment of bovine embryos after in vitro exposure to infectious bovine rhinotracheitis virus or bovine herpesvirus-4

The influence of relative humidity (R.H.) on the survival of the following viruses has been examined; feline herpesvirus (FHV); feline calicivirus (FCV); vesicular exanthema virus (VEV); infectious bovine rhinotracheitis virus (IBRV); parainfluenza 3 virus (PI-3 virus); vesicular stomatitis virus (VSV); equine herpesvirus type 1 (EHV-1), equine arteritis virus (EAV); equine rhinovirus (ERV-1), and African swine fever virus (ASFV). ASFV and PI-3 viruses survived well at all relative humidities when sampled 1 s after formation of an aerosol, but after storage of aerosols for 5 min both viruses were found to be sensitive to high R.H. The other lipid-containing viruses (EAV, VSV, FHV, EHV-1 and IBRV) were unstable when stored as aerosols in moist conditions. The picornavirus ERV-1 was the only virus which survived well at high R.H. but poorly on exposure to dry conditions. The caliciviruses VEV and FCV were sensitive to R.H. in the 30–70% range. When subjected to aeration, all of the lipid-containing viruses which were examined lost infectivity but non-lipid viruses, including bovine adenovirus type 1 (BAdV-1), were stable. The addition of 0.1% peptone reduced losses, probably by protecting against surface inactivation. The significance of these findings in relation to possible control measures for viral respiratory disease is discussed.

Lipid composition of an iridescent virus type 6 (CIV).

The Chilo Iridescent Virus (CIV) is a lipid-containing virus propagated in vitro in choristoneura fumiferana cell cultures. We have analysed the individual lipids of the viral membrane which appeared interesting in their relative amounts and mainly in the high proportion of phosphatidylinositol. This fraction represented about 27 per cent of the phospholipid extract. The lipid composition of the viral membrane was unchanged whether the virus was propagated in vivo in larvae or in vitro in invertebrate cell cultures and was clearly different from that of the hosts.

Enzymatic deoxyglucosylation of ceramides by microsomes of BHK-21 cells: The effect of deoxyglucose treatment and herpesvirus infection

The microsomal fractions of cultured hamster fibroblasts (BHK-21 cells) catalyze the incorporation of glucose from UDPglucose or of deoxyglucose from UDPdeoxyglucose into a reaction mixture with liposomes consisting of ceramide and phosphatidylcholine. The microsomal fractions also catalyze the transfer of glucose from UDPglucose to endogenous acceptors. The specific activity of ceramide deoxyglucoside or ceramide glucoside formation was significantly higher when microsomal preparations obtained from deoxyglucose-treated or herpesvirus-infected BHK-21 cells were used as the glucosyltransferase source. Deoxyglucose was incorporated from UDPdeoxyglucose into hydroxy- and nonhydroxy-fatty acid-containing ceramides at approximately the same rate. Competitive inhibition of deoxyglucosylation of ceramides by UDPglucose suggests that both reactions were catalyzed by the same enzyme, viz. UDPglucose:ceramide glucosyltransferase. This inhibition of glycosphingolipid synthesis may account, in part, for the inhibitory effect of deoxyglucose on lipid-containing viruses.

Influence of the infection with lipid-containing viruses on the metabolism and pools of phospholipid precursors in animal cells.

The influence of infection with three different lipid-containing RNA viruses, Newcastle disease virus, fowl plague virus, and Semliki Forest virus on the phosphatidylcholine precursors of chick embryo cells and of baby hamster kidney (BHK) cells has been measured. In chick embryo cells infection with Newcastle disease virus does not influence the energy charge, or the distribution and absolute pool sizes of the precursors or the choline phosphotransferase activity. In chick embryo cells infected with fowl plague virus the CDP-choline pool increases because of an inhibition of the choline phosphotransferase activity. The phosphorylcholine and CTP pools are smaller in infected cells when compared with mock-infected ones, although the energy charge is not influenced by infection. In chick embryo cells as well as in BHK cells the energy charge is diminished by infection with Semliki Forest virus. Therefore the CTP and phosphorylcholine pools are decreased. The CDP-choline pool in chick embryo cells becomes extremely small after infection with Semliki Forest virus because of a significant stimulation of the choline phosphotransferase. In BHK cells infected with Semliki Forest virus the opposite effect is observed. There are also severe effects on the uptake of the labeled precursors by infection. One and the same virus (Semliki Forest virus) has two completely different effects on the phosphatidylcholine precursors when infecting two different cell types. If one and the same cell type (chick embryo cells) is infected with three different lipid-containing RNA viruses also completely different effects on the phosphatidylcholine precursors were observed. Thus, each virus develops its own strategy to influence the lipid metabolism of the host cell, depending also on the choice of the host. This explains the many disturbing contradictory results described in the literature about the influence of lipid-containing viruses on the lipid metabolism of the host.

Sterol-polyene antibiotic complexation: probe of membrane structure.

Polyene antibiotics are useful tools for studying the role of sterols in biological membranes. The interaction of polyene antibiotics with membrane-bound sterols in artificial membrane systems, prokaryotic and eukaryotic cells, and lipid-containing viruses is reviewed. The pentaene macrolide, filipin, is shown to serve as a probe of phosphatidylcholine-sterol interaction and of the localization of cholesterol in the membrane of mycoplasmas.

The inactivation of lipid-containing viruses in a multistage liquid impinger (May sampler)

The inactivation of lipid-containing viruses in a multistage liquid impinger (May sampler)

The inactivation of lipid-containing viruses in a multistage liquid impinger (May sampler)

The inactivation of lipid-containing viruses in a multistage liquid impinger (May sampler)

Inactivation of Lipid-Containing Viruses by Long-Chain Alcohols

This report describes the inactivation of lipid-containing viruses by several long-chain alcohols. A striking peak in antiviral activity was found for saturated alcohols having chain lengths from 10 to 14 carbons. Viruses having different membrane structure showed different susceptibilities to alcohols having different chain lengths and structural features. Decanol, dodecanol, and tetradecanol readily inactivated herpes simplex virus and the enveloped bacterial virus phi6. The lipid-containing virus PM2 was susceptible to decanol and dodecanol but comparatively unsusceptible to tetradecanol. The branched-chain alcohol phytol, a naturally occurring component of chlorophyll, was active against phi6 and herpes simplex virus but not against PM2. Polyoma virus and the bacteriophage phi23-1-a, which do not contain lipids, were not susceptible to inactivation by any of the alcohols tested. Experiments were also carried out to determine the effects of these compounds on cells. At 0.5 mM, decanol lysed human embryonic lung cells, erythrocytes, and the bacterial hosts for phi6 and PM2. Dodecanol, tetradecanol, and phytol at this concentration were less damaging to cells. At 0.05 mM, none of the alcohols caused observable cytopathic effects on human embryonic lung cells, although several of the alcohols at this concentration were active against herpes simplex virus. Our findings suggest that dodecanol, tetradecanol, and phytol may warrant further studies as potential antiviral agents, particularly for topical application to virus-infected areas of the skin.