Effect of sucrose phosphate and sorbitol on infectivity of enveloped viruses during storage

Howell, C L; Miller, Marjorie J.

doi:10.1128/jcm.18.3.658-662.1983

Cited by 37 publications

(16 citation statements)

References 27 publications

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“…Although sorbitol has been recommended for the transport and holding of CMV and other enveloped viruses, titers may decrease rapidly. Indeed, CMV and VZV infectivity was lost when these viruses were stored in sorbitol, even for short periods of time (33). Virus recovery was significantly better from 0.2 M SP.…”

Section: Liquid Mediamentioning

confidence: 99%

“…Buffered sorbitol and sucrose-based solutions. Sorbitol is a cryoprotectant and thus has been used both for the storage of viruses and as an additive in transport media (33). Among the more frequently isolated viruses are herpesviruses, and, of these, cytomegalovirus (CMV) and VZV are the most labile.…”

Section: Liquid Mediamentioning

confidence: 99%

“…Even so, CMV and VZV recovery after storage is inconsistent, and sorbitol may be toxic for some cell cultures. Sorbitol and 0.2 M sucrosephosphate (SP) were evaluated for suitability as both storage and transport media (33). CMV and VZV recovery was uniformly better from 0.2 M SP than from 70% sorbitol after storage at 20, 4, and -70'C.…”

Section: Liquid Mediamentioning

confidence: 99%

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Transport of viral specimens.

Johnson

1990

Clinical Microbiology Reviews

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Section: Liquid Mediamentioning

confidence: 99%

Section: Liquid Mediamentioning

confidence: 99%

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Transport of viral specimens.

Johnson

1990

Clinical Microbiology Reviews

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“…On the other hand, the remarkable short-term stability of the stocks prepared in SF900II and Express Five, which is advantageously compared with the stability reported for stocks prepared in another commercial serum-free media, 10,12 as well as in UNL-10 supplemented with soybean oil emulsion, could be explained by the presence of sucrose. Sucrose has a direct stabilizing effect on enveloped animal viruses, 41,47,48 including also baculoviruses. 10,12,13 Besides, sucrose could also contribute indirectly to preserve the viral infectivity through the stabilization of the lipid emulsion structure.…”

Section: Discussionmentioning

confidence: 99%

Reduction of the infectivity of baculovirus stocks frozen at ultra‐low temperature in serum‐free media: The role of lipid emulsions

Eberhardt

Gioria

Micheloud

et al. 2016

Biotechnology Progress

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The infectivity of stocks of baculoviruses produced in serum-free media is sensitive to freezing at ultra-low temperatures. The objective of this work was to elucidate the causes of such sensitivity, using as a model the freezing of stocks of Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV), a baculovirus widely employed as biological insecticide. Titers of supernatants of cell cultures infected with AgMNPV in four different serum-free media supplemented with lipid emulsions were reduced by 50 to 90% after six months freezing. By using a full factorial experiment, freezing and lipid emulsion, as well as the interaction between them, were identified as the main factors reducing the viral titer. The virucidal effect of the lipid emulsion was reproduced by one of their components, the surfactant Polysorbate 80. Damaged viral envelopes were observed by transmission electron microscopy in most particles frozen in a medium supplemented with lipid emulsion or Polysorbate 80. Additionally, Polysorbate 80 also affected the infectivity of AgMNPV stocks that were incubated at 27°C. The identification of the roles played by the lipid emulsion and Polysorbate 80 is not only a contribution to the understanding of the mechanisms underlying the inactivation of baculovirus stocks produced in serum-free media during storage at ultra-low temperature, but is also an input for the rational development of new procedures aimed at improving both the preservation of baculovirus stocks and the composition of culture media for the production of baculovirus-based bioproducts in insect cells. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1559-1569, 2016.

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“…Viral particles were concentrated by ultracentrifugation in an SW28 rotor (Beckman Coulter, Krefeld, Germany) at 22,000 rpm for 1 h at 4°C. Pellets were resuspended in a 1:1 mixture of medium and 0.2 M sucrose phosphate (84). Aliquots were stored at Ϫ80°C.…”

Section: Methodsmentioning

confidence: 99%

Interaction of Human Cytomegalovirus Tegument Proteins ppUL35 and ppUL35A with Sorting Nexin 5 Regulates Glycoprotein B (gpUL55) Localization

Maschkowitz

Gärtner

Hofmann-Winkler

et al. 2018

J Virol

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Human cytomegalovirus (HCMV) is a widespread human pathogen that causes asymptomatic infection in healthy individuals but poses a serious threat to immunocompromised patients. During the late phase of HCMV infection, the viral capsid is transported to the cytoplasmic viral assembly center (cVAC), where it is enclosed by the tegument protein layer and the viral envelope. The cVAC consists of circularly arranged vesicles from the -Golgi and endosomal networks. The HCMV gene UL35 encodes ppUL35 and its shorter form, ppUL35A. We have previously shown that the UL35 gene is involved in HCMV assembly, but it is unknown how UL35 proteins regulate viral assembly. Here we show that sorting nexin 5 (SNX5), a component of the retromer and part of the retrograde transport pathway, interacts with UL35 proteins. Expression of wild-type proteins but not mutants defective in SNX5 binding resulted in the cellular redistribution of the cation-independent mannose-6-phosphate receptor (CI-M6PR), indicating that UL35 proteins bind and negatively regulate SNX5 to modulate cellular transport pathways. Furthermore, binding of UL35 proteins to SNX5 was required for efficient viral replication and for transport of the most abundant HCMV glycoprotein B (gB; gpUL55) to the cVAC. These results indicate that ppUL35 and ppUL35A control the localization of the essential gB through the regulation of a retrograde transport pathway. Thus, this work is the first to define a molecular interaction between a tegument protein and a vesicular transport factor to regulate glycoprotein localization. Human cytomegalovirus is ubiquitously present in the healthy population, but reactivation or reinfection can cause serious, life-threatening infections in immunocompromised patients. For completion of its lytic cycle, human cytomegalovirus induces formation of an assembly center where mature virus particles are formed from multiple viral proteins. Viral glycoproteins use separate vesicular pathways for transport to the assembly center, which are incompletely understood. Our research identified a viral structural protein which affects the localization of one of the major glycoproteins. We could link this change in glycoprotein localization to an interaction of the structural protein with a cellular protein involved in regulation of vesicle transport. This increases our understanding of how the virus intersects into cellular regulatory pathways to enhance its own replication.

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Effect of sucrose phosphate and sorbitol on infectivity of enveloped viruses during storage

Cited by 37 publications

References 27 publications

Transport of viral specimens.

Transport of viral specimens.

Reduction of the infectivity of baculovirus stocks frozen at ultra‐low temperature in serum‐free media: The role of lipid emulsions

Interaction of Human Cytomegalovirus Tegument Proteins ppUL35 and ppUL35A with Sorting Nexin 5 Regulates Glycoprotein B (gpUL55) Localization

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