Cell Lines for Honey Bee Virus Research

Abstract

With ongoing colony losses driven in part by the Varroa mite and the associated exacerbation of the virus load, there is an urgent need to protect honey bees (Apis mellifera) from fatal levels of virus infection and from the non-target effects of insecticides used in agricultural settings. A continuously replicating cell line derived from the honey bee would provide a valuable tool for the study of molecular mechanisms of virus-host interaction, for the screening of antiviral agents for potential use within the hive, and for the assessment of the risk of current and candidate insecticides to the honey bee. However, the establishment of a continuously replicating honey bee cell line has proved challenging. Here, we provide an overview of attempts to establish primary and continuously replicating hymenopteran cell lines, methods (including recent results) of establishing honey bee cell lines, challenges associated with the presence of latent viruses (especially Deformed wing virus) in established cell lines and methods to establish virus-free cell lines. We also describe the potential use of honey bee cell lines in conjunction with infectious clones of honey bee viruses for examination of fundamental virology.

Alternatives for obtaining a continuous cell line from Apis mellifera

“…The AmE-711 is originated from honeybee eggs and is a strain that has not undergone an immortalization process, and presents characteristics of a finite lineage, such as being ,is limited to a few passages, low cell proliferation and is the one which the scientific society does not have commercial access. This cell line proved difficult to maintain and crashed in 2015 but have been recovered and adapted to a commercially medium (CARRILLO-TRIPP et al, 2016;BEAUREPAIRE et al, 2020;GUO et al, 2020). The MyN9 is originated from honeybee larvae and is the only report of cultivation of this species that was induced to immortalization.…”

“…As a way to overcome this situation of the unavailability of Apis mellifera cell lines, studies with viruses, currently carried out, use primary or subcultivated cultures from different tissues and at different stages of development of this species (GENERSCH et al, 2013;BEAUREPAIRE et al, 2020). So far, several primary cultures with 6 to 135 days have been reported, originating from eggs, pupae, intestines, nervous tissue, among others (POPPINGA et al, 2012;JU & GHIL, 2015;GUO et al, 2020).…”

“…Telomerases are ribonucleoproteins that have an RNA component (hTERC) and a catalytic component (hTERT) (HARRINGTON et al, 1997), whose function is to restore base pairs of DNAs lost from telomeres during cell division (MASON et al, 2011). The length of the chromosomes is restored after each cell division in cells that have active telomerases (ZVEREVA et al, 2010).…”

Investigating Virus–Host Interactions in Cultured Primary Honey Bee Cells

“…In spite of their utility and importance, robust cell culture systems are not available for many non-model organisms, including honey bees ( Apis mellifera ). While approximately 1000 cell lines have been derived from insect tissues, including 19 hymenopteran cell lines (the insect order that includes bees, ants, and wasps), only one immortalized honey bee cell line, AmE-711, has been established [ 8 , 9 , 10 ].…”

DWV Infection in vitro Using Honey Bee Pupal Tissue

“…The DWV can propagate in honey bee larvae and pupae by viral injection ( Lamp et al, 2016 ; Gusachenko et al, 2020 ; Ryabov et al, 2020 ); however, this in vivo system does not allow us to study the underlying mechanisms of viral infection and replication. Virus-free immortalized honey bee cell lines provide the best resource to study viruses and other pathogens ( Guo et al, 2020 ); however, these cell lines are not yet available. To solve this problem, we developed an in vitro system to reconstitute the DWV infection using cultured honey bee pupal tissue and RNA-dependent RNA polymerase (RdRP) as a marker.…”