Investigating Virus–Host Interactions in Cultured Primary Honey Bee Cells

Abstract

Honey bee (Apis mellifera) health is impacted by viral infections at the colony, individual bee, and cellular levels. To investigate honey bee antiviral defense mechanisms at the cellular level we further developed the use of cultured primary cells, derived from either larvae or pupae, and demonstrated that these cells could be infected with a panel of viruses, including common honey bee infecting viruses (i.e., sacbrood virus (SBV) and deformed wing virus (DWV)) and an insect model virus, Flock House virus (FHV). Virus abundances were quantified over the course of infection. The production of infectious virions in cultured honey bee pupal cells was demonstrated by determining that naïve cells became infected after the transfer of deformed wing virus or Flock House virus from infected cell cultures. Initial characterization of the honey bee antiviral immune responses at the cellular level indicated that there were virus-specific responses, which included increased expression of bee antiviral protein-1 (GenBank: MF116383) in SBV-infected pupal cells and increased expression of argonaute-2 and dicer-like in FHV-infected hemocytes and pupal cells. Additional studies are required to further elucidate virus-specific honey bee antiviral defense mechanisms. The continued use of cultured primary honey bee cells for studies that involve multiple viruses will address this knowledge gap.

A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis

“…Difficulties are associated with maintaining these cell lines, which may be due to viral infection [ 15 , 17 , 18 ]. In contrast, primary cultured cells have been established from various tissues and organs in the honey bee, and these primary cell cultures are proven to be useful for viral studies and neuronal processes [ 15 , 39 , 40 ]. Although primary cell cultures and ex vivo cultured cells may only be maintained for a limited period of time under healthy conditions, these cells are available for examining gene expression and signaling pathways [ 2 , 5 ].…”

“…The molecular mechanisms underlying insect cellular immunity remains unclear [ 2 ]. Since the in vitro system of insect hemocytes that retains their original characteristics is a requisite for examining the essential processes of cellular immune reactions, attempts to develop hemocyte primary cell cultures have been made in several insects, including the honey bee [ 40 , 63 – 68 ]. Primary cultured honey bee hemocytes could be infected with sacbrood virus and Flock House virus, which replicate intercellularly and peak in abundance 72–96 hours after infection [ 40 ].…”

Procedures and potential pitfalls for constructing a bee-infecting RNA virus clone

“…AmE-711, the published honey bee cell line, has DWV contaminations (18). Although infecting primary cultures of bee tissues are workable host cells for bee viruses (19)(20)(21)(22)(23), lacking virus-free honey bees can curb the decision of developing a primary culture. Moreover, the instabilities of primary cultures can be frustrating and difficult to repeat.…”

Harmlessness, biostimulating and antiviral action of probiotic “Apinormin” for honey bees

“…Сучасне бджільництво зазнає значних збитків під впливом низки негативних чинників, до яких належать і вірусні хвороби. Вони є причиною смертності бджіл, а також знижують резистентність організму до збудників хвороб, несприятливих чинників довкілля, послаблюють силу сімей і, як наслідок, зменшують їхню продуктивність (Brosi et al, 2017;Dynes et al, 2019;Bartlet et al, 2019;Hristov et al, 2020;Luis et al, 2020;Bartlet et al, 2021;Saranchuk et al, 2021;Kovalskyi et al, 2021;McMenamin et al, 2021). До розповсюджених вірусних хвороб личинок медоносних бджіл належить мішечкуватий розплід, який в останні два роки переважає серед інших захворювань розплоду в бджолиних сім'ях в Україні.…”