In vitro model for lytic replication, latency, and transformation of an oncogenic alphaherpesvirus

Abstract

SignificanceMarek’s disease virus (MDV) serves as a versatile small-animal model for herpesvirus-induced oncogenesis. Infection of target cells in vitro was impossible due to the short-lived nature of B and T cells, and analysis of infected cells ex vivo was hampered by their low frequencies. To overcome these limitations, we established an in vitro system that allows infection of target cells with MDV using stimuli that prolong the survival of B and T cells. Our system recapitulates the situation in vivo, including transformation of T cells in vitro. In the future, our system will facilitate the analysis of viral and cellular factors that promote lytic replication, establishment of latency and transformation, processes ultimately resulting in deadly lymphomas in the natural host.

Differential gene expression in chicken primary B cells infected ex vivo with attenuated and very virulent strains of infectious bursal disease virus (IBDV)

“…Chicken primary B cells can be cultured in the presence of chicken CD40L Consistent with previous reports [11,19], we found that when chicken primary B cells were cultured in the presence of soluble chCD40L, the number and viability of the cells was significantly increased compared to when cells were cultured in the absence of chCD40L (Fig. 1).…”

“…Key questions that remain unanswered are the basis for the increased pathogenicity of the vv strain and the mechanism of attenuation of cell culture-adapted strains. However, until recently it has not been possible to culture chicken primary B cells ex vivo because, when they are removed from the BF, they do not survive for long [11]. Consequently, it has not been possible to perform a thorough analysis of the interactions of chicken B cells with different strains of IBDV, and many pathogenesis studies to date have been conducted in vivo, where birds are infected and bursal tissues are harvested at necropsy for downstream analysis of gene expression [12][13][14][15][16][17].…”

“…In 2015, Schermuly et al showed that chCD40L-treated B cells could be infected with Marek's disease virus [11], demonstrating that the cells have the potential for use in the study of the consequences of lymphotropic virus infection. Here we report the successful culture of chicken primary B cells ex vivo in the presence of soluble chCD40L, and provide data demonstrating that these cells can support the replication of an attenuated cell culture-adapted strain of IBDV (D78) and a very virulent strain that does not replicate in non-lymphoid cells (UK661).…”

Differential gene expression in chicken primary B cells infectedex vivowith attenuated and very virulent strains of infectious bursal disease virus (IBDV)

“…Key questions that remain unanswered are the basis for the increased pathogenicity of the vv strain and the mechanism of attenuation of cell-culture adapted strains. However, until recently, it has not been possible to culture chicken primary B cells ex vivo as, when they are removed from the BF, they do not survive for long (11). Consequently, it has not been possible to perform a thorough analysis of the interactions of chicken B cells with different strains of IBDV, and many pathogenesis studies to date have been conducted in vivo , where birds are infected and bursal tissues are harvested at necropsy for downstream analysis of gene expression (12–17).…”

“…The gene encoding chicken CD40 ligand (chCD40L), a molecule responsible for B cell proliferation in vivo , was identified (18) and a soluble fusion protein containing its extracellular domain was subsequently shown to support the proliferation of chicken B cells in culture for up to three weeks (19). In 2015, Schermuly et al showed that chCD40L-treated B cells could be infected with Marek’s disease virus (11), demonstrating that the cells have the potential for studying the consequences of lymphotropic virus infection. Despite these successes, chicken primary B cell cultures have not yet been applied to the study of IBDV.…”