Human immunodeficiency virus type 1 envelope-mediated neuropathogenesis: targeted gene delivery by a Sindbis virus expression vector

Abstract

Sindbis virus (SIN) expression vectors offer the opportunity for studying neuropathogenesis because of their distinct neural cell tropism. Here, we demonstrate that a recombinant SIN vector expressing EGFP (SINrep5-EGFP) infected multiple cell types including neural cells from several species relevant to lentivirus pathogenesis with high levels of transgene expression. Infection of human neurons by a recombinant SIN (SINrep5-JRFL) expressing the full-length envelope from a neurovirulent human immunodeficiency virus type 1 (HIV-1) strain (JRFL) caused increased cytotoxicity compared to infection with SINrep5-EGFP (P < 0.001), while no cytotoxicity was observed among infected human astrocytes or monocytoid cells. Both human monocyte-derived macrophages (MDM) (P < 0.01) and astrocytes (P < 0.001) infected with SINrep5-JRFL released soluble neurotoxins in contrast to SINrep5-EGFP or mock-infected cells, although this was most prominent for the astrocytes. Implantation of SINrep5-JRFL into the brains of SCID/NOD mice induced neuroinflammation, neuronal loss, and neurobehavioral changes characteristic of HIV-1 infection, which were not present in SINrep5-EGFP or mock-infected animals. Thus SIN expression vectors represent novel tools for studying in vitro and in vivo HIV-1 neuropathogenesis because of their high levels of transgene expression in specific cell types within the brain.

Aberrant cortical neurogenesis in a pediatric neuroAIDS model: neurotrophic effects of growth hormone

“…1a) with Sindbis vectors expressing EGFP (SIN-EGFP) or the envelope protein from a brainderived neurovirulent HIV-1 strain (SIN-HIVenv) [16], resulted in efficient expression of EGFP or HIV-1 envelope protein depending on the individual viral vector. Moreover, infection by the SIN-HIVenv virus disrupted neurospheres at 24 h after infection (Fig.…”

“…Production of virus stocks The SINrep5 vector system was used including the construction of the SINrep5 vector expressing the enhanced green fluorescent protein (SIN-EGFP) and the envelope protein of brain derived HIV-1 strain JRFL (SIN-HIVenv) [16,17]. The neurovirulent FIV strain used in this study was FIV-Ch [18].…”

West Nile Virus-Induced Neuroinflammation: Glial Infection and Capsid Protein-Mediated Neurovirulence

“…All restriction and other enzymes were obtained from New England Biolabs (Mississauga, Ontario, Canada) or Invitrogen and used according to their specifications. The SINrep5 vector system, the construction of the SINrep5 vector expressing the enhanced green fluorescent protein (EGFP), and the production of recombinant viruses have been described previously (10,63). Briefly, capped in vitro runoff RNA transcripts were generated with the SP6 mMESSAGE mMACHINE kit (Ambion, Inc., Austin, TX) using the linearized pSINrep5-EGFP, pSINrep5-WNV C , and the helper virus construct pDH-BB as templates.…”

“…The WNV capsid protein is considered to play an important role in WNV-induced neuronal death (66). To extend the understanding of the pathogenic properties of the WNV capsid protein, we examined the effects of the expression of the capsid protein in neural cells by cloning the ORF encoding the capsid protein of WNV-NY99 into a neurotropic viral vector derived from Sindbis virus (SINrep5-WNV C ) (10,63). WNV capsid protein was efficiently expressed by the SINrep5-WNV C (Fig.…”

“…The expression of the WNV capsid protein has been implicated in WNV in vivo neurovirulence, based on the implantation of a nonspecific eukaryote vector encoding the WNV capsid protein (66). Using the present Sindbis virus vector expression system, which in addition to neurons infects both astrocytes and cells of macrophage lineage (63), thus resembling the natural infection and tropism by WNV, we studied the effects of WNV capsid protein expression in vivo after striatal implantation of SINrep5-WNV C, SINrep5-EGFP, or supernatants from mocktransduced cultures. Immunohistochemical analyses indicated the activation of both astrocytes (Fig.…”