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PubMed2002

Production of core and virus-like particles with baculovirus infected insect cells.

In this paper the fundamental aspects of process development for the production of core and virus-like particles with baculovirus infected insect cells are reviewed. The issues addressed include: particle formation and monomer composition, chemical and physical conditions for optimal cell growth, baculovirus replication and product expression, multiplicity of infection strategy, and scale-up of the process. Study of the differences in the metabolic requirements of infected and non-infected cells is necessary for high cell density processes. In the bioreactor, the specific oxygen uptake rate (OURsp) plays a central role in process scale-up, leading to the specification of the bioreactor operational parameters. Shear stress can also be an important variable for bioreactor operation due to its influence on cell growth and product expression. The determination of the critical variables in process development is discussed, showing the relevance of the mathematical models that have been developed for the insect cells/baculovirus system in process implementation and control.

Animals↗
PubMed1994

Analysis of C-terminally truncated dengue 2 and dengue 3 virus envelope glycoproteins: processing in insect cells and immunogenic properties in mice.

We constructed two recombinant Autographa californica nuclear polyhedrosis baculoviruses. Spodoptera frugiperda (Sf9) cells containing these constructs produce carboxy-terminally truncated envelope E proteins representing dengue (DEN) virus serotypes 2 and 3. The two recombinant proteins contained their homologous signal sequences at the N terminus and were truncated by 71 and 74 amino acids at the C terminus, respectively. This allowed the translocation of the recombinant proteins to the endoplasmic reticulum followed by glycosylation processing and secretion into the extracellular medium. An additional unglycosylated form which was not secreted was detected inside the infected Sf9 cells. Sera from Swiss mice immunized with the infected Sf9 cell lysates gave a DEN cross-reactive response in ELISA and substantial amounts of neutralizing antibodies to the homologous virus. Similar antibody titres were obtained when the two recombinant proteins were inoculated concomitantly. BALB/c mice were vaccinated with three doses of the recombinant E proteins, taken as monovalent or bivalent immunogens, and challenged with mouse-adapted DEN-2 virus. DEN-2 E protein induced a good protection (90%) against lethal encephalitis and recombinant DEN-3 E protein gave a substantial cross-protection (54%). Eighty-two percent of the mice immunized with a mixture of both recombinant E proteins survived the DEN-2 virus challenge.

Animals↗
PubMed1967

Electron microscopy of a plant-pathogenic virus in the nervous system of its insect vector.

The central nervous system of wound tumor virus (WTV)-infected Agallia constricta was studied by electron microscopy to obtain information concerning the virus distribution in the nervous system. Wound tumor virions were mostly found in the cytoplasm of the ganglion cells and less frequently in the glial cells. WTV was occasionally observed in the perineurium cells, nerve axons, tracheoblasts, and lateral nerves. In the ganglion cells, virions appeared as individual isolated particles (V(1)), in tubular formation (V(2)), and occasionally in aggregates (V(3)). In the glial cells, the virions were mostly seen in the V(3) formation, and very seldom in the V(1) and V(2) formations. In the perineurium cells and tracheoblasts, only small V(3) formations were observed. The isolated virions were usually surrounded with polyribosomes, and often appeared around the foci of the viroplasm. Sometimes degenerating ganglion cells infected with the WTV were encountered. These damaged cells strongly indicated that WTV exerted a cytopathogenic effect on the nerve cells.

Central Nervous System↗