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ARS Home » Research » Publications at this Location » Publication #148301
Title: TRYPSINIZATION OF OCCLUSION BODY-DERIVED VIRUS FROM THREE NUCLEOPOLYHEDROVIRUSES ALTERS INFECTIVITY OF THREE VIRUS ISOLATES TO INSECT CELL LINES.

Author
item LYNN, DWIGHT

Submitted to: Society for Invertebrate Pathology Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 5/26/2003
Publication Date: 7/20/2003
Citation: Lynn, D.E. 2003. Trypsinization of occlusion body-derived virus from three nucleopolyhedroviruses alters infectivity of three virus isolates to insect cell lines.. Society for Invertebrate Pathology Annual Meeting.

Interpretive Summary:

Technical Abstract: Twelve insect cell lines were tested for susceptibility to baculovirus infection by use of a typical endpoint assay procedure using alkali-liberated occlusion body-derived virus (ODV). Cell lines from Spodoptera frugiperda (IPLB-Sf21AE), Anticarsia gemmatalis (UFL-Ag286), Heliothis virescens (IPLB-HvE1a, IPLB-HvE6a, IPLB-HvE6s and IPLB-HvT1), Lymantria dispar (IPLB-LdEIta and IPLB-LdEp), Plutella xylostella (IPLB-PxE2), and Trichoplusia ni (TN-368, IAL-TND1, and IPLB-TN-R2) in 96-well tissue culture plates were each infected with dilutions of ODV from three nucleopolyhedroviruses (NPVs), including Autographa californica NPV (AcMNPV), Anagrapha falcifera NPV (AfMNPV), and A. gemmatalis NPV (AgMNPV). Additionally, the ODV of each virus was treated with trypsin (0.05 mg/ml) prior to inoculation of cells (=ODV-T). The resulting virus titers reveal the relative infectivity of ODV and ODV-T from the three viruses to each cell line. Trypsin causes a slight (2- to 5-fold), but consistent, increase in infectivity for each virus in many of the lines tested. Of particular interest, however are the results with AcMNPV and AfMNPV on the LdEp and TN-368 lines. The titers in LdEp are always slightly lower with trypsin than without, although this line also has the highest level of susceptibility to ODV from AcMNPV and AfMNPV and is second only to Ag286 with AgMNPV ODV-T. Alternatively, AcMNPV and AfMNPV ODV show a very large (over 5,000-fold) increase in infectivity to the TN-368 line after trypsinization. Since proteases are a natural feature in the insect midgut where ODV typically initiates the infection of the insect, these results with trypsin might be expected. However, the mode-of-action is currently unclear.