Proteoglycans secreted by packaging cell lines inhibit retrovirus infection.
Using a model recombinant retrovirus encoding the Escherichia coli lacZ gene, we have found that medium conditioned with NIH 3T3 cells and packaging cell lines derived from NIH 3T3 cells inhibits infection. Most of the inhibitory activity was greater than 100 kDa and was sensitive to chondroitinase ABC digestion, which is consistent with the inhibitor being a chondroitin sulfate proteoglycan. Proteoglycans secreted by NIH 3T3 cells and purified by anion-exchange chromatography inhibited amphotropic retrovirus infection. Pretreatment of amphotropic retrovirus stocks with chondroitinase ABC boosted the level of transduction efficiency by more than twofold. The implications of these findings with respect to retrovirus-cell interactions and the production of high-titer retroviral stocks are discussed.
Le Doux
J M
JM
Department of Chemical and Biochemical Engineering, Rutgers University, Piscataway, New Jersey 08854, USA.
Morgan
J R
JR
Snow
R G
RG
Yarmush
M L
ML
D016475:3T3 Cells; D000818:Animals; D011508:Chondroitin Sulfate Proteoglycans / Q000096:biosynthesis / Q000378:metabolism* / Q000494:pharmacology; D002810:Chondroitinases and Chondroitin Lyases; D017077:Culture Media, Conditioned; D003851:Deoxyribonucleases; D010450:Endopeptidases; D005347:Fibroblasts; D005822:Genetic Vectors; D019763:Heparin Lyase; D006358:Hot Temperature; D007700:Kinetics; D007763:Lac Operon; D051379:Mice; D011133:Polysaccharide-Lyases; D011509:Proteoglycans / Q000378:metabolism* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis; D012190:Retroviridae / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D012260:Ribonucleases; D014357:Trypsin; D001616:beta-Galactosidase / Q000096:biosynthesis*
D011508:Chondroitin Sulfate Proteoglycans; D017077:Culture Media, Conditioned; D011509:Proteoglycans; D011994:Recombinant Proteins; D003851:Deoxyribonucleases; D012260:Ribonucleases; D001616:beta-Galactosidase; D010450:Endopeptidases; D014357:Trypsin; D002810:Chondroitinases and Chondroitin Lyases; D011133:Polysaccharide-Lyases; D019763:Heparin Lyase