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Arthritis Center, Department of Medicine, Boston University School of Medicine, Boston, MA 02118
Culture supernatants from retroviral packaging cells carrying the
human Fas ligand (FasL) gene killed both human (Jurkat) and mouse
(LB27.4) targets within 5 h of incubation. Cytotoxicity was found
both in a fraction 
500 kDa and a fraction between 50 and 500 kDa.
Following ultracentrifugation, the activity in the 500-kDa fraction
was concentrated in the pellet (FasL vector preparation (VP)), which
was also infective when added to NIH-3T3 cells. Both Polybrene and
poly-L-lysine significantly enhanced the cytotoxicity of
FasL VP but not anti-Fas mAb, soluble FasL (sFasL), and
cell-associated FasL. In the presence of Polybrene, FasL VP killed
targets that are resistant to anti-Fas mAb and sFasL. The
infectivity but not FasL cytotoxicity of FasL VP was sensitive to
irradiation and heat shock. By contrast, cytotoxicity of FasL VP could
be enhanced or inhibited depending on the doses of anti-FasL mAb.
Interestingly, the infectivity of FasL VP was specifically enhanced by
anti-FasL mAb, suggesting that a nonviral gene product could be
used to regulate the behavior of the retroviral vector. Thus, in
addition to expressing potent FasL cytotoxicity, the FasL VP exhibits
unique properties heretofore not attributed to anti-Fas mAb, sFasL,
and cell-associated FasL. Our study raises the possibility of using the
retroviral gene-packaging technology to make powerful, versatile, and
regulatable bioactive vesicles expressing a predetermined function of
the protein encoded by the target gene.
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