Retroviral vector producer cell killing in human serum is mediated by natural antibody and complement: strategies for evading the humoral immune response.
The introduction of retroviral vector producer cells (VPC) into tumors as a means of increasing transduction efficiency has recently been employed in human gene therapy trials. However, the fate of these xenogeneic cells in humans is not well understood. In the present study, we used an in vitro model to examine the survival of commonly used VPC lines in serum from humans and various other species. VPC derived from the murine NIH-3T3 cell line, including PA317, Psi CRIP, and GP + E-86, were effectively killed in sera from Old World primates, including human and baboon. Conversely, the same murine cell lines survived exposure to sera from dog, rabbit, rat, and mouse. This pattern of serum killing parallels the occurrence of the anti-alpha-galactosyl natural antibody (Ab) found exclusively in Old World primates. The anti-alpha-galactosyl Ab targets the terminal glycosidic structure Gal alpha 1-3Gal beta 1-4GlcNAc-R (alpha-galactosyl epitope) found on the surface of mammalian cells, excluding Old World primates. All murine-derived VPC tested expressed high levels of the alpha-galactosyl epitope as determined by FACS analysis. VPC killing was complement-mediated, because preincubation of human serum with a functionally blocking anti-C5 mAb completely abolished cell lysis. Furthermore, addition of soluble galactose(alpha 1-3)galactose (Gal alpha 1-3Gal) to human serum or down-regulation of the alpha-galactosyl epitope on the surface of VPC effectively reduced VPC killing, indicating that complement activation by these cells is primarily initiated by natural antibody recognition of the alpha-galactosyl epitope. Finally, VPC incubated with human serum for 8 hr in the presence of complement inhibition continued to produce viable retroviral particles, thus demonstrating a correlation between VPC and particle survival. Taken together, these data suggest that elimination of the alpha-galactosyl epitope or complement blockade may provide a strategy to prolong the survival of VPC and the particles that they produce in vivo.
Rollins
S A
SA
Department of Immunobiology, Alexion Pharmaceuticals Inc., New Haven, CT 06511, USA.
Birks
C W
CW
Setter
E
E
Squinto
S P
SP
Rother
R P
RP
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D002240:Carbohydrate Sequence; D002470:Cell Survival / Q000235:genetics*; D002478:Cells, Cultured; D003165:Complement System Proteins / Q000276:immunology*; D004285:Dogs; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D005690:Galactose / Q000276:immunology*; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009374:Neoplasms, Experimental / Q000235:genetics / Q000276:immunology / Q000628:therapy; D010215:Papio / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D012190:Retroviridae / Q000235:genetics* / Q000378:metabolism
D000906:Antibodies; D000939:Epitopes; D003165:Complement System Proteins; D005690:Galactose