Soluble forms of the cell adhesion molecule L1 produced by insect and baculovirus-transduced mammalian cells enhance Schwann cell motility.
For biotechnological applications, insect cell lines are primarily known as hosts for the baculovirus expression system that is capable to direct synthesis of high levels of recombinant proteins through use of powerful viral promoters. Here, we demonstrate the implementation of two alternative approaches based on the baculovirus system for production of a mammalian recombinant glycoprotein, comprising the extracellular part of the cell adhesion molecule L1, with potential important therapeutic applications in nervous system repair. In the first approach, the extracellular part of L1 bearing a myc tag is produced in permanently transformed insect cell lines and purified by affinity chromatography. In the second approach, recombinant baculoviruses that express L1-Fc chimeric protein, derived from fusion of the extracellular part of L1 with the Fc part of human IgG1, under the control of a mammalian promoter are used to infect mammalian HEK293 and primary Schwann cells. Both the extracellular part of L1 bearing a myc tag accumulating in the supernatants of insect cultures as well as L1-Fc secreted by transduced HEK293 or Schwann cells are capable of increasing the motility of Schwann cells with similar efficiency in a gap bridging bioassay. In addition, baculovirus-transduced Schwann cells show enhanced motility when grafted on organotypic cultures of neonatal brain slices while they retain their ability to myelinate CNS axons. This proof-of-concept that the migratory properties of myelin-forming cells can be modulated by recombinant protein produced in insect culture as well as by means of baculovirus-mediated adhesion molecule expression in mammalian cells may have beneficial applications in the field of CNS therapies.
Lavdas
Alexandros A
AA
Laboratory of Cellular and Molecular Neurobiology, Hellenic Pasteur Institute, Athens, Greece.
Efrose
Rodica
R
Douris
Vassilis
V
Gaitanou
Maria
M
Papastefanaki
Florentia
F
Swevers
Luc
L
Thomaidou
Dimitra
D
Iatrou
Kostas
K
Matsas
Rebecca
R
D000818:Animals; D000831:Animals, Newborn; D016367:Baculoviridae / Q000378:metabolism*; D051929:CD146 Antigen / Q000737:chemistry / Q000378:metabolism; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D002846:Chromatography, Affinity / Q000379:methods; D018920:Coculture Techniques / Q000379:methods; D005786:Gene Expression Regulation / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology; D006801:Humans; D007313:Insecta; D051379:Mice; D009186:Myelin Sheath / Q000378:metabolism; D039842:Neural Cell Adhesion Molecule L1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016548:Prosencephalon / Q000166:cytology / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D012583:Schwann Cells / Q000502:physiology*; D012584:Sciatic Nerve / Q000166:cytology; D014161:Transduction, Genetic / Q000379:methods
D051929:CD146 Antigen; C497902:MCAM protein, human; D039842:Neural Cell Adhesion Molecule L1