NGF and anti-transferrin receptor antibody conjugate: short and long-term effects on survival of cholinergic neurons in intraocular septal transplants.
We describe a new molecular carrier system that allows for the transport of nerve growth factor (NGF) across the blood-brain barrier (BBB), as assessed by trophic effects on intraocular forebrain transplants that contain central cholinergic neurons. The carrier system involves monoclonal antibodies (OX-26) directed against the transferrin receptor, to which NGF molecules are covalently linked. Transferrin receptors are highly concentrated on brain blood vessels and participate in the transport of iron across the BBB. Host rats with septal transplants were divided into four groups, which received OX-26-NGF, OX-26, NGF or saline intravenously at 2, 4, 6 and 8 weeks after grafting. Half of the animals were killed directly after the final injection, whereas the other half were allowed to survive for an additional 5 months. Control experiments revealed that blood vessels in mature brain grafts in oculo contained large amounts of transferrin receptors. Covalent binding of NGF to the OX-26 antibodies did not impede OX-26 binding to CNS transferrin receptors, nor did conjugation affect the bioactivity of NGF. A time-dependent increase in host brain NGF levels was found after injection of OX-26-NGF into the tail vein. Host serum contained some NGF antibodies in the short-term OX-26-NGF group that had disappeared in the long-term group; host adrenals showed no differences in wet weight or norepinephrine or epinephrine whole tissue levels in any of the groups. As previously reported, the overall growth of intraocular septal transplants was approximately twice as great in the OX-26-NGF group relative to all other groups. This difference in final size persisted unabated for at least 5 months after the last injection. Furthermore, the significantly higher numbers of choline acetyl transferase immunoreactive neurons in transplants of OX-26-NGF-treated hosts also persisted during the 5-month postinjection interval. Taken together, the data suggest that the OX-26 conjugate may be a unique approach to permit passage of neurotrophin peptides into the brain in a biologically active form.
Granholm
A C
AC
Department of Basic Science, University of Colorado School of Dentistry, Denver.
Bäckman
C
C
Bloom
F
F
Ebendal
T
T
Gerhardt
G A
GA
Hoffer
B
B
Mackerlova
L
L
Olson
L
L
Söderström
S
S
Walus
L R
LR
D000311:Adrenal Glands / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000917:Antibody Formation; D001812:Blood-Brain Barrier; D001921:Brain / Q000098:blood supply; D016503:Drug Delivery Systems; D004797:Enzyme-Linked Immunosorbent Assay; D004837:Epinephrine / Q000378:metabolism; D005260:Female; D006624:Hippocampus / Q000166:cytology* / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D009414:Nerve Growth Factors / Q000008:administration & dosage / Q000276:immunology / Q000494:pharmacology*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000637:transplantation*; D009638:Norepinephrine / Q000378:metabolism; D013508:Ophthalmologic Surgical Procedures; D051381:Rats; D017207:Rats, Sprague-Dawley; D011950:Receptors, Cholinergic / Q000378:metabolism; D011990:Receptors, Transferrin / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D009414:Nerve Growth Factors; D011950:Receptors, Cholinergic; D011990:Receptors, Transferrin; D009638:Norepinephrine; D004837:Epinephrine