Mukhtar M, Bagasra O, Duan L, Amjad M, Sarker A, Bobroski L, Pomerantz RJ; Conference on Retroviruses and Opportunistic Infections.

Program Abstr 4th Conf Retrovir Oppor Infect Conf Retrovir Oppor Infect 4th 1997 Wash DC. 1997 Jan 22-26; 4th: 89 (abstract no. 136).

Dorrance H. Hamilton Laboratories, Center for Human Virology, Philadelphia, PA.

Recent advances in gene delivery technology offer considerable optimism for ameliorating a number of neurological disease stages including inherited and non-inherited disorders. The pathways and contributing determinants of HIV-1 invasion of the nervous system are relatively unknown. Significant control of viral replication by single chain intracellularly expressed antibodies prompted us to utilize these strategies for controlling HIV-1 in CNS-based cells. We have developed an in vitro Blood Brain Barrier (BBB) system utilizing primary culture of human brain derived microvascular endothelial cells (MVECs), macrophages, neurons and astrocytes. The BBB system developed in our laboratories simulates important morphological and permeability characteristics of the BBB in vivo. In a two compartment system, MVECs were cultured to 100% confluency on the upper side of 0.45 micrometers polycarbonate membrane insert coated with MVECs attachment factor and other CNS cells either alone or in combination in the cover portion of six well chamber inserts. MVECs on the insert were exposed to HIV-1 strains 89.6, NL4-3 and IIIB overnight. After 72 hours the cells were fixed and utilized for in situ PCR. Utilizing transduction of gene therapeutic constructs into MVECs, we studied the expression of an anti-Rev single-chain variable fragment (SFv) in murine retroviral vectors, previously constructed in our laboratories. It was observed that macrophage tropic strain 89.6 has more potential to create a breach in in vitro blood brain barrier, followed by pNL4-3 and IIIB. p24 determination showed that mature neurons were the most susceptible to infection by all three strains. In situ PCR reveals 70-80% infection of the neuronal cells. Transduction of recombinant antibody fragment against Rev significantly inhibited viral replication in MVECs. Our studies reveal an in vitro BBB from human brain cells as well as the use of molecular therapeutic moieties to inhibit HIV-1 replication.

Publication Types:

• Meeting Abstracts

Keywords:

• Astrocytes
• Blood-Brain Barrier
• Brain
• Gene Therapy
• HIV-1
• Humans
• In Vitro
• Macrophages
• Neurons

Other ID:

• 97926272

UI: 102223281

From Meeting Abstracts