The investigators propose to study inhibition of HIV replication in vitro and in vivo by single chain Fv antibodies (SFv) against HIV proteins. They have found that SFv (#Aw) against HIV-1 integrase (IN) inhibits virus replication better than SFv vs. the other HIV proteins we have tested. To deliver SFv genes, they will use a unique gene transfer system that we have developed: T-antigen-depleted SV40 effectively delivers transgene expression in vitro and in vivo to bone marrow cells and to unselected, resting human lymphocytes. SV-Aw delivers very high levels of the HIV-inhibitory SFv, Aw, to cultured human T cells. This transduction system will be used to inhibit HIV-1, SHIV and SIV in vitro and in vivo. Their group of collaborators will apply test systems that they have developed: a gene transfer vector that yields high level transgene expression on inoculation in vivo, intracellular SFv that inhibit HIV-1 in vitro, a mouse system to study progression and inhibition of HIV infection in bone marrow-derived cells, and a primate system to test SHIV and SIV infection and its inhibition. The applicants hypothesize that intracellular anti-IN SFv can be transferred by SV-40-derived vectors sufficiently to protect against HIV and SIV infection in vitro and in vivo. To test this hypothesis, they will assess the ability of SV40-delivered SFv to inhibit HIV infection of human T lymphocytes in vitro by testing SV-Aw, as well as SV40-delivered SFv vs. HIV rev and reverse transcriptase. They will apply these vectors to study inhibition of HIV infection by transduction ex vivo, and in vivo, in SCID-hu mice. These studies will identify aspects of SV40-delivered Aw expression that need to be improved, e.g., increasing levels, stability and/or longevity of transgene expression. The applicants will modify transfer vectors accordingly. These enhanced vectors will be tested first in vitro then in vivo vs. HIV-1 in human cells and SCID-hu mice. Because of the importance of studying this approach to HIV inhibition in an experimental system resembling human AIDS, they will use rhesus macaque monkeys to study the ability of SV-40-delivered SFv to inhibit immunosuppressive lentivirus infection in primates. Existing anti-HIV rev SFv will be used to study inhibition of SHIV-4 infection in monkey bone marrow cells, and , if successful, in vivo. They are also currently developing SFv vs. SIV IN, RT and rev for use against SIV in vitro and in vivo. SV40-based gene transduction has great promise, particularly when combined with intracellular SFv, in inhibition of HIV infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI041399-03
Application #
2887456
Study Section
AIDS and Related Research Study Section 1 (ARRA)
Program Officer
Sarver, Nava
Project Start
1997-09-15
Project End
2001-07-31
Budget Start
1999-09-01
Budget End
2001-07-31
Support Year
3
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Thomas Jefferson University
Department
Pathology
Type
Schools of Medicine
DUNS #
061197161
City
Philadelphia
State
PA
Country
United States
Zip Code
19107
Rose, Sasha J; Bermudez, Luiz E (2016) Identification of Bicarbonate as a Trigger and Genes Involved with Extracellular DNA Export in Mycobacterial Biofilms. MBio 7:
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