The importance of the acquired immunodeficiency syndrome (AIDS) needs no discussion. The present proposal will create a comprehensive national cooperative drug discovery group based upon novel thematic approaches to understand and control this disease. Five sets of investigators will collectively exploit their unique skills in immunology, chemistry, cell biology and molecular biology in a dynamic and interdigitating fashion. Employing recombinant technology in a baculoviral system, the Reinherz group has produced a secreted form of the CD4 extracellular segment which avidly binds the HIV gp120 without affecting normal immune responses directed against MHC class II. Synthetic CD4 peptides, truncated CD4 proteins and CD4 proteins altered by site-directed mutagenesis as wellas side chain modifications will lead to elucidation of the gp120 binding region. Crystallographic resolution of CD4 will be undertaken collaboratively with Drs. Wiley and Harrison. At the same time, will exploit their skills in immunotoxin (IT) chemistry and the unique CD4-gp120 interaction in order to develop CD4-toxins for targeting the death of HIV-infected cells. Several immunochemical as well as recombinant approaches with various CD4 analogs will lead to discovery of immunotoxins with optimal pharmacokinetics for in vivo use. We are systemically identifying peptide epitopes on gp120 which are recognized by CTL from healthy, uninfected individuals, thus providing important information for vaccine design as well as for selective epitope targeting by immunologic effector cells. We will combine their expertise in enzymology and gene expression to characterize existing HIV POL over-expressors they have produced in the baculovirus system. Analysis of both forms of reverse trancriptase (RT), integrase, RNAase H and protease in this eukaryotic system will provide critical substrates for drug design and screening. The potential for delivering these compounds to HIV-infected cells via the CD4 structure will be investigated.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01AI027336-04
Application #
3547238
Study Section
Special Emphasis Panel (SRC (34))
Project Start
1988-09-30
Project End
1993-08-31
Budget Start
1991-09-01
Budget End
1992-08-31
Support Year
4
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Dana-Farber Cancer Institute
Department
Type
DUNS #
149617367
City
Boston
State
MA
Country
United States
Zip Code
02215
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Tan, K; Liu, J; Wang, J et al. (1997) Atomic structure of a thermostable subdomain of HIV-1 gp41. Proc Natl Acad Sci U S A 94:12303-8
Ferrer, M; Godbout, K L; Sullivan, B J et al. (1997) Construction and characterization of a radio-iodinatable mutant of recombinant human CD4. J Immunol Methods 210:215-25
Sakihama, T; Smolyar, A; Reinherz, E L (1995) Oligomerization of CD4 is required for stable binding to class II major histocompatibility complex proteins but not for interaction with human immunodeficiency virus gp120. Proc Natl Acad Sci U S A 92:6444-8
Sakihama, T; Smolyar, A; Reinherz, E L (1995) Molecular recognition of antigen involves lattice formation between CD4, MHC class II and TCR molecules. Immunol Today 16:581-7
Bell, K D; Ramilo, O; Vitetta, E S (1993) Combined use of an immunotoxin and cyclosporine to prevent both activated and quiescent peripheral blood T cells from producing type 1 human immunodeficiency virus. Proc Natl Acad Sci U S A 90:1411-5
Moebius, U; Pallai, P; Harrison, S C et al. (1993) Delineation of an extended surface contact area on human CD4 involved in class II major histocompatibility complex binding. Proc Natl Acad Sci U S A 90:8259-63
Moebius, U; Clayton, L K; Abraham, S et al. (1992) The human immunodeficiency virus gp120 binding site on CD4: delineation by quantitative equilibrium and kinetic binding studies of mutants in conjunction with a high-resolution CD4 atomic structure. J Exp Med 176:507-17
Moebius, U; Clayton, L K; Abraham, S et al. (1992) Human immunodeficiency virus gp120 binding C'C"" ridge of CD4 domain 1 is also involved in interaction with class II major histocompatibility complex molecules. Proc Natl Acad Sci U S A 89:12008-12
Ghetie, V; Wheeler, T; Scott, D et al. (1992) A CD4-derived peptide carrier blocks acute HIV-1 infection in vitro and binds to gp120 in the presence of Walter-Reed stage 1-6 HIV+ sera. AIDS Res Hum Retroviruses 8:1945-8

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