title: In vitro effects of anti-HIV immunotoxins directed against multiple epitopes on HIV type 1 envelope glycoprotein 160.
abstract: We have used a panel of anti-gp160 MAbs to construct anti-HIV immunotoxins by coupling antibodies to ricin A chain (RAC). The ability of the immunotoxins to kill HIV-1-infected cells and halt the spread of infection was tested in tissue culture on persistently and acutely infected cell lines and primary lymphocyte cultures stimulated with phytohemagglutinin (PHA blasts). Laboratory strains and clinical isolates of HIV both were tested. The constitution and antigen-binding capacity of the immunotoxins were confirmed by ELISA and indirect immunofluorescence. Immunotoxins that bind epitopes exposed on the cell surface effectively killed persistently infected cells, although killing was not directly proportional to binding of immunotoxin to cell. The activity of anti-gp41, but not anti-gp120, immunotoxins was markedly enhanced in the presence of soluble CD4 or peptides corresponding to the CDR3 region of CD4. CD4-mediated enhancement of anti-gp41 immunotoxin activity was observed for laboratory strains neutralized by sCD4 and for clinical isolates that were resistant to neutralization by sCD4. Immunotoxin action was potentiated by brefeldin A, bafilomycin A1, cortisone, and an amphipathic fusion peptide, but not by cytochalasin D, nocodazol, monodansyl cadaverine, or trans-retinoic acid. Anti-HIV immunotoxins are useful tool with which to study the functional expression of gp120/gp41 antigens on the surface of HIV-infected cells, as well as potential AIDS therapeutics. Because these studies relate to the accessibility of viral antigens to antibody-mediated attack, these studies also have relevance for vaccine development.
authors.affiliation: Laboratory of Microbial Structure and Function, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 59840, USA.
authors.forename: S H
authors.initials: SH
authors.lastname: Pincus
authors.forename: K
authors.initials: K
authors.lastname: Wehrly
authors.forename: R
authors.initials: R
authors.lastname: Cole
authors.forename: H
authors.initials: H
authors.lastname: Fang
authors.forename: G K
authors.initials: GK
authors.lastname: Lewis
authors.forename: J
authors.initials: J
authors.lastname: McClure
authors.forename: A J
authors.initials: AJ
authors.lastname: Conley
authors.forename: B
authors.initials: B
authors.lastname: Wahren
authors.forename: M R
authors.initials: MR
authors.lastname: Posner
authors.forename: A L
authors.initials: AL
authors.lastname: Notkins
authors.forename: S A
authors.initials: SA
authors.lastname: Tilley
authors.forename: A
authors.initials: A
authors.lastname: Pinter
authors.forename: L
authors.initials: L
authors.lastname: Eiden
authors.forename: M
authors.initials: M
authors.lastname: Teintze
authors.forename: D
authors.initials: D
authors.lastname: Dorward
authors.forename: V V
authors.initials: VV
authors.lastname: Tolstikov
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000935:Antifungal Agents / Q000494:pharmacology; D020126:Brefeldin A; D002460:Cell Line; D003348:Cortisone / Q000494:pharmacology; D003517:Cyclopentanes / Q000494:pharmacology; D015638:Cytochalasin D / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D019302:HIV Envelope Protein gp160 / Q000276:immunology*; D015700:HIV Envelope Protein gp41 / Q000276:immunology; D015497:HIV-1 / Q000187:drug effects / Q000276:immunology*; D006801:Humans; D000922:Immunotoxins / Q000494:pharmacology*; D018942:Macrolides; D008855:Microscopy, Electron, Scanning; D015739:Nocodazole / Q000494:pharmacology; D010835:Phytohemagglutinins / Q000276:immunology
chemical_list: D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D003517:Cyclopentanes; D000939:Epitopes; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41; D000922:Immunotoxins; D018942:Macrolides; D010835:Phytohemagglutinins; D020126:Brefeldin A; D015638:Cytochalasin D; C040929:bafilomycin A1; D015739:Nocodazole; D003348:Cortisone