Targeting of phototoxic drugs to antigen-specific T lymphocytes in vitro using antigen-presenting cell membranes.
We have used the complex of antigen with class II major histocompatibility proteins (Ia) in membrane-bound form to target a phototoxic compound to antigen-specific T cell hybridomas in vitro. The iodoacetamidyl ester of phototoxic pyrene was bound covalently to antigen-presenting cells (APC), and protein antigens were added to the cells for processing, presentation and targeting of the drug to three different T hybridomas specific for myelin basic protein (MBP), ovalbumin (OVA) and keyhole limpet hemocyanin (KLH). The B hybridoma LS102.9 was used as APC to present MBP, KLH and either a tryptic digest of OVA or the synthetic peptide OVA323-339 to these T cells. A transformed B lymphoma, which expresses trinitrophenol (TNP)-specific surface IgM, A20-HL, was used to present TNP conjugates of KLH and OVA to T cells. Either the antigen-bearing intact APC or Ia+ membranes shed spontaneously from them were used as drug carriers to target pyrene to the T cells. In the dark, or in the absence of pyrene, both the intact APC or the shed membranes stimulated interleukin-2 (IL-2) production by the T cells in an antigen-specific way. After UVA (320-400 nm) irradiation, both forms of these drug carriers had an antigen-specific toxic effect on the T hybridoma cells with receptors for the antigen that they carried. Both spontaneous T cell proliferation and antigen-induced IL-2 production were inhibited. The shed membranes had a more antigen-specific toxic effect than the intact APC, which tend to settle out with the T cells in the microtiter plates, possibly causing nonspecific contact.(ABSTRACT TRUNCATED AT 250 WORDS)
Chang
N H
NH
Department of Clinical Biochemistry and Immunology, University of Toronto, Canada.
Rayner
D C
DC
Boggs
J M
JM
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D004337:Drug Carriers; D006433:Hemocyanins / Q000008:administration & dosage / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism*; D007376:Interleukin-2 / Q000096:biosynthesis; D016393:Lymphoma, B-Cell; D004676:Myelin Basic Protein / Q000008:administration & dosage / Q000276:immunology; D010047:Ovalbumin / Q000008:administration & dosage / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D004337:Drug Carriers; D000949:Histocompatibility Antigens Class II; D007376:Interleukin-2; D004676:Myelin Basic Protein; D010047:Ovalbumin; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin