Localization of interferon-induced lupus inclusions demonstrated by computer image reconstruction of monensin-treated Daudi cells.
A structural analysis of cells that contained the interferon-alpha-induced lupus inclusions (LI) was performed using a high-voltage electron microscope to determine the exact cellular location of LI and their association with normal cell organelles. LI were induced in the human B lymphoblastoid cell line, Daudi, by culturing with the pure recombinant human leukocyte interferon, IFLrA. Just prior to harvesting, a portion of the cells was treated with monensin to selectively swell the Golgi apparatus, and thereby simplify their identification using the electron microscope. Organellar associations between LI and the outer nuclear envelope and Golgi apparatus were identified in stereopairs of 1-micron sections prepared from both cells that were not treated with monensin and those that were treated with monensin. Serial 0.25-micron sections of the monensin-treated cells were prepared, and seven arbitrarily chosen cells were examined. Each of these cells contained a single LI, and it formed throughout an endoplasmic-reticulum region that made contact with both the outer nuclear envelope and the Golgi vesicles. Reconstruction of a cell by computer from the digitized negatives of serial sections clearly illustrated these relationships. This study reports the first determination of the association between LI and the Golgi apparatus. It also identifies the presence of only one LI in every cell, and the routine association of the LI with both the outer nuclear envelope and the Golgi apparatus. The unique cell location of LI formation suggests their functioning in membrane biogenesis, the trafficking of proteins to the plasma membrane or to cytoplasmic vesicles, or the processing of proteins for secretion.
Laboratory of Biophysics, Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
S A
SA
Rich
M
M
Marko
W E
WE
Gibbons
D000163:Acquired Immunodeficiency Syndrome / Q000473:pathology; D001402:B-Lymphocytes / Q000187:drug effects / Q000648:ultrastructure; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000187:drug effects / Q000648:ultrastructure; D006801:Humans; D007091:Image Processing, Computer-Assisted; D002479:Inclusion Bodies / Q000648:ultrastructure*; D007372:Interferons / Q000502:physiology*; D008177:Lupus Vulgaris / Q000473:pathology; D008854:Microscopy, Electron; D008985:Monensin / Q000494:pharmacology; D012157:Mononuclear Phagocyte System / Q000648:ultrastructure*; D009685:Nuclear Envelope / Q000648:ultrastructure; D016511:Severe Combined Immunodeficiency / Q000473:pathology; D014407:Tumor Cells, Cultured
D007372:Interferons; D008985:Monensin