Nuclear localizing anti-DNA antibodies enter cells via caveoli and modulate expression of caveolin and p53.
After administration to normal mice, a subset of monoclonal (m) anti-DNA antibodies (Ab) derived from MRL-lpr/lpr mice was identified that enter cells, in vivo. In the kidneys, this was associated with glomerular hypercellularity and proteinuria. In cultured cells, the same mAb bound to myosin 1 on the cell surface, prior to internalization, nuclear localization and inhibition of apoptosis. The present study focuses on the mechanisms underlying the observed functional effects. Subcellular localization studies revealed that following internalization, a prototypic, nuclear localizing, m antibody (Ab; termed H7) co-localized with myosin 1, shortly after internalization, within caveolae, near the cell membrane. Cell fractionation studies confirmed the presence of both H7 and myosin within the caveolar fraction. Since variations in caveolin protein expression have been associated with apoptotic events in cancer cells, through p53 dependent and independent pathways, modulation of caveolin by intracellular H7 was evaluated. Cellular entry of the anti-DNA Ab resulted in an increase in caveolin protein expression. Furthermore, after exposure of cells to dexamethasone to induce apoptosis, the usual increase in p53 was inhibited in the presence of intracellular H7. Taken together, the results suggest that upregulation of caveolin and inhibition of p53 induction are involved in H7-induced, inhibition of apoptosis. Furthermore, they suggest that this inhibition contributes to the glomerular hypercellularity observed in normal mice with intranuclear H7. The results also raise the possibility that inhibition of apoptotic pathways during inflammation or/and autoimmunity could influence subsequent disease events. The novel mechanism of cellular perturbation is indirect and dependent on apoptotic stimuli, and it may account for the presence of intranuclear antibodies in inflammatory and normal tissues of individuals with lupus.
Yanase
Kumiko
K
Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104-6144, USA.
Madaio
Michael P
MP
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D000974:Antibodies, Antinuclear / Q000276:immunology / Q000378:metabolism*; D021941:Caveolae / Q000276:immunology / Q000378:metabolism*; D051242:Caveolin 1; D022461:Caveolins / Q000378:metabolism*; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D002875:Chromosomes / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D003907:Dexamethasone / Q000037:antagonists & inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation; D016253:Microscopy, Immunoelectron; D009218:Myosins / Q000378:metabolism; D022022:Nuclear Pore / Q000276:immunology / Q000378:metabolism; D051381:Rats; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects
D000974:Antibodies, Antinuclear; C495537:Cav1 protein, rat; D051242:Caveolin 1; D022461:Caveolins; D016159:Tumor Suppressor Protein p53; D003907:Dexamethasone; D000255:Adenosine Triphosphate; D009218:Myosins