Accessibility to intracellular antigens within nutritionally deprived human mammary epithelial cells.
We have previously demonstrated immunolocalization of antikeratin antibodies in apparently random subpopulations of malignant cells in fresh surgical specimens of breast carcinoma (S. H. Dairkee and A. J. Hackett, 1988, J. Natl. Cancer Inst. 80, 1216-1220). The goal of the present study was to determine whether deficiencies in essential nutrients contribute toward cellular alterations in membrane integrity, consequently allowing antikeratin to bind to the cytoskeleton within live, unfixed cells. We have demonstrated here that in an in vitro model in which human mammary epithelial cells are subjected to an oxygen-glucose gradient, immunolocalization of antikeratin within the cells is observed in a dose-dependent manner in the depleted regions of the gradient, even though the cells appear to be morphologically unaltered. The potential use of antibodies to intracellular antigens for immunotargeting solid tumors and the use of this method in antibody-loading studies toward understanding functional aspects of specific cellular antigens, as well as determining differential response of various cell types under these culture conditions, are discussed.
Dairkee
S H
SH
Peralta Cancer Research Institute, Oakland, California 94609.
Puett
L
L
Counelis
A M
AM
Hackett
A J
AJ
D001386:Azides / Q000494:pharmacology; D001940:Breast / Q000166:cytology; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D003584:Cytological Techniques; D003593:Cytoplasm / Q000737:chemistry; D003599:Cytoskeleton / Q000737:chemistry; D004847:Epithelial Cells; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D007633:Keratins / Q000032:analysis; D010100:Oxygen / Q000378:metabolism; D019810:Sodium Azide
D001386:Azides; D007633:Keratins; D019810:Sodium Azide; D005947:Glucose; D010100:Oxygen