Identification of the interleukin-2 receptor (IL-2R) on human leukemic T cells using colloidal gold and scanning electron microscopy.
Results of studies demonstrating the identification of the interleukin-2 receptor (IL-2R; i.e., anti-Tac) on the membrane ultra-structure of human leukemic T cells with an antibody carrying an electron dense colloidal gold microsphere (e.g., immunogold) that was visualized using a scanning electron microscope (SEM) are reported. Our IL-2R model system employed HTLV-1 retrovirus-infected lymphoblastoid cells of the long-term human leukemic T cell line HUT-102B2. The presence of the IL-2R on these cells was defined using a double antibody procedure that employed as the primary antibody a purified mouse monoclonal anti-Leu-IL-2R antibody (mIgGlk, anti-Tac, CD25), and used as the secondary antibody a goat anti-mouse IgG (gamma-chain specific) antibody that had been covalently bonded to a 40 nm colloidal gold particle. More than 95% of the HUT-102B2 were IL-2R+, and there was a uniform distribution of the IL-2R over the surface of the cells. Corresponding controls were employed in all examinations and included IL-2R- Jurkat human leukemic T cells and isotype identical immunoglobulins. The primary and secondary antibody reagents contained whole human serum and bovine serum albumin, and there was no evidence of the non-specific binding of these antibodies. These studies are the first to demonstrate the presence of a lymphokine receptor on the surface architecture of a cell. We anticipate no difficulty in applying the immunogold/SEM technology to define both normal and malignant cell membrane receptors for other cytokines.
Helinski
E H
EH
Department of Molecular Immunology, Roswell Park Memorial Institute, Buffalo, NY 14263.
Bielat
K L
KL
Ovak
G M
GM
Meenaghan
M A
MA
Wirth
J E
JE
Pauly
J L
JL
D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure; D003102:Colloids; D005434:Flow Cytometry; D006046:Gold; D006801:Humans; D007938:Leukemia / Q000473:pathology*; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D015375:Receptors, Interleukin-2 / Q000648:ultrastructure*; D013601:T-Lymphocytes / Q000648:ultrastructure*
D003102:Colloids; D015375:Receptors, Interleukin-2; D006046:Gold