Capping and internalization of a monoclonal antibody-surface antigen complex: a possible mode of interaction of monoclonal antibodies and tumor cells.
Our results demonstrate that upon incubation of 125I-3G5 (a monoclonal IgM against a membrane ganglioside antigen on RINm5F cells) with rat insulinoma RINm5F cell monolayers at 37 degrees C, the IgM is rapidly internalized. Cell-bound radioactivity, detectable within 10 to 15 minutes, reaches a peak at 4 hours. By 24 hours the intracellular radioactivity has decreased to about 37.5% of the 4-hour value, accompanied by an increase in free 125I in the incubation medium. The incubation of 125I-3G5 with RINm5F cell monolayers at 4 degrees C shows that this series of events is inhibited by low temperature. Microautoradiography confirms these events indicating the presence of radiolabeled antibody on the plasma membrane as well as distinct capping processes and diffuse radioactive deposits within the cells as early as 5 to 10 minutes after initiating incubation at 37 degrees C. Electron microscopy autoradiography provides a detailed demonstration of the capping phenomenon and of endocytic vacuoles, followed at later times by the distribution of radioactive deposits throughout the cell. This model constituted by the capping of the 125I-3G5-ganglioside complex on rat insulinoma RINm5F cells may be useful in elucidating a possible mode of interaction of monoclonal antibodies and tumor cells.
Mariani
G
G
Department of Radiology Experimental Nuclear Medicine, New England Deaconess Hospital, Boston, Massachusetts.
Ito
S
S
Nayak
R C
RC
Baranowska-Kortylewicz
J
J
Venkateshan
C N
CN
Van den Abbeele
A D
AD
Eisenbarth
G S
GS
Adelstein
S J
SJ
Kassis
A I
AI
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology*; D001666:Binding Sites, Antibody; D005732:Gangliosides / Q000276:immunology*; D007152:Immunologic Capping; D007340:Insulinoma / Q000276:immunology*; D007457:Iodine Radioisotopes; D051379:Mice; D051381:Rats; D014407:Tumor Cells, Cultured / Q000276:immunology
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D005732:Gangliosides; D007457:Iodine Radioisotopes