Implanted IgE-Fc epsilon R complexes elicit IgE-mediated activation of RBL-2H cells.
The high-affinity receptor for IgE (Fc epsilon R) is the cellular trigger of the antigen-induced activation of mast cells and basophils. To examine the functional integrity of Fc epsilon R, we have adopted a protein implantation procedure whereby the purified receptor complex was coreconstituted with Sendai virus envelopes. The latter promoted fusion of the hybrid vesicles with recipient cells such as rat basophilic leukemia, RBL-2H3, thus serving as a vehicle for the receptor. The implanted Fc epsilon R was complexed with 125I-labeled mouse IgE (anti-DNP) to permit receptor quantification as well as specific triggering by DNP20BSA. Implantation in the presence of unlabeled rat IgE, which blocked the native receptors on the recipient RBL-2H3 cells, resulted in incorporation of up to 15 ng of receptor-bound IgE/10(6) cells. This was roughly equivalent in amount to 10-20% of the native receptors on such cells. The exocytosis which was triggered in the recipient cells by reagents that specifically recognized the implanted IgE reached between 15 and 50% of the maximal response. Various treatments that interfered with the activities of the viral envelopes reduced both receptor incorporation (3-5-fold) and cell degranulation (3-10-fold). These included separation of the receptor from the reconstituted envelopes, addition of serum to the incubation mixture (to inhibit vesicle-cell binding), and trypsinization of the virus (to inhibit vesicle-cell fusion). Poly(ethylene glycol) 8000 (4%) enhanced both the incorporation of the receptor and its functional responses. These treatments distinguished between real incorporation of IgE-Fc epsilon R complexes and other mechanisms of 125I-IgE association with the recipient cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Ran
S
S
Department of Membrane Research, Weizmann Institute of Science, Rehovot, Israel.
Loyter
A
A
Rivnay
B
B
D000818:Animals; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D014644:Genetic Variation; D007073:Immunoglobulin E / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D015471:Leukemia, Basophilic, Acute / Q000276:immunology* / Q000378:metabolism; D010222:Parainfluenza Virus 1, Human; D051381:Rats; D011961:Receptors, Fc / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D017455:Receptors, IgE; D014759:Viral Envelope Proteins / Q000378:metabolism
D000944:Antigens, Differentiation, B-Lymphocyte; D011961:Receptors, Fc; D017455:Receptors, IgE; D014759:Viral Envelope Proteins; D007073:Immunoglobulin E; D002118:Calcium