B lymphocytes respond specifically to phytohaemagglutinin after liposome-dependent transfer of purified phytohaemagglutinin receptors.
Phytohaemagglutinin (PHA) receptor glycoproteins purified by affinity chromatography from porcine splenic lymphocytes, were reconstituted into vesicles made of phosphatidylcholine and phosphatidylserine, concomitantly with Sendai virus fusogenic proteins HN and F. The vesicles were used as a vehicle to insert the PHA receptor glycoproteins into a highly enriched population of porcine B lymphocytes. Fluorescence analyses showed that 52 +/- 2% of the reconstituted B cells had incorporated the lectin receptors. The modified B lymphocytes were assayed for their response (tritiated thymidine incorporation into nucleic acids) to PHA, concanavalin A (Con A), or to lipopolysaccharide (LPS). The results showed that porcine B cells fused with vesicles containing only viral fusogenic proteins failed to respond to either PHA or Con A. Tritiated thymidine incorporation was similar to background values. The cells did, however, respond to LPS with values of label incorporation similar to those observed in the case of pre-fused B lymphocytes. When purified B lymphocytes were fused with vesicles containing PHA receptors and viral fusogenic proteins, assays of thymidine incorporation showed a statistically significant (P less than 0.001) and specific response of the modified cells to PHA stimulation. Reconstituted cells cultured in the presence of PHA incorporated approximately nine times more radioactive label than pre-fused cells or cells fused with vesicles containing only fusogenic viral proteins. In marked contrast, reconstituted B lymphocytes did not show any significant label incorporation above background level in response to Con A, but they retained their ability to respond to LPS. Our findings suggest that B lymphocytes can be made to respond specifically to PHA by insertion of appropriate lymphocyte-derived receptors.
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D002469:Cell Separation; D006023:Glycoproteins / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology; D008081:Liposomes; D008213:Lymphocyte Activation; D008561:Membrane Fusion; D008565:Membrane Proteins / Q000276:immunology; D011975:Receptors, Mitogen / Q000276:immunology* / Q000302:isolation & purification; D013552:Swine
D006023:Glycoproteins; D008070:Lipopolysaccharides; D008081:Liposomes; D008565:Membrane Proteins; D011975:Receptors, Mitogen