Human blood basophils synthesize interleukin-2 binding sites.
Recent data suggest that basophils express receptors for a variety of lymphokines. In this study we present the biochemical characterization of the interleukin-2 (IL-2) receptor on the basophil surface membrane. Highly enriched populations (purity: 92% to 99%) of blood basophils were obtained from chronic granulocytic leukemia (CGL) patients (n = 3) by negative selection using monoclonal antibodies (MoAbs) and complement. CGL basophils were found to bind CD25 MoAbs (n = 4) directed against different epitopes of the 55- to 60-Kd subunit of the IL-2 receptor (= Tac peptide). Immunoprecipitation experiments with lysates of purified CGL basophils and CD25 MoAbs showed a protein with a molecular weight of 60 Kd, equivalent to the Tac peptide on human T blasts. Quantitative binding studies and Scatchard plot analysis using radiolabeled recombinant human (rh) IL-2 indicated the presence of 12,000 +/- 4,700 low affinity IL-2 binding sites (kd = 66 nmol/L) per purified CGL basophil. Northern blot analysis with enriched CGL basophils showed two messenger RNA bands of 3.5 and 1.5 kilobases hybridizing to radiolabeled Tac cDNA. Immunoprecipitation of the Tac peptide from enriched basophils metabolically labeled with 35S-methionine showed active synthesis of the IL-2 receptor. Our results show that human blood basophils synthesize and express receptors for IL-2.
Institute of Immunology, University of Vienna, Austria.
H
H
Stockinger
P
P
Valent
O
O
Majdic
P
P
Bettelheim
W
W
Knapp
D000911:Antibodies, Monoclonal; D001491:Basophils / Q000276:immunology* / Q000473:pathology / Q000502:physiology; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D006636:Histamine Release; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000097:blood / Q000276:immunology*; D008970:Molecular Weight; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D015375:Receptors, Interleukin-2