Identification of a basophil leukocyte interleukin-3-regulated protein that is identical to IgE-dependent histamine-releasing factor.
This study aimed to identify basophil leukocyte proteins associated with interleukin (IL)-3 and/or anti-IgE activation by two-dimensional (2-D) gel electrophoresis. We noticed one particular protein showing increased synthesis after recombinant human (rh)IL-3 and, to a lesser extent, anti-IgE stimulation. The protein was also present in the culture medium in increased amounts after rhIL-3 stimulation. On the basis of comigration with proteins in published 2-D gel electrophoresis databases and immunoblotting with a specific monoclonal antibody, we identified this protein as translationally controlled tumor protein (TCTP), also known as p23 or IgE-dependent histamine-releasing factor. The antibody was shown to be specific for TCTP/IgE-dependent histamine-releasing factor by blotting on 2-D gels of proteins from human lymphocytes and the human basophilic cell line KU812, followed by N-terminal amino-acid sequencing of the bound protein. Densitometric analysis of the gels showed that the synthesis of IgE-dependent histamine-releasing factor in human basophil leukocytes was dose dependent upon rhIL-3 stimulation with an optimum of 100 ng/ml. The level of the protein in the medium was also highest at an optimal rhIL-3 concentration of 100 ng/ml. Supernatants from cultured basophils were able to stimulate histamine release from other basophils. This histamine release was decreased by precipitation of TCTP/IgE-dependent histamine-releasing factor from these supernatants.
Nielsen
H V
HV
Department of Pediatrics, Aarhus University Hospital, Denmark.
Johnsen
A H
AH
Sanchez
J C
JC
Hochstrasser
D F
DF
Schiøtz
P O
PO
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001491:Basophils / Q000187:drug effects / Q000378:metabolism*; D014408:Biomarkers, Tumor; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D006636:Histamine Release / Q000502:physiology*; D006801:Humans; D015151:Immunoblotting; D007377:Interleukin-3 / Q000494:pharmacology*; D008222:Lymphokines / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000494:pharmacology; D000090324:Tumor Protein, Translationally-Controlled 1
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D007377:Interleukin-3; D008222:Lymphokines; D011994:Recombinant Proteins; C000715647:TPT1 protein, human; D000090324:Tumor Protein, Translationally-Controlled 1