Human platelet-derived growth factor: radioimmunoassay and discovery of a specific plasma-binding protein.
The platelet-derived growth factor (PDGF) is the principal mitogen in serum for cultured cells of mesenchymal origin. PDGF also is a potent chemotactic protein for inflammatory cells and for cells required for wound repair. Because activity levels of PDGF in biological fluids are difficult to measure, we attempted to develop a radioimmunoassay for PDGF. Rabbits were immunized with purified PDGF; the antiserum obtained was monospecific for PDGF in immunodiffusion analysis against concentrated platelet lysates, serum, and plasma. A radioimmunoassay for PDGF was developed with a sensitivity of congruent to 0.2 ng/ml. Levels of PDGF in plasma/serum were measured and compared with PDGF levels determined by a receptor-competition assay and by a standard biological assay measuring incorporation of [3H]thymidine into 3T3 cells. Radioimmunoassay showed apparent PDGF levels of 50 ng/ml in human plasma and 103 ng/ml in serum. The 50 ng/ml PDGF in plasma was unexpected because the plasma samples contained little or no platelet release products as determined by very low levels of platelet factor 4. We therefore sought an immunologically reactive PDGF molecule in human plasma. No immunologically reactive protein was detected by immunodiffusion analysis or when plasma was treated with an immunoaffinity gel. Subsequently, a 125I-PDGF-binding protein was identified; the 125I-PDGF-plasma-binding protein complex was not reactive with anti-PDGF immunoglobulin. Correction for 125I-PDGF bound by the plasma-binding protein established serum levels of PDGF of congruent to 50 ng/ml; congruent to 50 ng/ml PDGF was found in serum by radioreceptor-competition assays and by mitogenic assays as well. The plasma-binding protein may serve to clear PDGF released in the circulation, thereby limiting PDGF activity to its local interactions at the site of blood-vessel injury.
D000818:Animals; D000918:Antibody Specificity; D001798:Blood Proteins / Q000302:isolation & purification; D006133:Growth Substances / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D051379:Mice; D010455:Peptides / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification*; D010982:Platelet-Derived Growth Factor; D011485:Protein Binding; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods*
D001798:Blood Proteins; D006133:Growth Substances; D010455:Peptides; D010982:Platelet-Derived Growth Factor