Immunochemical studies of antigenic lymphoma-associated proteins.
We have previously reported that plasma from patients with anaplastic lymphoma kinase (ALK)-positive lymphoma contains antibodies against the oncogenic kinase NPM-ALK protein characteristic of this disease. We investigated whether this reactivity represents a phenomenon unique to ALK-positive lymphoma by screening plasma from patients with follicular lymphoma for antibodies to BCL-2 protein. Eight out of 10 samples showed such reactivity (and in six cases gave specific staining of BCL-2-transfected cells). As these findings suggest a new biochemical approach to the identification of oncogenic proteins in lymphoma, we investigated whether antibodies present in patients with ALK-positive lymphoma can precipitate NPM-ALK in quantities which should be sufficient for further analysis. We found that plasma samples from all10 patients studied immunoprecipitated NPM-ALK asaprotein visible in silver-stained sodium dodecyl sulphatepolyacrylamide gels. Finally we demonstrated that NPM-ALK could be visualized more clearly if it were immunoprecipitated from extracts of cells in which newly synthesized proteins had been labelled with 35S and then identified by autoradiography. These results suggest a strategy for using patients' autoantibodies to screen for antibodies to other tumour-associated proteins.
Pulford
Karen
K
Nuffield Department of Clinical Laboratory Sciences, Leukaemia Research Fund Immunodiagnostics Unit, John Radcliffe Hospital, Oxford, UK. karen.pulford@ndcls.ox.ac.uk
Roberton
Helen
H
Banham
Alison H
AH
Hatton
Christian S R
CS
Mason
David Y
DY
D015295:Antigens, Tumor-Associated, Carbohydrate / Q000276:immunology*; D001323:Autoantibodies / Q000097:blood*; D001345:Autoradiography; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D019169:Jurkat Cells; D008224:Lymphoma, Follicular / Q000276:immunology*; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000032:analysis / Q000276:immunology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000276:immunology*; D013462:Sulfur Radioisotopes
D015295:Antigens, Tumor-Associated, Carbohydrate; D001323:Autoantibodies; D019253:Proto-Oncogene Proteins c-bcl-2; D013462:Sulfur Radioisotopes; C107023:p80(NPM-ALK) protein; D011505:Protein-Tyrosine Kinases