Diagnosis of t(2;5)(p23;q35)-associated Ki-1 lymphoma with immunohistochemistry.
Some Ki-1 lymphomas carry a specific chromosomal translocation, t(2;5)(p23;q35). We have recently found a novel hyperphosphorylated 80-kD protein tyrosine kinase, p80, in a human Ki-1 lymphoma with this translocation. Subsequent cDNA cloning showed that p80 is a fusion protein of two different genes on chromosome 2p23 and 5q35, the novel tyrosine kinase gene and nucleophosmin gene, respectively. In this study, we intended to detect p80 on lymphoma tissues with immunologic methods. Thus, we developed rabbit polyclonal antibody using a synthetic peptide corresponding to a part of its kinase domain. The antibody (anti-p80) immunoprecipitated and immunoblotted p80 specifically from AMS3. Then, to examine whether t(2;5)(p23;q35) was present on biopsied lymphomas, reverse transcriptase-polymerase chain reaction (RT-PCR) covering the fusion junction of p80 mRNA was performed. Among 10 Ki-1 lymphomas and 10 additional lymphomas other than the Ki-1 lymphomas, expression of p80 mRNA was detected in three cases exclusively. When these 20 cases and additional 30 lymphomas were immunostained with anti-p80, positive staining was noted exclusively in the three cases found by PCR to have harbored the p80 mRNA. Thus, the present immunostaining, as well as PCR, was shown to be efficient for detecting lymphomas producing this chimeric protein/mRNA.
Shiota
M
M
Department of Pathology and Oncology, Institute of Medical Science, University of Tokyo, Japan.
Fujimoto
J
J
Takenaga
M
M
Satoh
H
H
Ichinohasama
R
R
Abe
M
M
Nakano
M
M
Yamamoto
T
T
Mori
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002889:Chromosomes, Human, Pair 2 / Q000648:ultrastructure*; D002895:Chromosomes, Human, Pair 5 / Q000648:ultrastructure*; D003429:Cross Reactions; D006801:Humans; D007150:Immunohistochemistry; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology; D017728:Lymphoma, Large-Cell, Anaplastic / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics* / Q000276:immunology; D009687:Nuclear Proteins / Q000235:genetics*; D000090243:Nucleophosmin; D011505:Protein-Tyrosine Kinases / Q000235:genetics* / Q000276:immunology; D020794:Receptor Protein-Tyrosine Kinases / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology; D014178:Translocation, Genetic; D014407:Tumor Cells, Cultured
C000716967:NPM1 protein, human; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D000090243:Nucleophosmin; C489300:LTK protein, human; C064626:Ltk protein, mouse; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases