Generation of human monoclonal antibodies by transformation of lymphoblastoid B cells with ras oncogene.
Human monoclonal antibodies (hu-mAbs) of predetermined specificity and isotype are potentially important for a variety of applications, including therapy and diagnosis. Their efficient generation, however, is still hampered by technical difficulties. Even the most established approaches to the generation of hu-mAbs, i.e., B cell immortalization by Epstein-Barr virus (EBV) and/or fusion with appropriate myeloma cell lines, are characterized by a relatively low efficiency. It has been shown that expression of activated Ha- or N-ras oncogenes causes the malignant transformation and plasmacytoid differentiation of EBV-immortalized lymphoblastoid cell (LC) lines, suggesting that activated ras oncogenes can convert LC lines into effective hu-mAb producers. We have used retroviral vector-mediated gene transfer to introduce an activated Ha-ras (v-ras) oncogene into four distinct LC lines producing hu-mAbs of different classes (IgM and IgG) and specificities (to human insulin, human thyroglobulin and rabies virus glycoprotein). The cloning efficiency and antibody secretion of these ras-transformed LC (ras-LC) lines were compared with those of the hybrid LC (hyb-LC) lines generated by fusing the same parental LC lines with the Ig non-secretor F3B6 human-mouse hybrid cells. ras-LC lines were comparable to their hybrid counterparts in either parameter tested. This, together with the relatively higher efficiency of the method, suggests that ras transformation may constitute a valid alternative to the currently available technologies for hu-mAbs production from LC lines.
Shammah
S
S
Department of Pathology, College of Physicians and Surgeons, Columbia University, New York, NY 10032.
Mantovani
T L
TL
Dalla-Favera
R
R
Casali
P
P
D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000956:Antigens, Viral; D001402:B-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D002472:Cell Transformation, Viral / Q000276:immunology; D015870:Gene Expression; D011905:Genes, ras / Q000235:genetics / Q000276:immunology; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D007330:Insulin Antibodies / Q000276:immunology*; D015689:Oncogene Protein p21(ras) / Q000235:genetics / Q000276:immunology*; D012333:RNA, Messenger / Q000235:genetics; D012190:Retroviridae; D013954:Thyroglobulin / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D006023:Glycoproteins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007330:Insulin Antibodies; D012333:RNA, Messenger; D014759:Viral Envelope Proteins; C064951:glycoprotein G, Rabies virus; D013954:Thyroglobulin; D015689:Oncogene Protein p21(ras)