TY - JOUR
T1 - Generation of human monoclonal antibodies by transformation of lymphoblastoid B cells with ras oncogene
AU - Shammah, Suzy
AU - Liliana Mantovani, T.
AU - Dalla-Favera, Riccardo
AU - Casali, Paolo
N1 - Funding Information:
Correspondence to: S. Shammah, Division of Oncology, Department of Pathology, College of Physicians and Surgeons, Columbia University, 630 W 168 Street, New York, NY 10032, USA. Tel.: (212) 305-6048; Fax: (212) 305-5498. * This work was supported by the U.S.P.H.S. (N.I.H.) grants CA-44029 (to R.D.F.), AR-40908 and CA-16087 (to P.C.) and by a grant from the BASF Bioresearch Corporation (to P.C.). S.S. was supported by a Fellowship from the Istituto Nazionale Tumori, Milan, Italy; T.L.M. was supported by Italfarmaco S.p.A., Milan, Italy; P.C. is a Kaplan Cancer Scholar. Abbreviations: hu-mAb, human monoclonal antibody; EBV, Epstein-Barr virus; LC, lymphoblastoid cell; neo, neomicine; hyb, hybridoma; ras-LC, ras-transformed LC; neo-LC, control-infected LC; hyb-LC, LC fused with a somatic cell hybridization partner.
PY - 1993
Y1 - 1993
N2 - 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.
AB - 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.
KW - Lymphoblastoid cell line
KW - Monoclonal antibody
KW - Ras oncogene
KW - Retroviral vector
KW - human
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U2 - 10.1016/0022-1759(93)90004-Q
DO - 10.1016/0022-1759(93)90004-Q
M3 - Article
C2 - 8450236
AN - SCOPUS:0027478974
VL - 160
SP - 19
EP - 25
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1
ER -