Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 19-25 |
| Number of pages | 7 |
| Journal | Journal of Immunological Methods |
| Volume | 160 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 1 1993 |
| Externally published | Yes |
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Keywords
- human
- Lymphoblastoid cell line
- Monoclonal antibody
- Ras oncogene
- Retroviral vector
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology
Cite this
Generation of human monoclonal antibodies by transformation of lymphoblastoid B cells with ras oncogene. / Shammah, Suzy; Liliana Mantovani, T.; Dalla-Favera, Riccardo; Casali, Paolo.
In: Journal of Immunological Methods, Vol. 160, No. 1, 01.01.1993, p. 19-25.Research output: Contribution to journal › Article
}
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
PY - 1993/1/1
Y1 - 1993/1/1
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 - human
KW - Lymphoblastoid cell line
KW - Monoclonal antibody
KW - Ras oncogene
KW - Retroviral vector
UR - http://www.scopus.com/inward/record.url?scp=0027478974&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027478974&partnerID=8YFLogxK
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 -