Generation of human monoclonal antibodies against HIV-1 proteins; electrofusion and Epstein-Barr virus transformation for peripheral blood lymphocyte immortalization.
Electrofusion and EBV transformation were studied by immortalizing human PBLs from blood of HIV-1-positive volunteers. A panel of 33 cell lines producing human monoclonal antibodies (Hu-MAbs) against HIV-1 was established by cell fusion or EBV transformation. For the first fusion experiments the source of B lymphocytes was peripheral blood of HIV-1-infected donors in CDC stages II or III with CD4 cell counts higher than 500/mm3. Later on, from these patients only, those with high anti-HIV titers were chosen as blood donors. By that means the yield of stable specific hybridomas was increased twofold. In our experiments electrofusion turned out to be a more efficient immortalization method than EBV transformation, due to a high and constant immortalization rate. The hybridomas were stable after intensive subcloning and could be cultivated over a period of 8 months without loss in monoclonal antibody production. Immunoglobulin class, subtype, reactivity against HIV-1 proteins, Western blot patterns, immunofluorescence, and epitopes were characterized. The subtype of all antibodies was IgG1 or IgG3. The light chain was predominantly kappa. All antibodies showed reactivity against HIV-1 envelope or core protein. All hybridomas were stable and suited for mass production. Several Hu-MAbs are becoming an important tool in the field of diagnosis, research, and immunotherapy.
Buchacher
A
A
Institute of Applied Microbiology, University of Foresty and Agriculture, Vienna, Austria.
Predl
R
R
Strutzenberger
K
K
Steinfellner
W
W
Trkola
A
A
Purtscher
M
M
Gruber
G
G
Tauer
C
C
Steindl
F
F
Jungbauer
A
A
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry; D001402:B-Lymphocytes / Q000276:immunology; D002459:Cell Fusion; D002460:Cell Line; D002472:Cell Transformation, Viral; D000939:Epitopes; D015483:HIV Antibodies / Q000096:biosynthesis* / Q000737:chemistry; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D015658:HIV Infections / Q000276:immunology; D015497:HIV-1 / Q000276:immunology*; D004854:Herpesvirus 4, Human; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007120:Immunochemistry; D066298:In Vitro Techniques; D009500:Neutralization Tests
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120