|Title||Human monoclonals from antigen-specific selection of B lymphocytes and transformation by EBV.|
|Publication Type||Journal Article|
|Year of Publication||1986|
|Authors||Casali P, Inghirami G, Nakamura M, Davies TF, Notkins AL|
|Date Published||1986 Oct 24|
|Keywords||Antibodies, Monoclonal, B-Lymphocytes, Cell Separation, Cell Transformation, Viral, Diabetes Mellitus, Type 1, Flow Cytometry, Herpesvirus 4, Human, Humans, Immunoglobulin M, Receptors, Antigen, B-Cell, Tetanus Toxoid, Thyroglobulin, Thyroiditis, Autoimmune|
Hybridoma technology has made it possible to prepare monoclonal antibodies with the use of murine lymphocytes. Attempts to extend this technology to the human level, however, have met with difficulties. A method has been developed for making human monoclonal antibodies of predetermined specificity. Biotinylated antigens (human thyroglobulin or tetanus toxoid) were incubated with human B lymphocytes from peripheral blood. The lymphocytes to which the antigens bound were selected by fluorescence-activated cell sorting. Positively selected (high fluorescence) and negatively selected (low fluorescence) cells were then transformed with Epstein-Barr virus (EBV) and grown in microculture wells. All wells from the positively selected fraction produced antigen-specific antibody (95 to 1800 nanogram-equivalents per milliliter), whereas fewer than 6% of the wells from negatively selected fraction made any detectable antibody (less than 10 nanogram-equivalents per milliliter). When the positively selected EBV-transformed cells were cultured in limiting dilution, clones were obtained that made antigen-specific monoclonal antibodies. By this method, monoclonal antibodies to both foreign antigens and autoantigens can be prepared from the normal human B-cell repertoire.
Related Faculty:Giorgio Inghirami, M.D.