Human monoclonal thyroglobulin autoantibodies of high affinity. I. Production, characterisation and interaction with murine monoclonal thyroglobulin antibodies.
Four hybridomas secreting human thyroglobulin (Tg) autoantibodies of different IgG subclasses and light chain types (IgG1 lambda, IgG1 kappa, IgG2 lambda and IgG2 kappa) were obtained by direct fusion of Hashimoto thyroid lymphocytes with the mouse myeloma X63-Ag.653. The autoantibodies were specific for human Tg and the functional affinities were high (only 2.6-3.9 log10 pM Tg required to give 50% inhibition of binding in ELISA). Using thyroid lymphocytes, 4 lines secreting Tg autoantibodies were obtained from 11 fusions compared with 1 line from 32 fusions of Epstein Barr virus infected blood lymphocytes, which emphasises the importance of using lymphocytes derived from a tissue known to be enriched in thyroid autoantibody secreting precursor B cells. These 4 human Tg autoantibodies, as well as an IgG2 lambda Tg antibody previously derived from Hashimoto blood B cells and an IgG4 kappa monoclonal Tg antibody present in a Hashimoto serum, were used in attempts to probe the interaction between human Tg autoantibodies and the Tg molecule (2 polypeptides of 330 KD). The binding to 125-I Tg by 3/7 murine monoclonal antibodies was inhibited (36-78%) by an IgG2 lambda and an IgG4 kappa human monoclonal Tg autoantibody, indicating an overlap between the epitopes recognised by these 3 murine monoclonal Tg antibodies and 2 monoclonal human Tg autoantibodies. None of the human Tg autoantibodies (or the murine monoclonal Tg antibodies) bound to Tg denatured by reduction and alkylation. Although the number of observations is limited, our study demonstrates that high affinity human monoclonal Tg autoantibodies, like polyclonal serum Tg autoantibodies, recognise non-linear B cell epitopes on conformationally intact human Tg.
Fukuma
N
N
Department of Medicine, University of Wales College of Medicine, Heath Park, Cardiff, U.K.
Petersen
V B
VB
McLachlan
S M
SM
Pegg
C A
CA
Rees Smith
B
B
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000915:Antibody Affinity; D001323:Autoantibodies / Q000737:chemistry*; D000939:Epitopes / Q000737:chemistry*; D006111:Graves Disease / Q000097:blood / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D007453:Iodide Peroxidase / Q000276:immunology; D051379:Mice; D013954:Thyroglobulin / Q000276:immunology*; D013961:Thyroid Gland / Q000276:immunology*; D013967:Thyroiditis, Autoimmune / Q000097:blood / Q000276:immunology
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D000939:Epitopes; D013954:Thyroglobulin; D007453:Iodide Peroxidase