<原著>抗ヒトサイログロブリン単クローン抗体の基礎的研究およびその臨床応用(2) : 単クローン抗体を用いた各種甲状腺疾患におけるサイログロブリンエピトープの研究

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Nine murine monoclonal antibodies against human thyroglobulin (Tg) were prepared by the hybridoma technique. Polyclonal antibody to human Tg was produced in mice by ordinary immunization with human Tg. By using these anti-Tg antibodies, the reactivity with human Tgs, in 29 preparations obtained from thyroid cancer, thyroid adenoma, Graves' thyroid and healthy thyroid tissue, was investigated to elucidate the possible difference of epitope structures in Tg molecule. In enzyme-linked immunosorbent assay (ELISA), polyclonal anti-Tg antibody showed homogeneous reactivity to all of the Tg preparations, while the 9 monoclonal antibodies, named 1E3,2A1,2B3,2F4,3C6,4B9,4F1,7B6 and F8A4,were heterogeneous in the reactivity to Tgs. The monoclonal antibody, 2B3,bound well with normal Tgs but poorly with Tgs obtained from Graves thyroid, thyroid adenoma and thyroid cancer with the lowest affinity to cancer or adenoma-Tgs. For the other 8 monoclonal antibodies, the binding of each antibody with Tg did not show definitive differences in the source of Tg. In immuno-peroxidase staining of thyroid cancer using polyclonal and monoclonal antibodies, the colloid was stained stronger than the tumor cells when the polyclonal antibody and the 7 monoclonal antibodies other than 2B3 and 2F4 were employed ; the colloid and tumor cells were stained equally by 2F4 ; as for 2B3,the colloid was scarcely stained, although the tumor cells were stained markedly. This antibody, 2B3,was unique in that it did not stain the colloid in thyroid adenoma as well as in the cancer but stained the colloid of normal thyroid tissue. After the treatment with neuraminidase of the paraffin embedded section of thyroid adenoma, the tissue cells turned out to be positive for staining with 2B3. This suggests that some epitopes of Tg may exist as cryptic antigens covered with sialic acid in thyroid adenoma. Autoantibody to human Tg (HATg) was purified from the serum of patients with Hashimoto's thyroiditis. Two (3C6 and 7B6) out of the 9 monoclonal antibodies were shown to possess cross-reactivity with HATg in the study of antibody competition to Tg. Also, in similar competition tests using horse-radish peroxidase (HRPO)- labelled monoclonal antibodies and unlabelled monoclonal antibodies, the 9 monoclonal antibodies were classified into 7 clusters. Therefore, at least 7 different epitopes in Tg molecule were recognized by the 9 monoclonal antibodies obtained in the present study.
近畿大学の論文
1987-03-25

<原著>抗ヒトサイログロブリン単クローン抗体の基礎的研究およびその臨床応用(2) : 単クローン抗体を用いた各種甲状腺疾患におけるサイログロブリンエピトープの研究

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