Kojima K, Matsuyama T, Fukumori J, Matsuzaki I, Tanaka H
Nihon Naibunpi Gakkai Zasshi. 1985 Mar 20;61(3):162-7. doi: 10.1507/endocrine1927.61.3_162.
We established an anti-thyroglobulin antibody-producing cell line using the peripheral lymphocytes from a patient with chronic thyroiditis. The method was based on preselection by "panning", transformation by EB virus and twice clonings by "limiting dilution". The cells of the cloned cell line (Yo3CTX10) had neither E-receptor nor IgG X Fc-receptor but had C3-receptor as shown by rosetting. We could not detect surface immunoglobulin, but we could detect cytoplasmic immunoglobulin (IgG lambda) with FITC-stainings. This cell line has continuously secreted anti-thyroglobulin antibody for 8 months, which was IgG lambda as shown by immunoelectrophoresis and solid phase radioimmunoassay. The pattern obtained using the purified antibody showed two sharp bands (H, L-chains) in SDS polyacrylamide gel electrophoresis, but it showed six sharp bands in isoelectric focusing electrophoresis. Thus we could establish an oligoclonal cell line producing IgG lambda anti-thyroglobulin antibody.
我们利用一名慢性甲状腺炎患者的外周淋巴细胞建立了一个产生抗甲状腺球蛋白抗体的细胞系。该方法基于“淘选”进行预选、通过EB病毒转化以及通过“有限稀释”进行两次克隆。如玫瑰花结试验所示,克隆细胞系(Yo3CTX10)的细胞既没有E受体也没有IgG X Fc受体,但有C3受体。我们检测不到表面免疫球蛋白,但通过异硫氰酸荧光素染色能够检测到细胞质免疫球蛋白(IgGλ)。该细胞系连续8个月分泌抗甲状腺球蛋白抗体,免疫电泳和固相放射免疫测定显示其为IgGλ。使用纯化抗体得到的图谱在SDS聚丙烯酰胺凝胶电泳中显示出两条清晰的条带(重链、轻链),但在等电聚焦电泳中显示出六条清晰的条带。因此,我们能够建立一个产生IgGλ抗甲状腺球蛋白抗体的寡克隆细胞系。