Dighiero G, Lymberi P, Holmberg D, Lundquist I, Coutinho A, Avrameas S
J Immunol. 1985 Feb;134(2):765-71.
Spleen cells from 6-day-old nonimmunized BALB/c and BALB.B10 mice were fused with the nonsecreting hybridoma cell line Sp2/0. Three hundred and eighty-four immunoglobulin-secreting hybrids were screened for antibody activity against mouse actin, tubulin, and myosin, and against TNP, peroxidase, renin, DNA, and neurofilaments. At least 24 hybridomas in the collection (6.25%) exhibited antibody activity against this panel of antigens. Ten of these hybrids were cloned, were propagated, and the corresponding monoclonal IgM protein was isolated from ascitic fluids and was further characterized. At least four groups of antibody specificities were identified: 1) one clone reacting with TNP only; 2) one clone reacting with both actin and tubulin; 3) two clones which bound to both TNP and actin; and 4) a fourth group, comprising the six other clones, which all exhibited widespread reactivity and bound to actin, tubulin, myosin, and TNP. These results indicate: 1) B cell clones directed against self antigens are activated in the internal environment and are recovered consequently by somatic cell hybridization; 2) the widespread antibody specificities found for these newborn mouse antibodies are very similar to those previously characterized with human natural antibodies and human monoclonal Ig; and 3) the frequency of B cells binding to cytoskeletal proteins and TNP is very high (at least 6.25%).
将6日龄未免疫的BALB/c和BALB.B10小鼠的脾细胞与非分泌性杂交瘤细胞系Sp2/0进行融合。对384个分泌免疫球蛋白的杂交瘤进行筛选，检测其针对小鼠肌动蛋白、微管蛋白和肌球蛋白，以及针对三硝基苯（TNP）、过氧化物酶、肾素、DNA和神经丝的抗体活性。该集合中的至少24个杂交瘤（6.25%）表现出针对这组抗原的抗体活性。其中10个杂交瘤被克隆、传代培养，从腹水液中分离出相应的单克隆IgM蛋白并进一步鉴定。鉴定出至少四组抗体特异性：1）一个克隆仅与TNP反应；2）一个克隆与肌动蛋白和微管蛋白都反应；3）两个克隆与TNP和肌动蛋白都结合；4）第四组包括其他六个克隆，它们都表现出广泛的反应性，与肌动蛋白、微管蛋白、肌球蛋白和TNP结合。这些结果表明：1）针对自身抗原的B细胞克隆在体内环境中被激活，并因此通过体细胞杂交被回收；2）这些新生小鼠抗体中发现的广泛抗体特异性与先前用人天然抗体和人单克隆Ig所鉴定的特异性非常相似；3）与细胞骨架蛋白和TNP结合的B细胞频率非常高（至少6.25%）。
