Oates K K, Naylor P H, Goldstein A L
George Mason University, Fairfax, Virginia 20030.
Hybridoma. 1987 Feb;6(1):47-59. doi: 10.1089/hyb.1987.6.47.
The thymus is an endocrine organ which modulates T-cell immunity through the production of protein like peptides such as the thymosins. Thymosin alpha 1 was the first biologically active peptide isolated and sequenced from the partially purified thymic preparation, thymosin fraction 5, and has been extensively studied. Using synthetic Thymosin alpha 1, a heterologous rabbit antiserum has been raised and a radioimmunoassay has been developed. Although thymosin alpha 1 antibodies have been used in several histological studies, their use is limited by potential nonspecific cross-reactivities, unpredictable heterogenicity, variable affinities, and a limited unstandardized supply. In the studies, reported here, eight anti-thymosin alpha 1 monoclonal antibodies (MAbs) were produced by somatic cell fusion between spleen cells from immunized BALB/c mice and P3x64 Ag8.653 myeloma cells. The MAbs were screened for anti-thymosin alpha 1 specificity in a solid phase ELISA and a liquid phase RIA. Only those clones which secreted specific antibody as detected by both procedures were characterized for their heavy chain class and epitope specificity. The anti-thymosin alpha 1 monoclonal antibodies were then used for indirect immune fluorescence studies of perfused rat thymus. Thymosin alpha 1 containing cells were found primarily in the thymic medulla, confirming previous studies using the heterologous antisera. These studies demonstrated the specificity of the anti-thymosin alpha 1 monoclonal antibodies for immunochemical studies of intra- and extra-thymic localization of thymosin alpha 1. They also provide an important reagent for biological studies of the role of thymosin alpha 1, in vitro and in vivo.
胸腺是一种内分泌器官，它通过产生诸如胸腺素等蛋白质样肽来调节T细胞免疫。胸腺素α1是从部分纯化的胸腺制剂（胸腺素组分5）中分离并测序的首个具有生物活性的肽，并且已得到广泛研究。利用合成的胸腺素α1，制备了一种异源兔抗血清，并开发了一种放射免疫测定法。尽管胸腺素α1抗体已用于多项组织学研究，但其应用受到潜在的非特异性交叉反应、不可预测的异质性、可变亲和力以及供应未标准化且有限等因素的限制。在本文报道的研究中，通过免疫的BALB/c小鼠的脾细胞与P3x64 Ag8.653骨髓瘤细胞进行体细胞融合，产生了8种抗胸腺素α1单克隆抗体（MAb）。通过固相ELISA和液相RIA筛选这些MAb的抗胸腺素α1特异性。只有那些在两种检测方法中均检测到分泌特异性抗体的克隆，才对其重链类别和表位特异性进行表征。然后将抗胸腺素α1单克隆抗体用于灌注大鼠胸腺的间接免疫荧光研究。发现含胸腺素α1的细胞主要位于胸腺髓质，这证实了先前使用异源抗血清的研究。这些研究证明了抗胸腺素α1单克隆抗体在胸腺素α1胸腺内和胸腺外定位的免疫化学研究中的特异性。它们还为体外和体内研究胸腺素α1的作用提供了一种重要试剂。
