Kligerman A D, Erexson G L, Wilmer J L
Basic Life Sci. 1984;29 Pt B:569-84. doi: 10.1007/978-1-4684-4892-4_6.
Peripheral blood lymphocytes (PBLs) offer many advantages for in vivo cytogenetic studies. They can be removed nonlethally from the animal allowing a subject to serve as its own control, permitting the analysis of cytogenetic damage over time. Furthermore, mature PBLs normally do not divide, and some populations are long-lived. Thus, they have the potential to accumulate DNA lesions during chronic exposures to genotoxicants. We have developed standard methodologies for the whole blood culture of rat and mouse PBLs to serve as models for determining the sensitivity of PBLs to cytogenetic damage. The cultures obtained with these protocols give reproducible results with high mitotic indices, stable baseline sister chromatid exchange (SCE) frequencies, and ample numbers of first-and second-division methaphases for scoring both chromosomal aberrations and SCEs. The methodologies have been especially useful for examining cytogenetic damage after inhalation exposures to toxicants such as ethylene oxide, formaldehyde, benzene, and nitrobenzene. Of these compounds, only benzene and ethylene oxide were found to induce significant dose-dependent increases in SCEs in PBLs. Also, dose-response curves have been obtained for several carcinogens administered by ip injection. These studies show that PBLs are sensitive indicators of the genotoxic effects of the carcinogens benzo (a)pyrene, 2-acetylaminofluorene, cyclophosphamide (CP), N-nitrosomorpholine, and ethylmethanesulfonate (EMS). In addition, because subpopulations of lymphocytes can be stimulated to divide using different mitogens, it has been possible to compare the sensitivity of murine B and T lymphocytes following in vitro and in vivo cyclophosphamide exposure. Once the sensitivity and selectivity of rodent lymphocyte cultures are determined, these assays should be valuable not only as a means for predicting which environmental agents could lead to increases in human cytogenetic damage, but also as a way to corroborate human cytogenetic studies.
外周血淋巴细胞（PBLs）为体内细胞遗传学研究提供了许多优势。可以在不致死的情况下从动物体内采集，使实验对象能够作为自身对照，从而可以分析随时间推移的细胞遗传学损伤。此外，成熟的PBLs通常不分裂，且某些群体寿命较长。因此，它们在长期暴露于遗传毒性物质期间有积累DNA损伤的潜力。我们已开发出大鼠和小鼠PBLs全血培养的标准方法，作为确定PBLs对细胞遗传学损伤敏感性的模型。使用这些方案获得的培养物可产生可重复的结果，具有高有丝分裂指数、稳定的基线姐妹染色单体交换（SCE）频率，以及足够数量的第一次和第二次分裂中期相，可用于对染色体畸变和SCE进行评分。这些方法对于检查吸入环氧乙烷、甲醛、苯和硝基苯等毒物后的细胞遗传学损伤特别有用。在这些化合物中，仅发现苯和环氧乙烷能诱导PBLs中SCEs显著的剂量依赖性增加。此外，还获得了几种经腹腔注射给药的致癌物的剂量反应曲线。这些研究表明，PBLs是致癌物苯并（a）芘、2-乙酰氨基芴、环磷酰胺（CP）、N-亚硝基吗啉和甲磺酸乙酯（EMS）遗传毒性作用的敏感指标。此外，由于可以使用不同的有丝分裂原刺激淋巴细胞亚群进行分裂，因此有可能比较体外和体内环磷酰胺暴露后小鼠B淋巴细胞和T淋巴细胞的敏感性。一旦确定了啮齿动物淋巴细胞培养的敏感性和选择性，这些检测不仅作为预测哪些环境因素可能导致人类细胞遗传学损伤增加的手段有价值，而且作为证实人类细胞遗传学研究的一种方法也有价值。
