Scharsack J P, Steinhagen D, Körting W, Leibold W, Schuberth H J
Fish Disease Research Unit, School of Veterinary Medicine, Hannover, Germany.
Dis Aquat Organ. 2000 Jul 14;41(3):203-10. doi: 10.3354/dao041203.
The activation of carp peripheral blood leukocytes (PBL) was analysed radiometrically and by means of flow cytometry (FCM) in order to compare the results obtained with both methods. The qualitative and quantitative FCM analyses of cellular morphology and viability resulted in a further characterisation of proliferative responses of carp PBL to Trypanoplasma borreli in vivo and in vitro. The lymphocyte population of PBL from T. borreli-infected carp exhibited a marked shift in forward scattered light (FSC; cell size). When PBL from healthy carp were stimulated with mitogens in vitro, a lymphoid population with increased FSC profiles was also observed. The number of these cells coincided to ratios of 3H-thymidine incorporation, recorded from corresponding cultures. Thus, it was concluded that the increase in size of stimulated lymphocytes could be due to blastogenic transformation. The advantage of the FCM procedure is that activation and proliferation of carp lymphocytes can be monitored without labelling the cells. Cocultures of mitogen-stimulated carp PBL and T. borreli revealed the ability of the parasite to suppress lymphocyte proliferation in vitro.
为比较两种方法的结果，采用放射测量法和流式细胞术（FCM）对鲤鱼外周血白细胞（PBL）的激活情况进行了分析。对细胞形态和活力进行的FCM定性和定量分析，进一步明确了鲤鱼PBL在体内和体外对博氏锥虫的增殖反应特征。来自感染博氏锥虫鲤鱼的PBL淋巴细胞群体在前向散射光（FSC；细胞大小）方面表现出明显变化。当体外用有丝分裂原刺激健康鲤鱼的PBL时，也观察到FSC图谱增加的淋巴细胞群体。这些细胞的数量与相应培养物中记录的3H - 胸腺嘧啶核苷掺入率相符。因此，得出结论，受刺激淋巴细胞大小的增加可能是由于母细胞转化。FCM方法的优点是无需标记细胞就能监测鲤鱼淋巴细胞的激活和增殖。有丝分裂原刺激的鲤鱼PBL与博氏锥虫的共培养揭示了该寄生虫在体外抑制淋巴细胞增殖的能力。
