EFFICIENCY OF TRANSFORMATION OF LYMPHOCYTES BY EPSTEIN-BARR VIRUS

EFFICIENCY OF TRANSFORMATION OF LYMPHOCYTES BY EPSTEIN-BARR VIRUS
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DOI:
10.1016/0042-6822(77)90292-6
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发表时间:
1977-01-01
期刊:
影响因子:
3.7
通讯作者:
HESTON, L
HESTON, L
中科院分区:
医学3区
文献类型:
--
作者:
HENDERSON, E;MILLER, G;HESTON, L

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研究了宿主细胞和病毒因素对淋巴细胞对EB病毒永生化易感性的影响。EB病毒B 95 -8株的永生化遵循一击动力学。采用有限稀释技术,当在人脐带白细胞(HUCL)上测定时,约有1-20个病毒颗粒发生转化。来自成年人和绒猴血液的混合单核白细胞对永生化的敏感性分别比HUCL低100倍和1000倍。用植物血凝素预处理脐带血白细胞可使其转化性增加50%。来自大肠杆菌的脂多糖(LPS)使观察到的转化事件的数量增加300- 500%。LPS似乎通过刺激细胞DNA合成以外的机制增强转化。采用自体淋巴细胞或人胎盘细胞作为饲养细胞的转化中心试验用于确定永生化脐带细胞的数量。在30-100个颗粒/细胞的感染多重性下,几乎所有易感细胞都饱和的多重性,约1/200至1/500的混合单核细胞转化。当混合细胞群中T [胸腺衍生]淋巴细胞耗尽时,转化细胞的数量增加了4倍。当对转化的脐带血白细胞的平板接种效率进行校正时,计算出至少10%的病毒暴露细胞建立了永久细胞系。这个数字可能仍然是一个低估,因为一个人口组成的全部易感细胞尚未使用。
Host cell and viral factors which influence the susceptibility of lymphocytes to immortalization by the Epstein-Barr virus (EBV) were studied. Immortalization by the B95-8 strain of EBV follows one-hit kinetics. With a limiting dilution technique, approximately 1-20 virus particles is transforming when assayed on human umbilical cord leukocytes (HUCL). Mixed mononuclear leukocytes from adult human and marmoset blood are 100- and 1000-fold, respectively, less sensitive to immortalization than HUCL. Pretreatment of cord blood leukocytes with phytohemagglutinin results in a 50% increase in their transformability. Lipopolysaccharide (LPS) from Escherichia coli increases the number of transformation events observed by 300-500%. LPS appears to enhance transformation by mechanisms other than stimulation of cellular DNA synthesis. A transformed center assay employing either autochthonous lymphocytes or human placental cells as feeders was used to determine the number of umbilical cord cells which were immortalized. At multiplicities of infection of 30-100 particles/cell, multiplicities which saturate nearly all the susceptible cells, approximately 1 in 200 to 1 in 500 mixed mononuclear cells transformed. When the mixed population of cells was depleted of T [thymus-derived] lymphocytes, there was a 4-fold increase in the number of transformed cells. When correction is made for the plating efficiency of transformed cord blood leukocytes, it is calculated that at least 10% of virus-exposed cells establish permanent lines. This number is probably still an underestimate since a population which is composed entirely of susceptible cells was not yet used.