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首页> 外文期刊>The Journal of Immunology: Official Journal of the American Association of Immunologists >Gamma-irradiation directly affects the formation of coding joints in SCID cell lines.
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Gamma-irradiation directly affects the formation of coding joints in SCID cell lines.

机译：伽马射线辐射直接影响SCID细胞系中编码接头的形成。

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SCID mice have a defect in the catalytic subunit of the DNA-dependent protein kinase, causing increased sensitivity to ionizing radiation in all tissues and severely limiting the development of B and T cell lineages. SCID T and B cell precursors are unable to undergo normal V(D)J recombination: coding joint and signal joint products are less frequently formed and often will exhibit abnormal structural features. Paradoxically, irradiation of newborn SCID mice effects a limited rescue of T cell development. It is not known whether irradiation has a direct impact on the process of V(D)J joining, or whether irradiation of the thymus allows the outgrowth of rare recombinants. To investigate this issue, we sought to demonstrate an irradiation effect ex vivo. Here we have been able to reproducibly detect low-frequency coding joint products with V(D)J recombination reporter plasmids introduced into SCID cell lines. Exposure of B and T lineage cells to 100 cGy of gamma irradiation made no significant difference with respect to the number of coding joint and signal joint recombination products. However, in the absence of irradiation, the coding joints produced in SCID cells had high levels of P nucleotide insertion. With irradiation, markedly fewer P insertions were seen. The effect on coding joint structure is evident in a transient assay, in cultured cells, establishing that irradiation has an immediate impact on the process of V(D)J recombination. A specific proposal for how the DNA-dependent protein kinase catalytic subunit influences the opening of hairpin DNA intermediates during coding joint formation in V(D)J recombination is presented.
机译：SCID小鼠的DNA依赖性蛋白激酶催化亚基有缺陷，导致所有组织对电离辐射的敏感性增加，并严重限制了B和T细胞谱系的发育。 SCID T和B细胞前体不能正常进行V（D）J重组：编码接头和信号接头产物的形成频率较低，并且经常表现出异常的结构特征。矛盾的是，新生SCID小鼠的辐射影响了T细胞发育的有限挽救。尚不清楚辐射是否直接影响V（D）J的结合过程，或者胸腺的辐射是否会导致稀有重组体的生长。为了研究这个问题，我们试图证明离体的辐射效果。在这里，我们已经能够用引入SCID细胞系的V（D）J重组报告质粒可再现地检测低频编码联合产物。 B和T谱系细胞暴露于100 cGy的γ射线照射下，编码接头和信号接头重组产物的数量没有显着差异。然而，在没有照射的情况下，SCID细胞中产生的编码接头具有高水平的P核苷酸插入。照射后，可见到明显更少的P插入。在瞬时分析中，在培养的细胞中，对编码关节结构的影响是显而易见的，从而确定辐照对V（D）J重组过程具有直接影响。提出了有关DNA依赖性蛋白激酶催化亚基如何在V（D）J重组的编码接头形成过程中影响发夹DNA中间体打开的具体建议。

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