Distinctive High Expression of Antiretroviral APOBEC3 Protein in Mouse Germinal Center B Cells.
Distinctive High Expression of Antiretroviral APOBEC3 Protein in Mouse Germinal Center B Cells.
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DOI：
10.3390/v14040832
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发表时间：
2022-04-17
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影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
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限制病毒的APOBEC3在蛋白水平上的组织和亚细胞定位及其在细胞活化时的变化对于理解这种胞苷脱氨酶的生理功能很重要，但尚未在体内进行彻底的分析。为了精确跟踪APOBEC3蛋白在不同小鼠组织和细胞群中可能的激活诱导表达水平变化，我们利用基因组编辑技术建立了带框架内FLAG标签的表达APOBEC3蛋白的敲入小鼠。在免疫刺激前后分别进行流式细胞术和免疫组织化学分析。培养的B细胞表达APOBEC3蛋白水平高于T细胞。新制备的B细胞的所有分化和激活阶段都表达了显著水平的APOBEC3蛋白，但生发中心细胞在其细胞质中具有最高水平的APOBEC3蛋白。在体内用绵羊红细胞免疫刺激后，APOBEC3蛋白高表达的生发中心细胞数量增加，但每个细胞中flag特异性荧光强度没有变化。T细胞，即使是生发中心的T细胞，也没有表达显著水平的APOBEC3蛋白。因此，小鼠APOBEC3蛋白在生发中心B细胞中有明显的高水平表达。尽管生发中心细胞数量增加，但抗原刺激并未影响细胞APOBEC3蛋白的表达水平。
Tissue and subcellular localization and its changes upon cell activation of virus-restricting APOBEC3 at protein levels are important to understanding physiological functions of this cytidine deaminase, but have not been thoroughly analyzed in vivo. To precisely follow the possible activation-induced changes in expression levels of APOBEC3 protein in different mouse tissues and cell populations, genome editing was utilized to establish knock-in mice that express APOBEC3 protein with an in-frame FLAG tag. Flow cytometry and immunohistochemical analyses were performed prior to and after an immunological stimulation. Cultured B cells expressed higher levels of APOBEC3 protein than T cells. All differentiation and activation stages of freshly prepared B cells expressed significant levels of APOBEC3 protein, but germinal center cells possessed the highest levels of APOBEC3 protein localized in their cytoplasm. Upon immunological stimulation with sheep red blood cells in vivo, germinal center cells with high levels of APOBEC3 protein expression increased in their number, but FLAG-specific fluorescence intensity in each cell did not change. T cells, even those in germinal centers, did not express significant levels of APOBEC3 protein. Thus, mouse APOBEC3 protein is expressed at distinctively high levels in germinal center B cells. Antigenic stimulation did not affect expression levels of cellular APOBEC3 protein despite increased numbers of germinal center cells.
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影响因子：
6.7
作者：
Takamura S;Kajiwara E;Tsuji-Kawahara S;Masumoto T;Fujisawa M;Kato M;Chikaishi T;Kawasaki Y;Kinoshita S;Itoi M;Sakaguchi N;Miyazawa M
通讯作者：
Miyazawa M
影响因子：
5.6
作者：
Lackey, Lela;Demorest, Zachary L.;Land, Allison M.;Hultquist, Judd F.;Brown, William L.;Harris, Reuben S.
通讯作者：
Harris, Reuben S.
影响因子：
5.4
作者：
Koning, Fransje A.;Newman, Edmund N. C.;Malim, Michael H.
通讯作者：
Malim, Michael H.
影响因子：
14.9
作者：
Drozdetskiy A;Cole C;Procter J;Barton GJ
通讯作者：
Barton GJ
影响因子：
5.8
作者：
Granadillo Rodríguez M;Flath B;Chelico L
通讯作者：
Chelico L