Estimation of bovine leukemia virus (BLV) proviral load harbored by lymphocyte subpopulations in BLV-infected cattle at the subclinical stage of enzootic bovine leucosis using BLV-CoCoMo-qPCR.
Estimation of bovine leukemia virus (BLV) proviral load harbored by lymphocyte subpopulations in BLV-infected cattle at the subclinical stage of enzootic bovine leucosis using BLV-CoCoMo-qPCR.
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DOI：
10.1186/1746-6148-9-95
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发表时间：
2013-05-04
影响因子：
2.6
通讯作者：
Aida Y
中科院分区：
文献类型：
--
作者：
Panei CJ;Takeshima SN;Omori T;Nunoya T;Davis WC;Ishizaki H;Matoba K;Aida Y
牛白血病病毒（BLV）与牛地方性白血病（EBL）有关，EBL是牛最常见的肿瘤性疾病。BLV感染可能在无白细胞（AL）阶段保持临床沉默，引起持续性淋巴细胞增多（PL），或更罕见的B细胞淋巴瘤。已在未患肿瘤的感染牛的B细胞、CD 2 + T细胞、CD 3 + T细胞、CD 4 + T细胞、CD 8 + T细胞、γ/δ T细胞、单核细胞和粒细胞中鉴定出BLV，尽管最持续感染的细胞是CD 5 + B细胞。BLV引起不受控制的CD 5 + B细胞增殖的机制尚不清楚。最近，我们开发了一种新的定量实时聚合酶链反应（PCR）方法，BLV-CoCoMo-qPCR，这使我们能够证明，前病毒载量不仅与BLV感染，评估合胞体形成，但也与BLV疾病进展。本研究报告了通过细胞分选和BLV-CoCoMo-qPCR检测的BLV感染奶牛在EBL亚临床期分离的外周血单核细胞亚群中BLV前病毒的分布。对5头BLV感染但临床正常的牛（前病毒载量> 100拷贝/1 × 105个细胞）进行表型表征，发现高百分比的CD 5 + IgM+细胞（但不是CD 5- IgM+ B细胞、CD 4 + T细胞或CD 8 +T细胞）。通过细胞分选或使用磁珠从五分之三的牛中纯化这些淋巴细胞亚群，并使用BLV-CoCoMo-qPCR估计BLV前病毒载量。所有动物中的CD 5 + IgM+ B细胞群比其他细胞群具有更高的BLV前病毒载量。感染CD 5- IgM+ B细胞、CD 4+细胞和CD 8 + T细胞的前病毒的拷贝数（每1 ml血液）分别为CD 5 + IgM+ B细胞中的1/34至1/4、1/22至1/3和1/31至1/3。此外，BLV前病毒仍然整合到CD 5 + IgM+ B细胞、CD 5- IgM+ B细胞、CD 4 + T细胞和CD 8 + T细胞的基因组DNA中，即使在前病毒载量<100拷贝/105细胞的BLV感染牛中也是如此。最近的研究结果表明，虽然CD 5 + IgM+ B细胞是BLV感染但临床正常牛的主要靶细胞类型，但CD 5- IgM+ B细胞、CD 4+细胞和CD 8 + T细胞的感染程度比以前认为的更大。
Bovine leukemia virus (BLV) is associated with enzootic bovine leukosis (EBL), which is the most common neoplastic disease of cattle. BLV infection may remain clinically silent at the aleukemic (AL) stage, cause persistent lymphocytosis (PL), or, more rarely, B cell lymphoma. BLV has been identified in B cells, CD2+ T cells, CD3+ T cells, CD4+ T cells, CD8+ T cells, γ/δ T cells, monocytes, and granulocytes in infected cattle that do not have tumors, although the most consistently infected cell is the CD5+ B cell. The mechanism by which BLV causes uncontrolled CD5+ B cell proliferation is unknown. Recently, we developed a new quantitative real-time polymerase chain reaction (PCR) method, BLV-CoCoMo-qPCR, which enabled us to demonstrate that the proviral load correlates not only with BLV infection, as assessed by syncytium formation, but also with BLV disease progression. The present study reports the distribution of BLV provirus in peripheral blood mononuclear cell subpopulations isolated from BLV-infected cows at the subclinical stage of EBL as examined by cell sorting and BLV-CoCoMo-qPCR. Phenotypic characterization of five BLV-infected but clinically normal cattle with a proviral load of > 100 copies per 1 × 105 cells identified a high percentage of CD5+ IgM+ cells (but not CD5- IgM+ B cells, CD4+ T cells, or CD8+T cells). These lymphocyte subpopulations were purified from three out of five cattle by cell sorting or using magnetic beads, and the BLV proviral load was estimated using BLV-CoCoMo-qPCR. The CD5+ IgM+ B cell population in all animals harbored a higher BLV proviral load than the other cell populations. The copy number of proviruses infecting CD5- IgM+ B cells, CD4+ cells, and CD8+ T cells (per 1 ml of blood) was 1/34 to 1/4, 1/22 to 1/3, and 1/31 to 1/3, respectively, compared with that in CD5+ IgM+ B cells. Moreover, the BLV provirus remained integrated into the genomic DNA of CD5+ IgM+ B cells, CD5- IgM+ B cells, CD4+ T cells, and CD8+ T cells, even in BLV-infected cattle with a proviral load of <100 copies per 105 cells. The results of the recent study showed that, although CD5+ IgM+ B cells were the main cell type targeted in BLV-infected but clinically normal cattle, CD5- IgM+ B cells, CD4+ cells, and CD8+ T cells were infected to a greater extent than previously thought.
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影响因子：
5.4
作者：
LAGARIAS, DM;RADKE, K
通讯作者：
RADKE, K
影响因子：
1.8
作者：
WILLIAMS, DL;BARTA, O;AMBORSKI, GF
通讯作者：
AMBORSKI, GF
DOI：
10.1073/pnas.79.8.2465
发表时间：
1982-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
KETTMANN, R;DESCHAMPS, J;MARBAIX, G
通讯作者：
MARBAIX, G
影响因子：
3.3
作者：
Jimba M;Takeshima SN;Matoba K;Endoh D;Aida Y
通讯作者：
Aida Y
影响因子：
5.4
作者：
Tajima, S;Ikawa, Y;Aida, Y
通讯作者：
Aida, Y