Title：HLA-specific B cells: I. A method for their detection, quantification, and isolation using HLA tetramers.Author：Alakulppi NS, Kyllonen LE, Partanen J, Salmela KT, Laine JT.
Source：Transplantation. 2007 Apr 15;83(7):982-8
Abstract：BACKGROUND: The development of highly sensitive and specific assays for detecting and characterizing HLA-specific antibodies has contributed to an appreciation of the extensive involvement of those antibodies in graft injury and dysfunction. However, understanding the regulatory processes of the humoral response to transplantation and the mechanisms underlying therapeutic agents and protocols for preventing and treating sensitization requires a way to study HLA-specific B cells. METHODS: Lymphocyte preparations enriched for B cells were stained with one or more of three different HLA tetramers. Tetramer-positive (tet+) B cells were enumerated and evaluated for an association of their frequencies with known sensitization. In some cases, tet+ B cells were isolated and placed in culture with supplements known to activate B cells in a nonspecific fashion. RESULTS: For all tetramers used, the frequencies of tet+ B cells were significantly higher (4.1%-5.5%) among sensitized patients than among nonsensitized patients (1.6%-3.2%, P<0.001). Binding of the tetramers occurred by the surface immunoglobulin antigen receptor with little or no binding to antibody captured in the Fc receptor. Cultured tet+ B cells produced antibodies specific for epitopes of the tetramer antigen. There appeared to be a certain amount of crossreactivity in the binding of tetramers. The frequency of CD27+ cells among tet+ B cells was higher, on average (34.4%-38.8%) than among all B cells (26.2%) whereas the frequencies of CD38 were comparable in the two groups. CONCLUSIONS: Staining with HLA tetramers provides a means for identifying, quantifying, and isolating HLA-specific B cells.
PMID: 17460571
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Title：HLA-specific B cells: I. A method for their detection, quantification, and isolation using HLA tetramers.
HLA特异性B细胞：一种借助于HLA四聚体对其进行的定性、定量和分离方法
Author：Alakulppi NS, Kyllonen LE, Partanen J, Salmela KT, Laine JT.
作者：Alakulppi NS, Kyllonen LE, Partanen J, Salmela KT, Laine JT.
Source：Transplantation. 2007 Apr 15;83(7):982-8
来源：移植杂志 期号:2007 年4月15日:83(7):982-8
Abstract：摘要
BACKGROUND: 研究现状
The development of highly sensitive and specific assays for detecting and characterizing HLA-specific antibodies has contributed to an appreciation of the extensive involvement of those antibodies in graft injury and dysfunction.
高敏感性及特异性的用以鉴定并将HLA特异性抗体特征化的分析技术的发展归功于针对广泛的涉及该类抗体在移植物损伤和功能障碍方面研究。
However, understanding the regulatory processes of the humoral response to transplantation and the mechanisms underlying therapeutic agents and protocols for preventing and treating sensitization requires a way to study HLA-specific B cells.
但是，要了解针对移植的体液反应的调节进程和隐在为预防和处置致敏作用而采取的多种治疗因素及方案之下的机制，需要一种用以研究HLA特异性B细胞的方法
METHODS:方法
Lymphocyte preparations enriched for B cells were stained with one or more of three different HLA tetramers. Tetramer-positive (tet+) B cells were enumerated and evaluated for an association of their frequencies with known sensitization. In some cases, tet+ B cells were isolated and placed in culture with supplements known to activate B cells in a nonspecific fashion.
为获取B细胞而对浓缩的淋巴细胞样品采用从三种不同的HLA 四聚物中选出的1种或更多进行染色。四聚物阳性(即tet+)的B细胞被借助其显现频率与已知的致敏作用的相互关联而计数和评估。在某些实验中，四聚物阳性B细胞被分离出来并被放置在含有可以非特异性方式激活B细胞的补体的培养基内。
RESULTS: For all tetramers used, the frequencies of tet+ B cells were significantly higher (4.1%-5.5%) among sensitized patients than among nonsensitized patients (1.6%-3.2%, P<0.001). Binding of the tetramers occurred by the surface immunoglobulin antigen receptor with little or no binding to antibody captured in the Fc receptor.
结果：对所应用的所有四聚物而言，四聚物阳性B细胞在致敏患者中的出现频率明显高(4.1%-5.5%)于非致敏患者(1.6%-3.2%, P<0.001)中的出现率。四聚物的结合是通过表面免疫球蛋白抗原受体借助于Fc受体与少量或者没有俘获过抗体的结合实现的。
Cultured tet+ B cells produced antibodies specific for epitopes of the tetramer antigen. There appeared to be a certain amount of crossreactivity in the binding of tetramers. The frequency of CD27+ cells among tet+ B cells was higher, on average (34.4%-38.8%) than among all B cells (26.2%) whereas the frequencies of CD38 were comparable in the two groups.
培养的四聚物阳性B细胞产生大量针对四聚物抗原的表位抗体。这提示在结合四聚物的反应中出现了一定数量的交叉反应。其出现频率在CD27+细胞中较四聚物阳性B细胞中为高，前者平均达34.4%-38.8%而在后者是26.2%。然而，CD38的表达率在两组细胞中均具有可比性。
CONCLUSIONS: Staining with HLA tetramers provides a means for identifying, quantifying, and isolating HLA-specific B cells.
结论：HLA四聚物染色法为HLA特异性B细胞的鉴别、定量、及分离提供了一种新的手段。
编译后：578字
HLA特异性B细胞：一种借助于HLA四聚体对其进行的定性、定量和分离方法
作者：Alakulppi NS, Kyllonen LE, Partanen J, Salmela KT, Laine JT.
来源：移植杂志 期号:2007 年4月15日:83(7):982-8
摘要
研究现状
高敏感性及特异性的用以鉴定并将HLA特异性抗体特征化的分析技术的发展归功于针对广泛的涉及该类抗体在移植物损伤和功能障碍方面研究。但是，要了解针对移植的体液反应的调节进程和隐在为预防和处置致敏作用而采取的多种治疗因素及方案之下的机制，需要一种用以研究HLA特异性B细胞的方法。
方法
为获取B细胞而对浓缩的淋巴细胞样品采用从三种不同的HLA 四聚物中选出的1种或更多进行染色。四聚物阳性(即tet+)的B细胞被借助其出现频率与已知的致敏作用的相互关联而计数和评估。在某些实验中，四聚物阳性B细胞被分离出来并被放置在含有可以非特异性方式激活B细胞的补体的培养基内。
结果：对所应用的所有四聚物而言，四聚物阳性B细胞在致敏患者中的出现频率明显高(4.1%-5.5%)于非致敏患者(1.6%-3.2%, P<0.001)中的出现率。四聚物的结合是通过表面免疫球蛋白抗原受体借助于Fc受体与少量或者没有俘获过抗体的结合实现的。培养的四聚物阳性B细胞产生大量针对四聚物抗原的表位抗体。这提示在结合四聚物的反应中出现了一定数量的交叉反应。其出现频率在CD27+细胞中较四聚物阳性B细胞中为高，前者平均达34.4%-38.8%而在后者是26.2%。然而，CD38的表达率在两组细胞中均具有可比性。
结论：HLA四聚物染色法为HLA特异性B细胞的鉴别、定量、及分离提供了一种新的手段。
文摘格式编译后如下：（593字）
移植杂志2007 年4月15日一期公布了一种借助于HLA四聚体对HLA特异性B细胞进行的定性、定量和分离方法：众所周知，高敏感性及特异性的用以鉴定并将HLA特异性抗体特征化的分析技术的发展归功于针对广泛的涉及该类抗体在移植物损伤和功能障碍方面研究。但是，要了解针对移植的体液反应的调节进程和隐在为预防和处置致敏作用而采取的多种治疗因素及方案之下的机制，需要一种用以研究HLA特异性B细胞的方法。该实验中，研究人员为获取B细胞而对浓缩的淋巴细胞样品采用从三种不同的HLA 四聚物中选出的1种或更多进行染色。四聚物阳性(即tet+)的B细胞被借助其出现频率与已知的致敏作用的相互关联而行计数和评估。在某些实验中，四聚物阳性B细胞被分离出来并被放置在含有可以非特异性方式激活B细胞的补体的培养基内。得到的结果是对所应用的所有四聚物而言，四聚物阳性B细胞在致敏患者中的出现频率明显高(4.1%-5.5%)于非致敏患者(1.6%-3.2%, P<0.001)中的出现率。四聚物的结合是通过表面免疫球蛋白抗原受体借助于Fc受体与少量或者没有俘获过抗体的结合实现的。培养的四聚物阳性B细胞产生大量针对四聚物抗原的表位抗体。这提示在结合四聚物的反应中出现了一定数量的交叉反应。其出现频率在CD27+细胞中较四聚物阳性B细胞中为高，前者平均达34.4%-38.8%而在后者是26.2%。然而，CD38的表达率在两组细胞中均具有可比性。研究者们得出的结论是HLA四聚物染色法为HLA特异性B细胞的鉴别、定量、及分离提供了一种新的手段。
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However, understanding the regulatory processes of the humoral response to transplantation and the mechanisms underlying therapeutic agents and protocols for preventing and treating sensitization requires a way to study HLA-specific B cells.
但是，要了解针对移植的体液反应的调节过程、潜在治疗因子的机制、预防和治疗致敏（患者）方案，需要研究HLA特异性B细胞。
Lymphocyte preparations enriched for B cells were stained with one or more of three different HLA tetramers. Tetramer-positive (tet+) B cells were enumerated and evaluated for an association of their frequencies with known sensitization. In some cases, tet+ B cells were isolated and placed in culture with supplements known to activate B cells in a nonspecific fashion.
富含B细胞的淋巴细胞标本用三种不同的HLA 四聚体中的1种或多种进行染色。对四聚体阳性(即tet+)的B细胞进行计数，评价其频率与已知的致敏作用的相互关联。在部分实验中，四聚物阳性B细胞被分离出来并放置在含有可以非特异性方式激活B细胞的补体的培养基内。
RESULTS: For all tetramers used, the frequencies of tet+ B cells were significantly higher (4.1%-5.5%) among sensitized patients than among nonsensitized patients (1.6%-3.2%, P<0.001). Binding of the tetramers occurred by the surface immunoglobulin antigen receptor with little or no binding to antibody captured in the Fc receptor.
结果：对所应用的所有四聚物而言，四聚物阳性B细胞在致敏患者中的出现频率明显高(4.1%-5.5%)于非致敏患者(1.6%-3.2%, P<0.001)中的出现率。四聚物的结合是通过表面免疫球蛋白抗原受体，通过Fc受体结合抗体的很少量甚至没有。
Cultured tet+ B cells produced antibodies specific for epitopes of the tetramer antigen. There appeared to be a certain amount of crossreactivity in the binding of tetramers. The frequency of CD27+ cells among tet+ B cells was higher, on average (34.4%-38.8%) than among all B cells (26.2%) whereas the frequencies of CD38 were comparable in the two groups.
培养的四聚体阳性B细胞产生针对四聚体抗原表位的特异性抗体。这提示在结合四聚体的反应中出现了一定数量的交叉反应。四聚体阳性B细胞中CD27+细胞出现频率高于其在总B细胞中的频率，前者平均达34.4%-38.8%而在后者是26.2%。然而，CD38的表达率在两组细胞中基本相当。
CONCLUSIONS: Staining with HLA tetramers provides a means for identifying, quantifying, and isolating HLA-specific B cells.
结论：HLA四聚体染色法为HLA特异性B细胞的鉴别、定量、及分离提供了一种新的手段。
