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[应用等电聚焦分析HLA I类分子。其在骨髓供体选择中的应用]

[Analysis of HLA class I molecules using isoelectric focusing. Its application in selection of bone marrow donors].

作者信息

Korínková P, Matĕjková E, Suttnar J, Vítek A

机构信息

Ustav hematologie a krevní transfuze, Praha.

出版信息

Cas Lek Cesk. 1997 May 7;136(9):279-81.

PMID:9264876
Abstract

BACKGROUND

Differences between HLA proteins class I are assessed by serological typing using HLA allo-antisera. This method suffices for the assessment of HLA signs in allogenic transplantations of bone marrow in genotypically identical siblings. However, it does not suffice in the selection of donors from the wider family of a related or not related donor from the register of voluntary bone marrow donors. In order to assess the polymorphism of the HLA-class I not detected by serological typing other more sensitive techniques are introduced. In the authors department one-dimensional isoelectric focusing was introduced as an auxiliary method for evaluation of identity of molecules in HLA-class I between bone marrow recipient and donor.

METHOD AND RESULTS

The authors introduced the method of one-dimensional isoelectric focusing which uses for detection of molecules of HLA-class I a polyclonal antibody against heavy chains of HLA molecules class I and a secondary antibody labelled by alkaline phosphatase. Forty-one pairs of bone marrow donors and recipient were examined. In six instances, i.e. in almost 15%, the authors detected by isoelectric focusing disagreement in HLA molecules class I. The authors present three interesting pairs of donors and recipients where isoelectric focusing helped with the selection of a suitable bone marrow donor.

CONCLUSIONS

The isoelectric focusing is at present another method which helps to reveal differences in HLA class I molecules. When methods of DNA analysis are introduced to recognize alleles of the HLA-I class, correlation with serologically and biochemically assessed variants wil be an essential guide in the evaluation of the final typing of HLA molecules class I.

摘要

背景

I类HLA蛋白之间的差异通过使用HLA同种异体抗血清的血清学分型来评估。该方法足以评估基因型相同的同胞之间骨髓同种异体移植中的HLA特征。然而,在从志愿骨髓供体登记册中选择来自相关或不相关供体的更广泛家族的供体时,该方法并不足够。为了评估血清学分型未检测到的I类HLA的多态性,引入了其他更敏感的技术。在作者所在科室,一维等电聚焦被引入作为评估骨髓受体和供体之间I类HLA分子同一性的辅助方法。

方法与结果

作者引入了一维等电聚焦方法,该方法使用针对I类HLA分子重链的多克隆抗体和碱性磷酸酶标记的二抗来检测I类HLA分子。对41对骨髓供体和受体进行了检查。在6例中,即近15%的情况下,作者通过等电聚焦检测到I类HLA分子存在不一致。作者展示了三对有趣的供体和受体,其中等电聚焦有助于选择合适的骨髓供体。

结论

等电聚焦目前是另一种有助于揭示I类HLA分子差异的方法。当引入DNA分析方法来识别I类HLA的等位基因时,与血清学和生化评估的变体的相关性将是评估I类HLA分子最终分型的重要指导。

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