Regéczy N, Görög G, Pálóczi K
Research Group of Hungarian Academy of Sciences, National Institute of Haematology and Immunology, Daróczi u. 24, H-1113 Budapest, Hungary.
Immunol Lett. 2001 May 1;77(1):47-54. doi: 10.1016/s0165-2478(01)00191-2.
In the process of developing a decision support system based on flow cytometric data for the diagnosis of immunodeficiency, assessment of lymphocyte subpopulations in human peripheral blood provides the key for further analysis. Samples from 273 'healthy' Hungarian subjects were measured between 1998 and 1999. Immunophenotypic data are compared here (unadjusted for gender) by different age groups: I 0-6 years (n=45); II 7-18 years (n=71); III 19-35 years (n=72); IV 36-55 years (n=48); and V 56-99 years (n=37). Two-color flow cytometric analysis was performed using the Becton Dickinson Simultest IMK Plus kit (CD45/CD14, isotype control, CD3/CD19, CD4/CD8, CD3/HLA-DR, CD3/CD16+56). All lymphocyte subpopulations were measured in all blood samples identically. The quality criteria involved at least 95% of total lymphocytes in the analysis gate, homogenous CD45+ lymphocyte population (minimum of 2000 events in the gate, CD45+ >95%). The frequency of B lymphocytes was the highest, significantly, in the youngest Hungarian subjects, but there were no significant changes with age comparing the data of other II-V age groups. On the other hand, some T subpopulations changed with aging; both CD4 and CD8 subsets varied over time including the elevation of the fraction of activated T cells as well as LGL-NK cells. Some of these changes were significant by statistical tests. Interpretation of flow cytometric data is time-consuming and requires human knowledge of an expert laboratory staff. To facilitate the diagnosis of immunodeficiency, a pilot study aiming at the development of a diagnostic algorithm has been initiated. Algorithm nodes compare the frequency of each lymphocyte subpopulations to the generated reference values. This knowledge-based system describes a short summary report as a result of the comparison, and points to some values requiring further human examination to reach a final conclusion. These reference values and the expert system appear to be a recommended basis for comparing and combining results from different laboratories.
在开发基于流式细胞术数据的免疫缺陷诊断决策支持系统的过程中，评估人类外周血中的淋巴细胞亚群为进一步分析提供了关键。1998年至1999年间对273名“健康”匈牙利受试者的样本进行了检测。在此比较不同年龄组（未按性别调整）的免疫表型数据：I组0 - 6岁（n = 45）；II组7 - 18岁（n = 71）；III组19 - 35岁（n = 72）；IV组36 - 55岁（n = 48）；V组56 - 99岁（n = 37）。使用Becton Dickinson Simultest IMK Plus试剂盒（CD45/CD14、同型对照、CD3/CD19、CD4/CD8、CD3/HLA - DR、CD3/CD16 + 56）进行双色流式细胞术分析。所有血液样本中所有淋巴细胞亚群的测量方法均相同。质量标准包括分析门中至少95%的总淋巴细胞、均匀的CD45 + 淋巴细胞群体（门中至少2000个事件，CD45 + > 95%）。B淋巴细胞的频率在最年轻的匈牙利受试者中显著最高，但比较其他II - V年龄组的数据时，随年龄没有显著变化。另一方面，一些T亚群随年龄变化；CD4和CD8亚群均随时间变化，包括活化T细胞以及大颗粒淋巴细胞 - 自然杀伤细胞（LGL - NK细胞）比例的升高。其中一些变化经统计检验具有显著性。流式细胞术数据的解读耗时且需要专业实验室人员的专业知识。为便于免疫缺陷的诊断，已启动一项旨在开发诊断算法的初步研究。算法节点将每个淋巴细胞亚群的频率与生成的参考值进行比较。这个基于知识的系统会根据比较结果生成一份简短的总结报告，并指出一些需要进一步人工检查以得出最终结论的值。这些参考值和专家系统似乎是比较和整合不同实验室结果的推荐基础。
