Department of Clinical Neurosciences, Faculty of Medicine, University of Ostrava, Czech Republic.
Department of Biomedical Sciences, Faculty of Medicine, University of Ostrava, Czech Republic.
Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2023 Jun;167(2):121-130. doi: 10.5507/bp.2022.021. Epub 2022 May 4.
This study compared the results obtained by basic immunophenotyping of cerebrospinal fluid (CSF) cells by flow cytometry (FC) to the results of conventional cytology and evaluated the possibility of detailed analyses of CSF B-cell subpopulations.
Samples from 42 patients were examined by conventional cytology (native and/or pre-centrifuged CSF) and FC. The results from 15 patients without evidence of organic neurological disease were used to estimate reference ranges.
Pre-centrifugated CSF had significantly higher cell yield on the cytologic slide, but cell subpopulation percentages were altered; the percentage of lymphocytes was significantly higher and monocytes significantly lower compared to both native CSF slides and FC. The percentage of granulocytes was higher in FC compared to cytology. For leukocyte count, the following reference ranges were estimated for Fuchs-Rosenthal chamber (FR) counting and FC, respectively: leukocytes ≤4.7/μL and ≤2.5/μL, lymphocytes ≤4.1/μL and ≤1.8/μL, monocytes ≤1.2/μL and ≤0.9/μL, and granulocytes 0/μL and ≤0.2/μL. The following reference ranges were estimated for basic subpopulations: T-lymphocytes 84.1-100%, B lymphocytes 0.0-1.5%, NK cells 0.0-6.3%, NKT cells 0-9.5%, and CD3+CD4+/CD3+CD8+ 0.8-4.9. Using a volume of 1.2-2.4 mL, the number of B lymphocytes was too low (<20) in samples with ≤2.7 cells/μL in the FR.
Even normal CSF samples are amenable to basic mononuclear cell subpopulation analysis by FC. However, analysis of the B-cell subpopulations requires either a larger sample volume or selection of samples with ≥ 3 cells/μL.
本研究通过流式细胞术（FC）比较了脑脊液（CSF）细胞基本免疫表型的结果与传统细胞学的结果，并评估了详细分析 CSF B 细胞亚群的可能性。
对 42 例患者的标本进行了传统细胞学（原生和/或预离心 CSF）和 FC 检查。使用 15 例无器质性神经系统疾病证据的患者的结果来估计参考范围。
预离心 CSF 在细胞学载玻片上的细胞产量显著增加，但细胞亚群百分比发生了变化；与原生 CSF 载玻片和 FC 相比，淋巴细胞的百分比明显更高，单核细胞的百分比明显更低。FC 中的粒细胞百分比高于细胞学。对于白细胞计数，分别为 Fuchs-Rosenthal 室（FR）计数和 FC 估计了以下参考范围：白细胞≤4.7/μL 和≤2.5/μL，淋巴细胞≤4.1/μL 和≤1.8/μL，单核细胞≤1.2/μL 和≤0.9/μL，以及粒细胞 0/μL 和≤0.2/μL。为基本亚群估计了以下参考范围：T 淋巴细胞 84.1-100%，B 淋巴细胞 0.0-1.5%，NK 细胞 0.0-6.3%，NKT 细胞 0-9.5%，和 CD3+CD4+/CD3+CD8+0.8-4.9。使用 1.2-2.4mL 的体积，在 FR 中≤2.7 个细胞/μL 的样本中，B 淋巴细胞的数量过低（<20）。
即使是正常的 CSF 样本也可以通过 FC 进行基本单核细胞亚群分析。然而，B 细胞亚群的分析需要更大的样本量或选择≥3 个细胞/μL 的样本。
