Han K, Kim Y, Lee J, Lim J, Lee K Y, Kang C S, Kim W I, Kim B K, Shim S I, Kim S M
Department of Clinical Pathology, Catholic University Medical College, Seoul, Korea.
Cytometry. 1999 Nov 1;37(3):178-83. doi: 10.1002/(sici)1097-0320(19991101)37:3<178::aid-cyto3>3.3.co;2-q.
Even modern automatic cell counters cannot count basophils precisely. Therefore, we need a rapid, accurate, precise, and easy method for counting basophils.
Using flow cytometry, basophils (CD22+/CD19-) and B cells (CD22+/CD19+) were counted. Within a large lymphocyte light scatter gate, % basophils (G%baso) and % B cells (G%B) were determined from the total count. Another method of analysis was to make two regions (R1 for basophils and R2 for B cells) and to determine in those the % basophils (R1%baso) and % B cells (R2%B) without gating. The flow cytometric basophil counts of the blood of 21 normal controls and 43 chronic myelogenous leukemia (CML) patients were compared with manual basophil count (Ma%baso) and basophil count by Coulter electronic cell counter (Hialeah, FL) (Auto%baso). CD22+/CD19- cells were sorted by a FACSCalibur (Becton Dickinson, San Jose, CA).
The G%baso of all samples was 4.66 +/- 5.35%, and R1%baso was 4.23 +/- 4.88%, and they were well-correlated (r = 0.996, P < 0.001). The G%B of all samples was 1.55 +/- 1.68%, and R2%B was 1.59 +/- 1.67%, and they were also well-correlated (r = 0.993, P < 0.001). Their correlation was better in normal controls than in CML. G%baso was well-correlated to Ma%baso (r = 0.827) and Auto%baso (r = 0.806), and R1%baso was well-correlated to Ma%baso (r = 0.831) but showed poor correlation to Auto%baso (r = 0.734). Auto%baso revealed the poorest correlation to Ma%baso (r = 0.692). The sorted CD22+/CD19- cells were all basophils (99.48 +/- 0.30%), and they revealed CD13, CD33, and dim CD45 expression, whereas CD3, CD14, CD16, and HLA-DR were not detected on them.
We discovered a specific marker combination to identify basophils (CD22+/CD19-), and we suggest that flow cytometric analysis using these markers is an easy, reliable, and accurate method of basophil counting.
即使是现代的自动细胞计数仪也无法精确计数嗜碱性粒细胞。因此，我们需要一种快速、准确、精确且简便的嗜碱性粒细胞计数方法。
使用流式细胞术对嗜碱性粒细胞（CD22+/CD19-）和B细胞（CD22+/CD19+）进行计数。在一个大淋巴细胞光散射门内，从总计数中确定嗜碱性粒细胞百分比（G%baso）和B细胞百分比（G%B）。另一种分析方法是划分两个区域（R1用于嗜碱性粒细胞，R2用于B细胞），并在不设门控的情况下确定其中的嗜碱性粒细胞百分比（R1%baso）和B细胞百分比（R2%B）。将21名正常对照者和43名慢性粒细胞白血病（CML）患者血液的流式细胞术嗜碱性粒细胞计数与手工嗜碱性粒细胞计数（Ma%baso）以及库尔特电子细胞计数仪（位于佛罗里达州希亚莱）的嗜碱性粒细胞计数（Auto%baso）进行比较。通过FACSCalibur（BD公司，加利福尼亚州圣何塞）对CD22+/CD19-细胞进行分选。
所有样本的G%baso为4.66±5.35%，R1%baso为4.23±4.88%，二者相关性良好（r = 0.996，P < 0.001）。所有样本的G%B为1.55±1.68%，R2%B为1.59±1.67%，二者相关性也良好（r = 0.993，P < 0.001）。它们在正常对照者中的相关性优于CML患者。G%baso与Ma%baso相关性良好（r = 0.827），与Auto%baso相关性良好（r = 0.806），R1%baso与Ma%baso相关性良好（r = 0.831），但与Auto%baso相关性较差（r = 0.734）。Auto%baso与Ma%baso的相关性最差（r = 0.692）。分选得到的CD22+/CD19-细胞均为嗜碱性粒细胞（99.48±0.30%），它们表达CD13、CD33和弱阳性CD45，而未检测到CD3、CD14、CD16和HLA-DR的表达。
我们发现了一种用于识别嗜碱性粒细胞的特异性标志物组合（CD22+/CD19-），并建议使用这些标志物的流式细胞术分析是一种简便、可靠且准确的嗜碱性粒细胞计数方法。
