Dancey J T, Deubelbeiss K A, Harker L A, Finch C A
J Clin Invest. 1976 Sep;58(3):705-15. doi: 10.1172/JCI108517.
A method has been developed for measuring neutrophil cellularity in normal human bone marrow, in which the neutrophil-erythroid ratio was determined from marrow sections and marrow normoblasts were estimated by the erythron iron turnover. Neutrophil maturational categories, defined by morphologic criteria, were supported by autoradiographs of marrow flashed-labeled with 3H-thymidine. Correction for multiple counting error was empirically derived by counting serial sections through cells of each maturational category. The normal neutrophil-erythroid ratio in 13 normal human subjects was 1.5 +/- 0.07. The mean number of normoblasts in the same subjects was estimated to be 5.07 +/- 0.84 X 10(9) cells/kg. Total marrow neutrophils (X 10(9) cells/kg) were 7.70 +/- 1.20, the postmitotic pool (metamyelocytes, bands, and segmented forms) was 5.59 +/- 0.90 and the mitotic pool (promyelocytes + myelocytes) was 2.11 +/- 0.36. Marrow neutrophil ("total") production has been determined from the number of neutrophils comprising the postmitotic marrow pool divided by their transit time Transit time was derived from the appearance in circulating neutrophils of injected 3H-thymidine. The postmitotic pool comprised 5.59 +/- 0.90 X 10(9) neutrophils/kg, and the transit time was 6.60 +/- 0.03 days. From these data marrow neutrophil production was calculated to be 0.85 X 10(9) cells/kg per day. Effective production, measured as the turnover of circulating neutrophils labeled with 3H-thymidine, was 0.87 +/- 0.13 X 10(9) cells/kg per day. This value correlated well with the calculation of marrow neutrophil production. A larger turnover of 1.62 +/- 0.46 X 10(9) cells/kg per day was obtained when diisopropylfluorophosphate-32P was used to label circulating neutrophils. Studies using isologous cells doubly labeled with 3H-thymidine and diisopropylfluorophosphate-32P demonstrated a lower recovery and shorter t1/2 of the 32P label.
已开发出一种用于测量正常人骨髓中中性粒细胞细胞数的方法,该方法通过骨髓切片确定中性粒细胞与红细胞的比例,并通过红细胞铁周转率估算骨髓中幼红细胞。由形态学标准定义的中性粒细胞成熟类别,得到了用³H - 胸腺嘧啶闪烁标记骨髓的放射自显影片的支持。通过对每个成熟类别的细胞进行连续切片计数,凭经验得出了多重计数误差的校正值。13名正常受试者的正常中性粒细胞与红细胞比例为1.5±0.07。同一受试者中幼红细胞的平均数量估计为5.07±0.84×10⁹个细胞/千克。骨髓中性粒细胞总数(×10⁹个细胞/千克)为7.70±1.20,有丝分裂后池(晚幼粒细胞、杆状核细胞和分叶核细胞)为5.59±0.90,有丝分裂池(早幼粒细胞 + 中幼粒细胞)为2.11±0.36。骨髓中性粒细胞(“总数”)生成量是根据构成有丝分裂后骨髓池的中性粒细胞数量除以其转运时间来确定的。转运时间是从注射的³H - 胸腺嘧啶在循环中性粒细胞中的出现情况得出的。有丝分裂后池包含5.59±0.90×10⁹个中性粒细胞/千克,转运时间为6.60±0.03天。根据这些数据,计算出骨髓中性粒细胞生成量为0.85×10⁹个细胞/千克/天。以用³H - 胸腺嘧啶标记的循环中性粒细胞的周转率来衡量的有效生成量为0.87±0.13×10⁹个细胞/千克/天。该值与骨髓中性粒细胞生成量的计算结果相关性良好。当使用二异丙基氟磷酸 - ³²P标记循环中性粒细胞时,获得了更高的周转率,为1.62±0.46×10⁹个细胞/千克/天。使用³H - 胸腺嘧啶和二异丙基氟磷酸 - ³²P双重标记的同源细胞的研究表明,³²P标记的回收率较低且半衰期较短。