Fetal Hemoglobin-Containing Cells Have the Same Mean Corpuscular Hemoglobin as Cells Without Fetal Hemoglobin: A Reciprocal Relationship Between Gamma- And Beta-Globin Gene Expression in Normal Subjects and in Those With High Fetal Hemoglobin Production
Fetal Hemoglobin-Containing Cells Have the Same Mean Corpuscular Hemoglobin as Cells Without Fetal Hemoglobin: A Reciprocal Relationship Between Gamma- And Beta-Globin Gene Expression in Normal Subjects and in Those With High Fetal Hemoglobin Production
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含有胎儿血红蛋白的细胞与不含胎儿血红蛋白的细胞具有相同的平均红细胞血红蛋白：正常受试者和胎儿血红蛋白产量高的受试者中 γ 和 β 珠蛋白基因表达之间的相互关系
DOI：
10.1182/blood.v69.4.1109.1109
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发表时间：
1987
期刊：
影响因子：
20.3
通讯作者：
S. Boyer
中科院分区：
文献类型：
--
作者：
G. Dover;S. Boyer
我们已经开发了一种方法，可以比较含胎儿血红蛋白（HbF）的红细胞（F细胞）的平均红细胞血红蛋白（MCH）与来自同一个体的非F细胞的MCH。为此，外周血红细胞及其内部内容物的悬浮液用亚氨二酯，二甲基-3，3 ′-二硫代双丙酰亚胺二盐酸盐（DTBP）固定。此后，通过用Triton X-100和异丙醇处理使固定的细胞对抗血清可渗透，与抗HbF的小鼠单克隆抗体（MoAb）反应，然后与荧光素缀合的抗小鼠IgG反应。在这种操作过程中没有明显的血红蛋白损失。因此，对来自不同受试者的红细胞进行了处理和显微镜检查，首先通过透射光，然后通过落射荧光。在选择的个体中，对于100个未固定（r = 0.96）和100个抗体处理的固定透化红细胞（r = 0.99），发现Coulter衍生MCH和415 nm透射光的平均吸光度之间存在直接相关性，以便提供20至35之间的Coulter MCH值范围。在所有受试者中，F细胞和非F细胞的显微镜下衍生MCH的比较在统计学上不可区分（P> 0.05）。这些比较包括正常个体（小于1% F细胞）、SS患者（7%至48% F细胞）、先天性贫血受试者（22%至65% F细胞）、HbF（HPFH）杂细胞遗传持久性个体（12%至21% F细胞）和β +地中海贫血杂合子（11%至31% F细胞）。我们的结论是，γ-和β-珠蛋白生产F细胞内的调节在一个互惠的方式之间的正常个体和个体之间的HbF生产升高。
We have developed methodology that allows comparison of the mean corpuscular hemoglobin (MCH) of fetal hemoglobin (HbF)-containing red cells (F cells) with the MCH of non-F cells from the same individual. To do this, suspensions of peripheral blood erythrocytes and their internal contents are fixed with an imidodiester, dimethyl-3,3′- dithiobispropionimidate dihydrochloride (DTBP). Thereafter fixed cells are made permeable to antisera by treatment with Triton X-100 and isopropanol, reacted with a mouse monoclonal antibody (MoAb) against HbF, and then with fluorescein-conjugated antimouse IgG. No appreciable hemoglobin is lost during such manipulation. Red cells from a diversity of subjects were thus treated and examined microscopically, first by transmitted light and then by epifluorescence. A direct correlation between Coulter-derived MCH and mean absorbance of 415 nm transmitted light was found for 100 unfixed (r = 0.96) and for 100 antibody-treated fixed-permeabilized red cells (r = 0.99) among individuals selected so as to provide a range of Coulter MCH values between 20 and 35. Comparisons of microscopically derived MCH of F cells and non-F cells were statistically nondistinguishable (P greater than 0.05) in all subjects. Such comparisons included normal individuals (less than 1% F cells), SS patients (7% to 48% F cells), subjects with congenital anemia (22% to 65% F cells), individuals with heterocellular hereditary persistence of HbF (HPFH) (12% to 21% F cells), and heterozygotes for beta + thalassemia (11% to 31% F cells). We conclude that gamma- and beta-globin production within F cells is regulated in a reciprocal fashion both among normal individuals and among individuals with elevated HbF production.