American Society of Hematology

Figure 1.

$Figure 1. Peripheral blood findings and molecular evidence for loss of the telomeric region of chromosome 16p in a patient with acquired hemoglobin H and myelodysplastic syndrome. (A) Wright-Giemsa–stained peripheral blood smear demonstrates severe anisopoikilocytosis with hypochromic “ghost cells” almost completely devoid of hemoglobin (original magnification × 400). (B) Brilliant cresyl blue stain of peripheral blood reveals a “golf ball” cell (black arrow) with classical HbH inclusions (original magnification × 1000). Such cells represented only 0.11% of anucleate erythrocytes in the patient but were observed on multiple occasions. HbH-containing cells are not present in healthy persons. (C) Metaphase spread and 2 interphase nuclei from the patient's bone marrow hybridized with probe GG1 (green signal) reveals GG1 signal on the normal chromosome 16 only (original magnification, × 1000). Yellow arrowheads indicate the single GG1 signal in interphase, corresponding to the normal chromosome 16 short arm. Red arrow denotes the normal chromosome 16 homologue at metaphase, with paired GG1 signals on sister chromatids. (D) Southern blot of the 3′ hypervariable region in the alpha-globin cluster shows 2 bands for the 2 healthy controls (1 from each allele, corresponding to the normal diploid chromosome 16 short arm). In contrast, the second (upper) band from the patient's sample shows reduced probe hybridization. PB indicates DNA derived from unfractionated peripheral blood; Mono, DNA from the mononuclear cell fraction (predominantly lymphocytes); and Gran, the granulocyte-enriched fraction. The superior band is virtually absent from the patient's granulocyteenriched DNA, corresponding to the loss of an alpha-globin cluster in these cells. However, both bands are present in the mononuclear fraction, which is likely to be enriched for normal cells. In the patient's unfractionated peripheral blood, the superior band is less intense than the corresponding band in the mononuclear fragment, consistent with the expected admixture of clonal and nonclonal cells in unfractionated blood.$

Peripheral blood findings and molecular evidence for loss of the telomeric region of chromosome 16p in a patient with acquired hemoglobin H and myelodysplastic syndrome. (A) Wright-Giemsa–stained peripheral blood smear demonstrates severe anisopoikilocytosis with hypochromic “ghost cells” almost completely devoid of hemoglobin (original magnification × 400). (B) Brilliant cresyl blue stain of peripheral blood reveals a “golf ball” cell (black arrow) with classical HbH inclusions (original magnification × 1000). Such cells represented only 0.11% of anucleate erythrocytes in the patient but were observed on multiple occasions. HbH-containing cells are not present in healthy persons. (C) Metaphase spread and 2 interphase nuclei from the patient's bone marrow hybridized with probe GG1 (green signal) reveals GG1 signal on the normal chromosome 16 only (original magnification, × 1000). Yellow arrowheads indicate the single GG1 signal in interphase, corresponding to the normal chromosome 16 short arm. Red arrow denotes the normal chromosome 16 homologue at metaphase, with paired GG1 signals on sister chromatids. (D) Southern blot of the 3′ hypervariable region in the alpha-globin cluster shows 2 bands for the 2 healthy controls (1 from each allele, corresponding to the normal diploid chromosome 16 short arm). In contrast, the second (upper) band from the patient's sample shows reduced probe hybridization. PB indicates DNA derived from unfractionated peripheral blood; Mono, DNA from the mononuclear cell fraction (predominantly lymphocytes); and Gran, the granulocyte-enriched fraction. The superior band is virtually absent from the patient's granulocyteenriched DNA, corresponding to the loss of an alpha-globin cluster in these cells. However, both bands are present in the mononuclear fraction, which is likely to be enriched for normal cells. In the patient's unfractionated peripheral blood, the superior band is less intense than the corresponding band in the mononuclear fragment, consistent with the expected admixture of clonal and nonclonal cells in unfractionated blood.

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