-
[show abstract]
[hide abstract]
ABSTRACT: Although quantitative and qualitative granulocyte defects have been described in myelodysplastic syndromes (MDS), the underlying molecular basis of granulocyte dysfunction in MDS is largely unknown. We recently found that FOS mRNA elevation under translation-inhibiting stimuli was significantly smaller in granulocytes from MDS patients than in healthy individuals. The aim of this study is to clarify the cause of the impaired FOS induction in MDS. We first examined the mechanisms of FOS mRNA elevation using granulocytes from healthy donors cultured with the translation inhibitor emetine. Emetine increased both transcription and mRNA stability of FOS. p38 MAPK inhibition abolished the emetine-induced increase of FOS transcription but did not affect FOS mRNA stabilization. The binding of an AU-rich element (ARE)-binding protein HuR to FOS mRNA containing an ARE in 3'UTR was increased by emetine, and the knockdown of HuR reduced the FOS mRNA stabilizing effect of emetine. We next compared the emetine-induced transcription and mRNA stabilization of FOS between MDS patients and healthy controls. Increased rates of FOS transcription by emetine were similar in MDS and controls. In the absence of emetine, FOS mRNA decayed to nearly 17% of initial levels in 45 min in both groups. In the presence of emetine, however, 76.7±19.8% of FOS mRNA remained after 45 min in healthy controls, versus 37.9±25.5% in MDS (P<0.01). To our knowledge, this is the first report demonstrating attenuation of stress-induced FOS mRNA stabilization in MDS granulocytes.
PLoS ONE 01/2013; 8(4):e61107. · 4.09 Impact Factor
-
Kayo Harada, Kazuhiko Ikeda,
Hayato Matsumoto,
Miki Furukawa,
Hiroshi Takahashi,
Hiroshi Ohkawara,
Hideyoshi Noji,
Kazuhiro Tasaki,
Masafumi Abe,
Kazuei Ogawa,
Yasuchika Takeishi
[show abstract]
[hide abstract]
ABSTRACT: Chronic lymphocytic leukemia (CLL) rarely exhibits an aggressive clinical course and its patients often have chromosomal deletions or additions. Furthermore, reciprocal translocations are barely observed in CLL. There have only been a few reports of CLL with t(1;6), and here we report the first Asian case of CLL with reciprocal translocation t(1;6). Since our case and previously reported CLL patients with t(1;6) consistently showed aggressive clinical course, t(1;6) may define a distinct type of CLL.
Experimental hematology & oncology. 09/2012; 1(1):28.
-
Huiyuan Hu,
Yayoi Shikama,
Tsutomu Shichishima, Kazuhiko Ikeda,
Kazuko Akutsu,
Tomoyuki Ono,
Hideo Kimura,
Kazuei Ogawa,
Hideyoshi Noji,
Yasuchika Takeishi,
Junko Kimura
[show abstract]
[hide abstract]
ABSTRACT: To investigate differentiation-dependent gene expression during granulopoiesis, we established a new method to isolate six sequential differentiation stages of neutrophil progenitors from bone marrow. Neutrophil progenitors were divided into three populations by density centrifugation, followed by depletion of other lineages, and further separated by fluorescence-activated cell sorting based on the expressions of CD34, CD11b, and CD16: CD34(+) fraction from a low-density population (F1), CD11b(-)/CD16(-) (F2), CD11b(+)/CD16(-) (F3), and CD11b(+)/CD16(low) (F4) fractions with intermediate density, and CD11b(+)/CD16(int) (F5) and CD11b(+)/CD16(high) (F6) fractions from a high-density population. To examine whether this fractionation was applicable to the study of in vivo gene expression profiles during granulopoiesis, we analyzed messenger RNA levels of AML-1 and CCAAT/enhancer binding protein (EBP)-ε and two target genes of C/EBP-ε, granulocyte-macrophage colony-stimulating factor receptor common β subunit and lactoferrin, in the six fractions and peripheral blood-derived neutrophils (F7). Expression of AML-1 and C/EBP-ε peaked at F1 and F4, respectively, followed by a gradual decrease. Although granulocyte-macrophage colony-stimulating factor receptor common β subunit messenger RNA levels remained low from F1 through F6 and elevated at F7, lactoferrin messenger RNA showed a drastic increase at F3 and dropped at F5. The difference in the expression profiles of the two C/EBP-ε target genes suggests the involvement of regulators other than C/EBP-ε in the induction of the two genes. The new fractionation method is able to provide new information on maturation-dependent gene expression during granulopoiesis.
Experimental hematology 04/2012; 40(8):675-81. · 3.11 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Philadelphia chromosome-negative myeloproliferative neoplasms (MPN), which include polycythemia vera, essential thrombocythemia, and primary myelofibrosis, are characterized by clonal proliferative hematopoiesis with increased blood cell count. Clonal expansion mechanisms in MPN and related disorders such as myelodysplastic syndromes (MDS) remain to be elucidated. Although mutations in the JAK2 gene lead to a proliferative hematopoiesis in majority of MPN and some MDS, the mutation alone does not cause a clonal expansion. In addition to JAK2 mutations, several genetic abnormalities, including TET2 and polycomb group genes involving epigenetic regulation have been reported in patients with MPN. Moreover, overexpression of HMGA2 due to removal of specific sites in its 3' untranslated region for regulatory let-7 micro RNAs may contribute to the proliferative hematopoiesis with conferring a growth advantage at the level of a hematopoietic stem cell in some cases with MPN.
Fukushima journal of medical science 01/2012; 58(2):91-100.
-
Alain M Ngoma, Kazuhiko Ikeda,
Yuko Hashimoto,
Kazuhiro Mochizuki,
Hiroshi Takahashi,
Hideki Sano,
Hayato Matsumoto,
Hideyoshi Noji,
Syunnichi Saito,
Atsushi Kikuta,
Kazuei Ogawa,
Mikio Ohtsuka,
Masafumi Abe,
Kenneth E Nollet,
Hitoshi Ohto
[show abstract]
[hide abstract]
ABSTRACT: To elucidate the correlation between regulatory T cells (Tregs) and acute graft-versus-host disease (aGVHD) or cytomegalovirus infection following allogeneic bone marrow transplantation (allo-BMT), we evaluated either CD4⁺CD25(high) or FOXP3⁺ Treg-enriched cells in peripheral blood (PB) from 20 patients who received allo-BMT, and in biopsies of skin with aGVHD. Proportions of CD4⁺CD25(high)FOXP3⁺ cells in total lymphocytes, but not other types of T cells, were lower in patients who eventually developed grades II-IV aGVHD (n = 13) than in others (n = 7, P < 0.001). Proportions of CD62L⁺ cells in CD4⁺CD25(high) cells at day +30 were lower (P < 0.01) in patients who eventually showed cytomegalovirus viremia (n = 6) than in others (n = 14). Incidence of aGVHD (P < 0.05) or cytomegalovirus viremia (P < 0.05) was higher in patients without these complications, but with lower proportions of PB CD4⁺CD25(high)FOXP3⁺ cells at day +30 (n = 8) than in others (n = 8). However, in skin with aGVHD (n = 5), there was marked or slightly increased infiltration of CD8⁺ cells (P < 0.001) or CD3⁺FOXP3⁺ cells (P < 0.05), respectively, when compared with control (n = 5), resulting in threefold higher ratio of CD8⁺/CD3⁺FOXP3⁺ cells in aGVHD relative to controls (P < 0.05). Thus, impaired reconstitution of Tregs may be associated with aGVHD and CMV infection. Moreover, imbalance of Tregs and CD8⁺ cells may play a role in aGVHD tissue.
International journal of hematology 12/2011; 95(1):86-94. · 1.17 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: An increasing number of medical centers can collect bone marrow, peripheral blood, or umbilical cord stem cells. Pathology laboratories should accommodate this trend, but investment in additional equipment may be impractical.
To compare CD34(+) cell counting results by using 2 widely available flow cytometry systems, with and without the use of a separate hematology analyzer (ie, single-platform versus dual-platform methodologies).
Whole blood and peripheral blood stem cell (PBSC) samples were analyzed from 13 healthy allogeneic PBSC donors and 46 autologous PBSC donors with various malignancies. The Cytomics FC500 (Beckman Coulter, Fullerton, California) was compared with the FACSCalibur (BD Biosciences, San Jose, California). Dual-platform CD34(+) cell counting incorporated data from a KX-21 hematology analyzer (Sysmex, Kobe, Japan).
Subtle differences in CD34(+) cell counting between 2 systems and 2 methods did not achieve statistical significance.
Different systems and methods for CD34(+) cell enumeration, properly validated, can support care for patients undergoing transplants and provide meaningful data for multicenter studies or meta-analyses.
Archives of pathology & laboratory medicine 07/2011; 135(7):909-14. · 2.58 Impact Factor
-
British Journal of Haematology 04/2011; 154(4):525-7. · 4.94 Impact Factor
-
Tomoaki Akagi,
Naoto Takahashi,
Kouhei Yamaguchi,
Kenichi Ishizawa,
Kazunori Murai,
Katsushi Tajima, Kazuhiko Ikeda,
Yoshihiro Kameoka,
Junnichi Kameoka,
Shigeki Ito,
Yuichi Kato,
Hideyoshi Noji,
Tsutomu Shichishima,
Jugoh Itoh,
Ryo Ichinohasama,
Hideo Harigae,
Yoji Ishida,
Kenichi Sawada
[show abstract]
[hide abstract]
ABSTRACT: To clarify the clinical outcome of peripheral T-cell lymphomas (PTCLs), we conducted a retrospective review comparing the outcomes of patients with PTCL (nodal peripheral T-cell lymphoma, unspecified, n=34 ; angioimmunoblastic T-cell lymphoma, n=12) to those with diffuse large B-cell lymphoma (DLBCL, n=48). All patients received CHOP-based chemotherapy without rituximab. PTCL patients presented at a more advanced clinical stage (91% vs. 65%, P<0.002) with a poorer performance status (26% vs. 17%, P<0.002) than DLBCL patients. The complete response rate among PTCL patients was significantly lower than among DLBCL patients (39% vs. 67%, P<0.008), as was the 3-year overall survival rate (26% vs. 50%, P=0.005), and Cox multivariate analysis revealed immunophenotype, performance status, and extranodal site involved to be significantly associated with shorter overall survival (P=0.045, P=0.007, and P=0.034, respectively). Our findings suggest that PTCL patients tend to have a poor prognosis associated with several initial risk factors. Moreover, the T-cell phenotype itself appears to have a significant impact on overall survival. Thus, standard CHOP chemotherapy may be inadequate for PTCLs, especially in patients with high-risk factors. The development of newly stratified therapies for the treatment of PTCLs would be highly desirable.
Journal of Clinical and Experimental Hematopathology 01/2011; 51(1):29-35.
-
Kazuei Ogawa, Kazuhiko Ikeda,
Miki Furukawa,
Kayo Harada-Shirado,
Yumiko Mashimo,
Hiroshi Takahashi,
Hayato Matsumoto,
Satoshi Kimura,
Akiko Shichishima-Nakamura,
Hiroshi Ohkawara,
Yuko Hashimoto,
Koichi Asahi,
Hideyoshi Noji,
Hitoshi Ohto,
Yasuchika Takeishi
[show abstract]
[hide abstract]
ABSTRACT: Renal amyloidosis is typically characterized by nephrotic syndrome, often with massive proteinuria and refractory peripheral edema. We report the case of a patient with renal amyloidosis associated with nephrotic syndrome who maintained remission for 6 years after undergoing high-dose chemotherapy followed by autologous peripheral blood stem-cell transplantation (auto-PBSCT). The patient was a man aged in his 50s who had developed nephrotic syndrome. Bone marrow aspiration and kidney biopsy determined that the cause of the nephrotic syndrome was renal amyloidosis due to multiple myeloma, and the patient was admitted to our department in July 2003. After one course of chemotherapy, auto-PBSCT was performed in March 2004. Following transplantation, serum M-protein was no longer detectable from March 2005, and the patient achieved complete hematological remission. Subsequently, proteinuria decreased, serum albumin levels normalized, and nephrotic syndrome improved. As of 6 years after transplantation, in March 2010, the patient remained in remission, meaning that auto-PBSCT proved extremely effective as a treatment for renal amyloidosis in this case.
Fukushima journal of medical science 12/2010; 56(2):151-6.
-
Kazuei Ogawa,
Hideyoshi Noji,
Miki Furukawa,
Kayo Harada-Shirado,
Yumiko Mashimo,
Hiroshi Takahashi,
Hayato Matsumoto,
Satoshi Kimura,
Akiko Shichishima-Nakamura,
Hiroshi Ohkawara, Kazuhiko Ikeda,
Hitoshi Ohto,
Yasuchika Takeishi
[show abstract]
[hide abstract]
ABSTRACT: From 1996 to the end of 2009, a total of 114 cases of hematopoietic stem cell transplantation were performed in the Department of Hematology, Fukushima Medical University. We report here a general overview of our results. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) was performed in 37 cases of acute leukemia, 10 of myelodysplastic syndrome, 5 of aplastic anemia, and 5 others. The 5-year survival rate with allo-HSCT was 51.1%. Autologous hematopoietic stem cell transplantation (auto-HSCT) was performed in 34 cases of malignant lymphoma, 15 of multiple myeloma, and 8 others. The 5-year patient survival rate was 75.2% with malignant lymphoma and 46.7% with multiple myeloma. These results are comparable to those from a nationwide survey in Japan, confirming that our hospital has attained a creditable level as a transplantation center.
Fukushima journal of medical science 12/2010; 56(2):107-14.
-
[show abstract]
[hide abstract]
ABSTRACT: A 27-year-old woman exhibited progressive pancytopenia during cyclophosphamide pulse therapy for lupus nephritis and low-dose methotrexate therapy for severe arthralgia. Bone marrow aspiration revealed highly abnormal cell morphology, indicating therapy-related myelodysplastic syndrome. Pancytopenia and bone marrow cell morphology improved 3 months after discontinuation of cyclophosphamide. It is necessary to promptly examine bone marrow cell morphology and chromosomal aberration in cases with connective tissue diseases complicated by sudden cytopenia during immunosuppressive therapy with chemotherapeutic agents.
Fukushima journal of medical science 12/2010; 56(2):121-7.
-
Tsutomu Shichishima, Kazuhiko Ikeda,
Naoto Takahashi,
Junichi Kameoka,
Katsushi Tajima,
Kazunori Murai,
Yoshiko Tamai,
Akiko Shichishima-Nakamura,
Kazuko Akutsu,
Hideyoshi Noji,
Masatoshi Okamoto,
Hideo Kimura,
Hideo Harigae,
Takashi Oyamada,
Toyomi Kamesaki,
Yasuchika Takeishi,
Kenichi Sawada
[show abstract]
[hide abstract]
ABSTRACT: To clarify whether measurement of serum haptoglobin (Hp) has impact on understanding pathophysiology in bone marrow failure (BMF) syndromes, we investigated concentrations of serum Hp by nephelometric procedure in 156 Japanese patients with BMF, including 54 aplastic anemia (AA), 50 paroxysmal nocturnal hemoglobinuria (PNH), and 52 myelodysplastic syndromes (MDS) patients. The frequencies with low concentrations of serum Hp (<42 mg/dL) in PNH patients (98.0%) were significantly higher than those in AA (27.8%; P < 0.0001) and MDS (38.5%; P < 0.0001) patients. In AA patients, white blood cell (WBC), absolute neutrophil, and platelet counts were significantly decreased in the group (n = 15) with low concentrations of serum Hp than in that (n = 39) with normal concentrations of it, and WBC counts were positively correlated with concentrations of serum Hp, suggesting that WBC counts may affect the concentrations. In MDS patients, hemoglobin concentrations and serum iron were significantly decreased and increased, respectively, in the group (n = 20) with low concentrations of serum Hp than in that (n = 32) with normal concentrations of it, and the values of serum iron were inversely correlated with concentrations of serum Hp, suggesting that ineffective erythropoiesis may affect the concentrations. Several AA and MDS patients with low concentrations of serum Hp had Coombs-negative autoimmune hemolytic anemia determined by immunoradiometric assay. In conclusion, several factors in conjunction with pathophysiology contribute to decrease of serum Hp in BMF.
International journal of hematology 04/2010; 91(4):602-10. · 1.17 Impact Factor
-
Nihon Naika Gakkai Zasshi 02/2009; 98(1):141-3.
-
[show abstract]
[hide abstract]
ABSTRACT: Although automated programs have been increasingly used to collect peripheral blood (PB) progenitor cells (PBPCs), differences among them remain unclear. The automated programs of Amicus (Baxter Healthcare) and Spectra (software Version 6.1, Gambro BCT) apheresis machines were compared in a crossover study.
The patients for autologous and donors for allogeneic PBPC transplantation mobilized with granulocyte-colony-stimulating factor were randomly assigned into two groups. PBPCs were collected by the Amicus on the first day and the Spectra on the second of 2 consecutive days in Group I, and the reverse order was used in Group II. Of 39 patients or donors enrolled, 17 reached their collection goal with only one procedure and did not participate in the paired study. Thus, 44 paired procedures of the remaining 22 subjects were evaluated.
The product yields of white blood cells (WBCs; p < 0.005), mononuclear cells (MNCs; p < 0.02), and CD34+ PBPCs (p < 0.0002) from patients or donors were higher in the Amicus collections than those in the Spectra collections. The collection efficiencies of WBCs (p < 0.03), MNCs (p < 0.02), and CD34+ PBPCs (p < 0.03) were higher in the Amicus collections. The numbers of contaminating platelets (PLTs) in the Amicus collections were lower than those in the Spectra collections (p < 0.05) with a greater decrease in PB PLT counts after apheresis with the Spectra (p < 0.01). The Amicus had a longer running time than the Spectra for processing similar volumes (p < 0.005).
The automated program of the Amicus may be better than that of the Spectra for collecting MNCs and CD34+ PBPCs and avoiding apheresis-induced thrombocytopenia.
Transfusion 08/2007; 47(7):1234-40. · 3.22 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To clarify an expansion mechanism of a paroxysmal nocturnal hemoglobinuria (PNH) clone with the Wilms' tumor gene (WT1).
In PNH patients with the HLA-A*2402 allele, frequencies of peripheral blood (PB) WT1 peptide-specific and HLA-A*2402-restricted CD8+ cells and WT1 peptide-stimulated interferon-gamma-producing mononuclear cells (MNCs), cytotoxicity of WT1 peptide-specific and HLA-A*2402-restricted cytotoxic T lymphocyte (CTL) clone (TAK-1) cells on bone marrow (BM) MNCs, and after co-incubation with TAK-1 cells, changes in colony-forming unit granulocyte-macrophage colony formation of CD34+ cells and in CD59 expression in viable CD34+ cells were investigated.
The frequencies of PB WT1 peptide-specific and HLA-A*2402-restricted CD8+ cells (p < 0.005) and WT1 peptide-stimulated interferon-gamma-producing MNCs (p < 0.02) were significantly higher in 5 PNH patients than 8 healthy volunteers (HV). In 5 PNH patients or 3 HV, TAK-1 cells significantly killed BMMNCs and suppressed colony formations of CD34+CD59+ and/or CD34+CD59- cells in the absence and presence of a WT1 peptide or only in the presence of the peptide, respectively, in an HLA-restricted manner. After co-incubation with TAK-1 cells, reduction rates of colony formation of CD34+CD59- cells were significantly less than those of CD34+CD59+ cells in 5 PNH patients (p < 0.002) and proportions of viable CD34+CD59- cells from 5 PNH patients significantly increased in the absence (p < 0.01) and presence (p < 0.01) of a WT1 peptide in an HLA-restricted manner.
WT1 peptide-specific and HLA-restricted CTLs may play an important role in expansion of a PNH clone during immunologic selection and/or in the occurrence of BM failure via interferon-gamma in PNH.
Experimental Hematology 05/2007; 35(4):618-26. · 2.90 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The frequencies of the HLA-B*4002 and HLA-A*0206 alleles in patients with aplastic anemia (AA) (n=32; 21.9%) and paroxysmal nocturanl hemoglobinuria (PNH) (n=24; 22.9%), respectively, were significantly different from those in controls (n=371; 8.6%, p<0.002 and 7.7%, p<0.001, respectively), suggesting that each specific allele in AA or PNH may be related to the immunologic pathophysiology of these disorders.
Haematologica 07/2006; 91(6):856-7. · 6.42 Impact Factor
-
Kazuhiko Ikeda,
Tsutomu Shichishima,
Takanori Teshima,
Kazuei Ogawa,
Akiko Nakamura-Shichishima,
Hiroko Tajima,
Hideyoshi Noji,
Yuko Hashimoto,
Kunihiko Takeyama,
Toshiyuki Ishibashi,
Hitoshi Ohto,
Masafumi Abe,
Yukio Maruyama
[show abstract]
[hide abstract]
ABSTRACT: Host-derived Langerhans cells (LCs) are crucial antigen-presenting cells that cause graft-vs.-host disease after allogeneic haematopoietic stem cell transplantation (HSCT). However, chimaerism of LCs after allogeneic HSCT is largely unknown in humans. We here report a case that developed dermatopathic lymphadenitis accompanied by an accumulation of donor-derived LCs in the second month after allogeneic HSCT with reduced-intensity conditioning. This is the first case to show that donor LCs have the ability to migrate into draining lymph nodes and replace host LCs early after HSCT in humans.
European Journal Of Haematology 04/2006; 76(3):261-4. · 2.61 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: A patient who had myelodysplastic syndrome (MDS) and dilated cardiomyopathy (DCM) had a transient improvement of cardiac function after peripheral blood stem cell transplantation (PBSCT). When he was admitted to hospital for PBSCT, a chest X ray showed cardiomegaly, and Tc-99m quantitative gated single photon emission computed tomography (QGS) showed increases in left ventricular (LV) volumes and a decrease in LV ejection fraction (LVEF). A coronary angiogram showed no evidence of coronary artery disease. Left ventriculography showed similar findings as QGS, and the findings from a myocardial biopsy were compatible with DCM. Three months after a successful allo-PBSCT with his brother as the donor, the cardiomegaly had been attenuated, the LV volumes decreased and LVEF increased on the QGS images. However, 10 months later, his cardiac function had deteriorated. The changes in cardiac function did not correlate with the hematological changes, such as the hemoglobin level.
Circulation Journal 11/2004; 68(10):958-60. · 3.77 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Efficient collection of progenitor cells is essential for PBPC transplantation. Two apheresis machines (Amicus, Baxter Healthcare; and Spectra, Gambro BCT, software version 4.7) were compared prospectively by a crossover trial.
Apheresis collections were performed for two consecutive days on patients for autologous and donors for allogeneic PBPC transplantation. The patients and donors, receiving a G-CSF, were randomized into two groups. In Group I, PBPCs were collected by the Amicus on the 1st day and the Spectra on the 2nd day, and the reverse order was used with Group II. A total of 60 apheresis procedures of 30 (16 in Group I and 14 in Group II) among 40 patients and donors enrolled were performed and evaluated.
The nucleated cell counts, MNC counts, CD34+ PBPC counts, and amounts of CFU-GM collected per procedure were similar with the Amicus and the Spectra. On the other hand, the decrease of peripheral blood platelet counts of patients and donors was more prominent from using Spectra than Amicus (p < 0.0001). Components collected by the Amicus had fewer platelets than those collected by the Spectra (p < 0.0001). The efficiencies of collecting nucleated cells, MNCs, and CD34+ PBPCs were not different between the machines (p > 0.05). However, the efficiency of collecting platelets was significantly higher with Spectra than with Amicus (p < 0.0001). The Amicus took longer than the Spectra to process the same volume (p < 0.05).
Amicus is superior to Spectra in avoiding apheresis-induced thrombocytopenia caused by platelets contaminating the collected samples. Therefore, the Amicus is useful for patients with thrombocytopenia or with a less-than-normal platelet count.
Transfusion 07/2003; 43(6):814-9. · 3.22 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To define the phosphatidylinositol glycan-class A (PIG-A) gene abnormality in precursor cells and the changes of expression of glycosylphosphatidylinositol-anchored protein and contribution of paroxysmal nocturnal hemoglobinuria (PNH) clones with PIG-A gene abnormalities among various cell lineages during differentiation and maturation, we investigated CD59 expression on bone marrow CD34(+) cells and peripheral granulocytes from 3 patients with PNH and the PIG-A gene abnormalities in the CD59(-), CD59(+/-), and CD59(+) populations by nucleotide sequence analyses. We also performed clonogeneic assays of CD34(+)CD59(+) and CD34(+)CD59(-) cells from 2 of the patients and examined the PIG-A gene abnormalities in the cultured cells. In case 1, the CD34(+) cells and granulocytes consisted of CD59(-) and CD59(+) populations and CD59(-), CD59(+/-), and CD59(+) populations, respectively. Sequence analyses indicated that mutation 1-2 was in the CD59(+/-) granulocyte population (20 of 20) and the CD34(+)CD59(-) population (2 of 38). In cases 2 and 3, the CD34(+) cells and granulocytes consisted of CD59(+) and CD59(-) cells. Sequence analyses in case 3 showed that mutation 3-2 was not in CD34(+)CD59(-) cells and was present in the CD59(-) granulocyte population. However, PIG-A gene analysis of cultured CD34(+)CD59(-) cells showed that they had the mutation. This analysis also revealed that there were some other mutations, which were not found in CD34(+)CD59(-) cells and CD59(-) or CD59(+/-) granulocytes in vivo, and that sometimes they were distributed specifically among different cell lineages. In conclusion, our findings suggest that PNH clones might contribute qualitatively and quantitatively differentially to specific blood cell lineages during differentiation and maturation of hematopoietic stem cells.
Blood 12/2002; 100(10):3812-8. · 9.90 Impact Factor
