1.Expression change of IL-3 receptor system in all-trans retinoic acid induced differentiation of NB4 cells.
Yong WU ; Jing-Hui YANG ; Xian-Fang LI ; Xiao-Ying LIAO ; Hui-Fang HUANG ; Yuan-Zhong CHEN
Journal of Experimental Hematology 2010;18(6):1474-1478
Interleukin-3 receptor (IL-3R) is a heterodimeric membrane receptor. The α subunit is essential for ligand binding and confers ligand specificity to the receptor. The common beta chain (βc) subunit, which is shared by the granulocyte macrophage-colony stimulating factor (GM-CSF), IL-3 and IL-5 receptors, is required for high-affinity ligand binding and signal transduction, mediating growth and survival of hematopoietic progenitor cells and the production and activation of mature hematopoietic cells. In order to investigate the role of IL-3 receptor system (IL-3Rα, GM-CSFRα and hβc) in myeloid differentiation, the expression level of IL-3 receptor system gene in all-trans retinoic acid (ATRA)-induced NB4 cell differentiation was detected by quantitative real time RT-PCR. At the same time, DNA sequence change was analyzed by cDNA sequencing. The results showed that the expression level of IL-3Rα mRNA was obviously down-regulated in NB4 cells treated with ATRA for 24 hours, but during differentiation of ATRA induced NB4 cells, the expression level of IL-3Rα mRNA was gradually restored, while the expression levels of GM-CSFRα mRNA and hβc mRNA were gradually up-regulated. The sequence of IL-3Rα and GM-CSFRα gene did not change before and after NB4 cells differentiation, but the sequence of hβc gene changed when NB4 cells were treated with ATRA, the expression of hβc mRNA sequence before NB4 cell differentiation taken truncated mutation as dominant, as regards expression of hβc mRNA sequence after NB4 cell differentiation, the truncated mutation of hβc mRNA had restored to wild type. It is concluded that the IL-3 receptor abnormality exists in NB4 cells, over expression of IL-3Rα and truncated mutation of hβc may be involved in proliferation and differentiation block in NB4 cells.
Cell Differentiation
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drug effects
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Cell Line, Tumor
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Cytokine Receptor Common beta Subunit
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metabolism
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Humans
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Interleukin-3 Receptor alpha Subunit
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metabolism
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Signal Transduction
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Tretinoin
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pharmacology
2.Basophil Markers for Identification and Activation in the Indirect Basophil Activation Test by Flow Cytometry for Diagnosis of Autoimmune Urticaria.
Zehwan KIM ; Bong Seok CHOI ; Jong Kun KIM ; Dong Il WON
Annals of Laboratory Medicine 2016;36(1):28-35
BACKGROUND: The indirect basophil activation test using flow cytometry is a promising tool for autoimmune urticaria diagnosis. We aimed to identify better donor basophils (from atopic vs. non-atopic donors and interleukin-3 primed vs. unprimed basophils) and improve basophil identification and activation markers (eotaxin CC chemokine receptor-3 [CCR3] vs. CD123 and CD63 vs. CD203c). METHODS: Donor basophils were obtained from non-atopic and atopic group O donors. Positive control sera were artificially prepared to simulate autoimmune urticaria patients' sera. Patient sera were obtained from nine children with chronic urticaria. Assay sensitivity was compared among each variation by using positive control sera (n=21), applying cutoff values defined from negative control sera (n=20). RESULTS: For basophil identification, a combination of CCR3 and CD123 markers revealed a higher correlation with automated complete blood count (r=0.530) compared with that observed using CD123 (r=0.498) or CCR3 alone (r=0.195). Three activation markers on the atopic donor basophils attained 100% assay sensitivity: CD203c on unprimed basophils, CD63+CD203+ or CD63 alone on primed basophils; however, these markers on the non-atopic donor basophils attained lower assay sensitivity. CONCLUSIONS: For basophil identification markers, a combination of CD123 and CCR3 is recommended, while CD123 alone may be used as an alternative. Donor basophils should be obtained from an atopic donor. For basophil activation markers, either CD203c alone on unprimed basophils or CD203c and CD63 on primed basophils are recommended, while CD63 alone on primed basophils may be used as an alternative.
Autoimmune Diseases/blood/*diagnosis/immunology
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Basophils/*immunology/metabolism
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Biomarkers/blood
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Child
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Flow Cytometry
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Humans
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Interleukin-3 Receptor alpha Subunit/blood
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Male
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Receptors, CCR3/blood
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Urticaria/blood/*diagnosis/immunology
3.Basophil Activation Test for Chronic Urticaria.
Annals of Laboratory Medicine 2016;36(5):499-500
No abstract available.
Basophils/cytology/*metabolism
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Flow Cytometry
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HLA-DR Antigens/metabolism
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Humans
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Interleukin-3 Receptor alpha Subunit/metabolism
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Leukocyte Count
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Phosphoric Diester Hydrolases/metabolism
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Pyrophosphatases/metabolism
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Receptors, CCR3/metabolism
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Urticaria/*diagnosis/immunology/metabolism
4.A Rare Case of Acute Leukemic Presentation of Blastic Plasmacytoid Dendritic Cell Neoplasm without Cutaneous Lesions.
Shinae YU ; Min Jung KWON ; Kyungeun KIM ; Dong Hoe KOO ; Hee Yeon WOO ; Hyosoon PARK
Annals of Laboratory Medicine 2014;34(2):148-151
No abstract available.
Acute Disease
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Adult
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Antigens, CD4/metabolism
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Antigens, CD45/metabolism
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Antigens, CD56/metabolism
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Bone Marrow Cells/cytology
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Flow Cytometry
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Hematologic Neoplasms/*diagnosis/pathology
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Humans
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Interleukin-3 Receptor alpha Subunit/metabolism
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Lymph Nodes/metabolism/pathology
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Male
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Tomography, X-Ray Computed
5.CD34(+)CD19(+) cells with CD123 overexpression are a novel prognostic marker in Ph chromosome-positive acute lymphoblastic leukemia.
Yuan KONG ; Xiao-Jun HUANG ; Le HAO ; Ya-Zhen QIN ; Qian JIANG ; Hao JIANG ; Yan-Rong LIU
Journal of Experimental Hematology 2014;22(1):6-10
This study was aimed to investigate the characteristics of CD123 expression on CD34(+)CD19(+) cells and its prognostic significance as a novel MRD biomarker in Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL) patients. Consecutive newly diagnosed Ph(+)ALL patients (n = 49) in Peking University Institute of Hematology from January 2010 to April 2012 were prospectively enrolled in this study. At diagnosis and different time points during treatment, CD123 expression on CD34(+)CD19(+) cells was examined by multiparameter flow cytometry(MFC). More than 10 CD34(+)CD19(+) cells with CD123 overexpression in bone marrow samples after complete remission were defined as FCM positive (FCM(+)). The BCR-ABL1[STBZ] transcript was detected by real-time quantitative polymerase chain reaction (RQ-PCR) concurrently. The results showed that mean fluorescence intensity of CD123 on CD34(+)CD19(+) cells in newly diagnosed Ph(+)ALL and relapsed Ph(+)ALL patients was significantly higher than that of normal B-cell progenitors [8.52(3.71-32.35) vs 8.93(4.79-29.74) vs 1.31(0.21-1.75), P < 0.05]. In addition, ratio of the CD34(+)CD19(+) cells with CD123 overexpression in newly diagnosed Ph(+)ALL and relapsed Ph(+)ALL patients were significantly higher than that of normal B-cell progenitors [84.63% (55.07%-99.96%) vs 84.50% (57.68%-99.80%) vs 0.99% (0.45%- 1.83%), P < 0.05]. CD34(+)CD19(+) cells with CD123 overexpression were detected in all newly diagnosed and relapsed Ph(+)ALL patients. A good correlation was found between the MRD results of CD34(+)CD19(+) cells with CD123 overexpression detected by MFC and that detected by RQ-PCR (n = 360 pairs, Spearman r = 0.90, P < 0.0001). Among 13 cases relapsed during follow up, 11 cases of them were detected by FCM(+) at a median time of 60 (30-73) days before the recurrence. It is concluded that as a complementary to RQ-PCR, detection of the CD34(+)CD19(+) cells with CD123 overexpression by MFC promises to be an efficient tool for MRD assessment and risk stratification in human Ph(+)ALL.
Adolescent
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Adult
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B-Lymphocytes
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immunology
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metabolism
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Case-Control Studies
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Female
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Humans
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Interleukin-3 Receptor alpha Subunit
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metabolism
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Male
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Middle Aged
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Neoplasm Recurrence, Local
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Precursor Cell Lymphoblastic Leukemia-Lymphoma
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diagnosis
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immunology
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Prognosis
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Young Adult
6.Human BDCA2+CD123+CD56+ dendritic cells (DCs) related to blastic plasmacytoid dendritic cell neoplasm represent a unique myeloid DC subset.
Haisheng YU ; Peng ZHANG ; Xiangyun YIN ; Zhao YIN ; Quanxing SHI ; Ya CUI ; Guanyuan LIU ; Shouli WANG ; Pier Paolo PICCALUGA ; Taijiao JIANG ; Liguo ZHANG
Protein & Cell 2015;6(4):297-306
Dendritic cells (DCs) comprise two functionally distinct subsets: plasmacytoid DCs (pDCs) and myeloid DCs (mDCs). pDCs are specialized in rapid and massive secretion of type I interferon (IFN-I) in response to nucleic acids through Toll like receptor (TLR)-7 or TLR-9. In this report, we characterized a CD56(+) DC population that express typical pDC markers including CD123 and BDCA2 but produce much less IFN-I comparing with pDCs. In addition, CD56(+) DCs cluster together with mDCs but not pDCs by genome-wide transcriptional profiling. Accordingly, CD56(+) DCs functionally resemble mDCs by producing IL-12 upon TLR4 stimulation and priming naïve T cells without prior activation. These data suggest that the CD56(+) DCs represent a novel mDC subset mixed with some pDC features. A CD4(+)CD56(+) hematological malignancy was classified as blastic plasmacytoid dendritic cell neoplasm (BPDCN) due to its expression of characteristic molecules of pDCs. However, we demonstrated that BPDCN is closer to CD56(+) DCs than pDCs by global gene-expression profiling. Thus, we propose that the CD4(+)CD56(+) neoplasm may be a tumor counterpart of CD56(+) mDCs but not pDCs.
Biomarkers
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metabolism
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CD56 Antigen
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genetics
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immunology
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Cell Lineage
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genetics
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immunology
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Dendritic Cells
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immunology
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metabolism
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pathology
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Gene Expression
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Hematologic Neoplasms
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genetics
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immunology
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pathology
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Humans
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Immunophenotyping
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Interferon Type I
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biosynthesis
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metabolism
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Interleukin-12
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biosynthesis
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metabolism
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Interleukin-3 Receptor alpha Subunit
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genetics
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immunology
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Lectins, C-Type
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genetics
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immunology
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Membrane Glycoproteins
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genetics
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immunology
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Myeloid Cells
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immunology
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metabolism
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pathology
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Receptors, Immunologic
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genetics
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immunology
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Terminology as Topic
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Toll-Like Receptor 4
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genetics
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immunology
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Toll-Like Receptor 7
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genetics
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immunology
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Toll-Like Receptor 9
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genetics
;
immunology
7.Retrospective analysis of 4 cases of the so-called blastic NK-cell lymphoma, with reference to the 2008 WHO classification of tumours of haematopoietic and lymphoid tissues.
Yuan-yuan ZHENG ; Gang CHEN ; Xiao-ge ZHOU ; Yan JIN ; Jian-lan XIE ; Shu-hong ZHANG ; Yan-ning ZHANG
Chinese Journal of Pathology 2010;39(9):600-605
OBJECTIVETo study the clinical and pathologic features of 4 cases of the so-called blastic natural killer (NK)-cell lymphoma, with reference to the 2008 WHO classification of tumours of haematopoietic and lymphoid tissues.
METHODSThe clinical, pathologic and immunohistochemical findings (EliVision method) of 4 cases of blastic NK-cell lymphoma (previously diagnosed according to the 2001 WHO classification) were retrospectively analyzed and reclassified with a special reference to the 2008 WHO classification.
RESULTSThe 4 cases of hematologic malignancy studied were characterized by the presence of medium-sized blastic lymphoma cells, CD56 expression, and absence of lineage-specific B-cell, T-cell and myeloid cell markers. According to the 2001 WHO classification, they fell into the category of blastic NK-cell lymphoma. Three of the cases presented with primary cutaneous lesions and expression of CD56, CD4 and CD123. They are likely derived from the plasmacytoid dendritic cells rather than NK cells. They were then, according to the 2008 WHO classification, reclassified as the blastic plasmacytoid dendritic cell neoplasm. The remaining case showed lymph node involvement, positive for CD56 and CD4, negative for CD123, and not accompanied with the cutaneous lesions. This case was provisionally classified as a ambiguous lineage leukemia-NK cell lymphoblastic leukemia/lymphoma.
CONCLUSIONSThe so-called blastic NK-cell lymphomas in the 2001 WHO classification are rare and represent a heterogeneous group of lymphoproliferative disorders, with different clinical, pathologic and immunohistochemical features. It's suggested to have a precise category when applying the 2008 WHO classification to this kind of lesion.
Adult ; Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; CD56 Antigen ; metabolism ; Humans ; Interleukin-3 Receptor alpha Subunit ; metabolism ; Killer Cells, Natural ; pathology ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; classification ; metabolism ; pathology ; Retrospective Studies ; Skin Neoplasms ; classification ; metabolism ; pathology ; World Health Organization ; Young Adult
8.Expression of CD123 and CD114 on the bone marrow cells of patients with myelodysplastic syndrome.
Lan-zhu YUE ; Rong FU ; Hua-quan WANG ; Li-juan LI ; Hao-ran HU ; Lin FU ; Zong-hong SHAO
Chinese Medical Journal 2010;123(15):2034-2037
BACKGROUNDRecent studies have shown that interleukin-3 receptor alpha (CD123) is highly expressed on leukemia stem cells of patients with acute myeloid leukemia, and is correlated with tumor load and poor prognosis. The expression of CD123 may also be high in patients with myelodysplastic syndrome (MDS). In this study, the expression and clinical significance of CD123 and granulocyte colony stimulating factor (G-CSF) receptor (CD114) on the bone marrow cells of patients with MDS were investigated to explore the molecular marker of the malignant clone of MDS.
METHODSForty-two patients with MDS, who were diagnosed in the Hematological Department of General Hospital of Tianjin Medical University from 2008 to 2009, and twelve normal controls were enrolled in this study. Fluorescence activiated cell sorter (FACS) was used to measure the expression of CD123 on CD34(+)CD38(-) cells and CD114 on CD34(+) cells of the bone marrow of these patients and controls and the clinical significance was analyzed. The expression of CD114 on CD123(+)CD34(+)CD38(-) cells was further measured to explore the molecular marker of the malignant clone in MDS.
RESULTSMDS patients displayed significantly higher proportion of CD34(+)CD38(-)/CD34(+) ((14.03 +/- 5.27)%) than normal controls ((7.70 +/- 4.36)%, P < 0.05). The expression rate of CD123(+)CD34(+)CD38(-)/CD34(+)CD38(-) was significantly higher in MDS patients ((48.39 +/- 28.15)%) than that in normal controls ((8.75 +/- 11.71)%, P < 0.01). The expression level of CD123 was significantly correlated with the proportion of bone marrow blasts (r = 0.457, P < 0.05). The expression rate of CD114(+)CD34(+)/CD34(+) was lower in MDS patients ((33.05 +/- 21.71)%) than that in normal controls ((38.99 +/- 19.07)%) but was not statistically significant (P > 0.05). The expression of CD114 on CD123(+)CD34(+)CD38(-) cells ((34.82 +/- 29.58)%) was significantly lower than that on CD123(-)CD34(+)CD38(-) cells ((53.48 +/- 27.41)%) of MDS patients (P < 0.05).
CONCLUSIONSMDS patients displayed higher proportion of CD34(+)CD38(-)/CD34(+) than normal controls. CD123 was highly expressed in the bone marrow of the patients with MDS, significantly correlated with the proportion of bone marrow blasts, and thus might be the marker of MDS malignant clone. CD123(+)CD34(+)CD38(-) cells exhibited lower expression of G-CSF receptors, which might partly explain why MDS clone responds worse to G-CSF in vitro and in vivo.
ADP-ribosyl Cyclase 1 ; metabolism ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; metabolism ; Bone Marrow Cells ; metabolism ; Cells, Cultured ; Female ; Humans ; Interleukin-3 Receptor alpha Subunit ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; Receptors, Granulocyte Colony-Stimulating Factor ; metabolism ; Young Adult
9.Immunophenotyping of leukemic stem cells and chromosome karyotype characteristics in Uyghur leukemia pediatric patients.
Nuriding HAILIQIGULI ; Mei YAN
Chinese Journal of Oncology 2013;35(7):501-504
OBJECTIVETo study the immunophenotype and chromosome karyotype characteristics of leukemic stem cells (LSC) in Uyghur leukemia pediatric patients.
METHODSThe morphological features of LSC in culture in vitro was observed by flow cytometry. The immunophenotype was assessed by detective flow cytometry. The chromosome karyotype was analyzed by R-banding technique.
RESULTSThe LSC showed suspended floating colonies growing in the culture medium, and grew well and proliferated constantly in culture over 8 months. Among the 13 children with AML, there were 10 CD34(+)CD38(-)CD123(+) and CD33(+) cases, 10 CD44(+) cases, 10 CD96(+) cases, and 5 CD90(+) cases. Among the 13 children with B-ALL, there were 6 CD34(+)CD20(-)CD19(+) cases, 7 CD9(+) cases, and 5 CD123(+) cases. Among the 9 children with acute T lymphoblastic leukemia (T-ALL), there were 5 CD34(+)CD7(-) and CD90(+) cases, and 4 CD123(+) cases. Among the 13 cases of AML, 5 cases showed chromosome translocation t(15;17), one case chromosome translocation t(8;21), and 7 cases showed no chromosome karyotype abnormality. Among the 22 ALL cases, there were chromosome translocation t(12;21) in 1 case, t(9;22) in 3 case, hyperdiploid in 2 cases, and 16 cases without karyotype abnormalities. Twenty-nine children received induction remission therapy. Among them, 12 died, including 9 CD96(-)positive cases and 3 CD96(-)negative cases, with a statistically significant difference (P < 0.05).
CONCLUSIONSThe LSC of Uyghur leukemia pediatric patients in Xinjiang express CD9 and CD19 in ALL, and express CD123 and CD90 simultaneously in ALL and AML. The expression of CD96 is one of factors of poor prognosis.
Adolescent ; Antigens, CD ; metabolism ; Antigens, CD19 ; metabolism ; Child ; China ; ethnology ; Diploidy ; Humans ; Immunophenotyping ; Interleukin-3 Receptor alpha Subunit ; metabolism ; Karyotyping ; Leukemia, Myeloid, Acute ; drug therapy ; genetics ; immunology ; pathology ; Neoplastic Stem Cells ; immunology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; immunology ; pathology ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; immunology ; pathology ; Remission Induction ; Tetraspanin-29 ; metabolism ; Thy-1 Antigens ; metabolism ; Translocation, Genetic
10.Blastic plasmacytoid dendritic cell neoplasm: a clinicopathologic study.
Wei SANG ; Chao-fu WANG ; Yu-fan CHENG ; Xia LIU ; Xin-xia LI ; Abulajiang GULINAR ; Jun-zhi LI ; Wei ZHANG
Chinese Journal of Pathology 2012;41(5):326-330
OBJECTIVETo study the clinicopathologic features and differential diagnosis of blastic plasmacytoid dendritic cell neoplasm.
METHODSThe clinical, morphology and immunophenotypic features were analyzed in 3 cases of blastic plasmacytoid dendritic cell neoplasm, with review of literature.
RESULTSThe pathologic changes of these tumors accorded with that of blastic plasmacytoid dendritic cell neoplasm, and they also had new characteristics, including lineage other than T, B, myeloid and NK cells, and immunophenotypes of CD56(+) CD4(-) CD123(+) TdT(+) CD43(+) CD68(+) , CD56(+) CD4(+) CD123(-) TdT(+) CD43(+) CD68(-) and CD56(+) CD4(+) CD123(-/+) TdT(-) CD43(+) CD68(+) in the 3 cases, respectively. Bone marrow involvement was found 5 years later in case 1, and was then stable after chemotherapy; case 2 and case 3 were died 5 and 2 months after diagnosis, respectively.
CONCLUSIONBlastic plasmacytoid dendritic cell neoplasm is a heterogeneous group of lymphoproliferative disorders, with different clinical, morphologic and immunophenotypic features.
Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bleomycin ; therapeutic use ; CD56 Antigen ; metabolism ; Cyclophosphamide ; therapeutic use ; Dendritic Cells ; metabolism ; pathology ; Diagnosis, Differential ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Hematologic Neoplasms ; drug therapy ; metabolism ; pathology ; Humans ; Interleukin-3 Receptor alpha Subunit ; metabolism ; Leukemia, Myeloid ; metabolism ; pathology ; Lymphoma, Extranodal NK-T-Cell ; metabolism ; pathology ; Lymphoma, T-Cell, Peripheral ; metabolism ; pathology ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Prednisone ; therapeutic use ; Skin Neoplasms ; drug therapy ; metabolism ; pathology ; Treatment Outcome ; Vincristine ; therapeutic use