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Imbalance of Th17 and Treg cells in peripheral blood from multiple myeloma patients.

Jing-jing LI ; Qian NIU ; Di-jiao TANG ; Neng-gang JIANG ; Yong-mei JIN ; Jun SU ; Yong-qian JIA

Chinese Journal of Hematology.2013;34(11):936-940. doi:10.3760/cma.j.issn.0253-2727.2013.11.006

OBJECTIVETo investigate the ratio of Th17 cells and CD4⁺CD25⁺Foxp3⁺ regulatory T (Treg) cells in peripheral blood from patients with multiple myeloma (MM) and explore its pathological effects.

METHODS70 MM patients were divided into three groups: newly diagnosed group (n=30), plateau stage group (n=23) and relapsed/refractory group (n=17). The controls consisted of 20 healthy donors. The frequencies of Th17 and Treg cells were detected by flow cytometry.

RESULTSCompared with controls [(0.72±0.33)%] and plateau stage group [(0.74±0.29)%], frequencies of Th17 cells were higher in newly diagnosed group [(1.62±0.65)%] and relapsed/refractory group [(1.45±0.51)%], respectively (P<0.05). Compared with controls [(2.33±0.90)%] and plateau stage group [(1.69±0.70)%], frequencies of Treg cells were significantly lower in newly diagnosed group [(0.55±0.23)%] and relapsed/refractory group [(0.82±0.54)%], respectively (P<0.05). The ratios of Th17/Treg in newly diagnosed group and relapsed/refractory group were higher than those in controls (P<0.05). There were no differences of the frequencies of CD3⁺CD4⁺ T cells and Th17 cells between plateau stage group and controls. The frequencies of Treg cells were significantly lower in plateau stage group than that in controls (P<0.05), and the ratio of Th17/Treg was significantly higher in plateau stage group than that in controls (P<0.05).

CONCLUSIONThe remarkable abnormality of T cells subsets was reduction of CD4⁺ T cells in MM. Higher frequency of Th17 and lower ratio of Treg could lead to imbalance of Th17/Treg, which may play a critical role in the pathogenesis of MM.

Adult ; Aged ; Female ; Flow Cytometry ; Humans ; Lymphocyte Count ; Male ; Middle Aged ; Multiple Myeloma ; immunology ; pathology ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology

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Apoptosis and its mechanisms of spleen CD4⁺ CD25⁺ regulatory T cells in severe aplastic anemia mouse model.

Wen-ping LU ; Zeng-hua LIN ; Hong LIU ; Wei LU ; Hai-yan LIU ; Xu-li WANG

Chinese Journal of Hematology.2013;34(11):931-935. doi:10.3760/cma.j.issn.0253-2727.2013.11.005

OBJECTIVETo explore the apoptosis and its mechanisms of spleen CD4⁺ CD25⁺ regulatory T (Treg) cells in severe aplastic anemia (SAA) mouse model induced by interferon (IFN-γ) in combination with busulphan (BU).

METHODSThe BALB/c female mice SAA model was induced by intraperitoneal injection with IFN-γ and intragastric administration with BU (combined group, n=16), with BU group (n=16), IFN-γ group (n=16) and normal group (n=16) as controls. Spleen Treg cells were purified by using of immunomagnetic beads. Apoptosis was detected by flow cytometry. AKt and TGF-β expression was measured by Western blot.

RESULTSApoptosis of spleen Treg cells in combined group [(33.21±0.65)%] was significantly higher than that in BU group [(21.58±0.64)%], IFN-γ group [(17.62±1.05)%], and control[(27.38±1.89)%] (P<0.05). A significantly lower expression of AKt and TGF-β protein was also seen in combined group (0.30±0.05 and 0.17±0.05), as compared to the other three groups (P<0.05).

CONCLUSIONExcessive apoptosis of Treg cells was found in SAA mouse model, which may be a cause of Treg cells decrease in patients with AA. The down-regulated expression of AKt and TGF-β could play a role in increased apoptosis of Treg cells. Our data may provide a new treatment strategy in AA.

Anemia, Aplastic ; prevention & control ; Animals ; Apoptosis ; Disease Models, Animal ; Female ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; T-Lymphocytes, Regulatory ; cytology

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Study of RON mediated invasion of Raji cell line and drug-target effects.

Bi-cui ZHAN ; Yue-han DONG ; Jian FAN ; Hang-ping YAO ; Jie JIN ; Xiang-min TONG

Chinese Journal of Hematology.2013;34(11):926-930. doi:10.3760/cma.j.issn.0253-2727.2013.11.004

OBJECTIVETo study the proto-oncogene RON mediated aggression of Raji cells and the inhibitory effects by monoclonal antibody Zt/f2 (2f2).

METHODSThe effects of RON ligand macrophage stimulating protein (MSP) (2.0 nmol/L) and inhibitory Zt/f2 (2F2) (2.0 nmol/L) antibody on proliferation of RON positive Raji cells after treatment for 24 and72 hours were detected by MTT method, colony formation units (CFU) of Raji cells by methylcellulose semi solid culture, Raji cells apoptosis and cell cycle analysis by AnnexinV/PI double staining, expression of RON, apoptosis-related proteins, and cyclins by Western blot.

RESULTS(1)Compared with the cell viability (1.0) and counts of CFU (103.6±7.0) in control group, Raji cells after MSP treatment had better viability (1.35±0.20) and CFU counts (133.7±10.4) (P<0.05), but worse viability (0.68±0.11) and CFU counts (66.3±6.1) after Zt/f2 (2F2) treatment (P<0.05). (2)Percentage of Raji cells apoptosis after Zt/f2 (2F2) antibody treatment (12.16±2.33)% was significantly increased than the control (2.89±1.03)% (P<0.05). The percentage of Raji cells arrested in G0/G1 phase was increased after Zt/f2 (2F2) antibody treatment as compared to the control [ (54.96 ±3.70)% vs (39.10±2.30)%, (P<0.05) ]. (3) High-level of RON phosphorylation and β-catenin expression activated by MSP could be inhibited significantly by Zt/f2 (2F2), which also up-regulated the expression of caspase-3, caspase-8, caspase-9 and PARP and down-regulated anti-apoptotic MCL-1 gene and inhibitor of apoptosis protein XIAP expression, accompanied with G1 phase protein changes accordingly.

CONCLUSIONMSP could aggravate Raji cells proliferation. Inversely, Zt/f2 (2F2) could inhibit proliferation and induce apoptosis by inhibition of RON phosphorylation and up-regulation of apoptosis related proteins.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Proto-Oncogenes ; Receptor Protein-Tyrosine Kinases ; metabolism

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Prophylactic G-CSF mobilized donor lymphocytes infusion after non-myeloablative stem cell transplantation prevents relapse in patients with high-risk leukemia.

Hong-xia ZHAO ; Wan-jun SUN ; Jie LI ; Jun-xiao QIAO ; Ya-jing HUANG ; Hai-lan HU ; Hui-sheng AI

Chinese Journal of Hematology.2013;34(11):922-925. doi:10.3760/cma.j.issn.0253-2727.2013.11.003

OBJECTIVETo evaluate the anti-leukemia effects of prophylactic G-CSF mobilized donor lymphocytes infusion (pG-DLI) and its relationship with the incidence of graft-versus-host disease (GVHD) in high-risk leukemia patients with non-myeloablative stem cell transplantation (NST).

METHODS12 patients with high-risk leukemia were analyzed, including Ph⁺ acute lymphocytic leukemia (n=1), acute leukemia (AL) with persistent non-complete remission (n=2), acute myeloid leukemia (AML) with central nervous system (CNS) relapse (n=3), hybrid AL (n=1), secondary AML evolving from myelodysplastic syndrome (MDS/AML) (n=2), chronic myeloid leukemia in accelerated phase (CML-AP) (n=1), CML in blastic phase (CML-BP) (n=2). All patients received non-myeloablative conditioning and pG-DLIs were administered 30-40 days post transplantation if no signs of GVHD were present. The percentage of donor cell chimera was analyzed by short tandem repeat-polymerase chain reaction (STR-PCR) just before and after pG-DLI. The incidence of leukemia relapse and GVHD were observed.

RESULTS12 high-risk leukemia patients with a median age of 38 (range: 29-52) years received G-DLI at a median interval of 35 (32-40) days. The median numbers of infused mononuclear cells (MNCs), CD34⁺, and CD3⁺ cells/kg recipient body weight was 2.3×10⁸/kg, 1.7×10⁶/kg, and 0.6×10⁸/kg, respectively. 10 of 12 patients had full donor chimera before pG-DLIs and conversion from mixed to full donor chimera occurred in the other 2 patients shortly after pG-DLI. Grade II acute GVHD (aGVHD) was observed in only 2 patients and chronic GVHD (cGVHD) developed in 6 patients, including involvement of skin (n=3), skin and intestine (n=2), liver (n=1). 1 patient died of cGVHD. With a median follow-up of 40 (24-64) months, 7 patients are alive in remission, with 3-year actuarial overall survival (OS) and disease-free survival (DFS) rates of the same 58.3%.

CONCLUSIONOur findings indicate that pG-DLI after NST does not increase the risk of aGVHD, but could enhance the capacity graft-vs-leukemia and prevent relapse in high-risk leukemia patients.

Adult ; Female ; Graft vs Host Disease ; prevention & control ; Granulocyte Colony-Stimulating Factor ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Leukemia ; surgery ; Lymphocyte Transfusion ; methods ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; prevention & control ; Tissue Donors ; Transplantation, Homologous

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Detection of intron 22 inversion of factor VIII gene in severe hemophilia A patients.

Zhi-ping GUO ; Jian-fang CHEN ; Xiu-yu QIN ; Yao-fang ZHANG ; Lin-hua YANG

Chinese Journal of Hematology.2013;34(11):918-921. doi:10.3760/cma.j.issn.0253-2727.2013.11.002

OBJECTIVETo investigate the incidence of intron 22 inversion (INV22) of factor VIII (FVIII) gene in severe hemophilia A (HA) patients, clarify its pathological mechanism, and identify INV22 carrier in the female family members.

METHODSOne-stage method was used to assay the FVIII activity (FVIII:C)in 126 severe HA patients with a median age of 14 years old (range: 4 months-63 years). INV22 was analyzed by long-distance polymerase chain reaction (LD-PCR) and pulsed field gel electrophoresis (PFGE), and pedigree were conducted in 3 involved HA families.

RESULTSOf all the 126 severe HA, 52 (41.3%) cases had the INV22. Four females including 3 mothers and 1 sister of probands were diagnosed as INV22 carriers among 11 suspected carrier mosaicisms from 3 INV22 positive HA families. In 8 females from one family without HA history, the patient's mother was a INV22 carrier, but her maternal grandmother, 2 maternal aunts, 2 female siblings and 1 elder female cousin were negative.

CONCLUSIONLD-PCR and PFGE could be used to diagnose severe HA patients with INV22 and identify the carriers.

Adolescent ; Adult ; Child ; Child, Preschool ; Chromosome Inversion ; Chromosomes, Human, X ; Factor VIII ; genetics ; Female ; Hemophilia A ; genetics ; Heterozygote ; Humans ; Infant ; Introns ; Male ; Middle Aged ; Pedigree

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Clinical characteristics of 520 children with hemophilia in a single medical center.

Min XUAN ; Rong-feng FU ; Feng XUE ; Yan-hui YANG ; Lei ZHANG ; Ren-chi YANG

Chinese Journal of Hematology.2013;34(11):913-917. doi:10.3760/cma.j.issn.0253-2727.2013.11.001

OBJECTIVETo analyze the clinical characteristics, diagnosis and treatment of pediatric hemophilia in single center over the decade.

METHODSA retrospective study was conducted with 520 hemophilic children hospitalized in our medical center between January 2002 and December 2012.

RESULTSAll the patients were male including 438 hemophilia A (HA) and 82 hemophilia B (HB). There were significant differences in APTT between severe and mild- to moderate hemophilia (P<0.05). In pediatric HA and HB, delay time of diagnosis were 1.42 and 1.17 year, respectively. Children of 7-12 years were the largest population of visiting a doctor, and the spontaneous bleeding episode was the main cause. The most common hemorrhage site was soft tissue in early childhood, but joint was increasingly affected with age as children growth. All bleeding sites and frequencies were not associated with plasma factor level of patient (P>0.05). Knee and anKle were mainly involved in early child, while elbow and shoulder were involved increasingly in later childhood. Additionally, in HA and HB, inhibitor occurrence were 8.9%(19/214) and 12.8%(5/39), inducing 78.9%(15/19) and 40.0%(2/5) of high titer inhalator, and antiHCV-positive rate were 2.8%(11/397) and 2.5%(2/79), respectively.

CONCLUSIONOur data highlights that delay in diagnosis and blood-borne infections were significantly reduced over the decade, but the development of inhibitor still remains a major challenge with wide-scale usage of factor in replacement therapy.

Adolescent ; Child, Preschool ; Hemophilia A ; Hemophilia B ; Humans ; Infant ; Male ; Retrospective Studies

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Microtransplantation for treatment of Burkitt lymphoma.

Jie LI ; Wan-jun SUN ; Hong-xia ZHAO ; Na-na DAI ; Hui-sheng AI

Chinese Journal of Hematology.2013;34(10):912-912. doi:10.3760/cma.j.issn.0253-2727.2013.10.021

Burkitt Lymphoma ; therapy ; Hematopoietic Stem Cell Transplantation ; Humans

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Thoughts on differential diagnosis between aplastic anemia and hypoplastic myelodysplastic syndrome.

Jun SHI ; Yi-zhou ZHENG

Chinese Journal of Hematology.2013;34(10):910-912. doi:10.3760/cma.j.issn.0253-2727.2013.10.020

Anemia, Aplastic ; diagnosis ; Diagnosis, Differential ; Humans ; Myelodysplastic Syndromes ; diagnosis

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Expression and significance of MUM1/IRF4 in hematological malignancies.

Sen-sen ZHANG ; Li-juan CHEN

Chinese Journal of Hematology.2013;34(10):907-909. doi:10.3760/cma.j.issn.0253-2727.2013.10.019

Hematologic Neoplasms ; metabolism ; Humans ; Interferon Regulatory Factors ; metabolism

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Diagnosis value of γ-IFN on the specific pathogen infection associated hemophagocytic lymphohistocytosis.

Wei-qun XU ; Bin-hua PAN ; Zen WANG ; Ling-yan ZHANG ; Bei YE ; Yong-min TANG

Chinese Journal of Hematology.2013;34(10):904-906. doi:10.3760/cma.j.issn.0253-2727.2013.10.018

Humans ; Interferon-gamma ; analysis ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; microbiology

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