OBJECTIVE: To explore the similarities and variations of biological phenotype and cytotoxicity of human umbilical cord blood natural killer cells (hUC- NK) after human umbilical cord blood-derived mononuclear cells (hUC-MNC) activated and expanded by two in vitro high-efficient strategies. METHODS: Umbilical cord blood mononuclear cells (MNC) from healthy donor were enriched by Ficoll-based density gradient centrifugation. Then, the phenotype, subpopulations, cell viability and cytotoxicity of NK cells derived from Miltenyi medium (denoted as M-NK) and X-VIVO 15 (denoted as X-NK) were compared using a "3IL" strategy. RESULTS: After a 14-day's culture, the contents of CD3^-CD56⁺ NK cells were elevated from 4.25%±0.04% (d 0) to 71%±0.18% (M-NK) and 75.2%±1.1% (X-NK) respectively. Compared with X-NK group, the proportion of CD3⁺CD4⁺ T cells and CD3⁺CD56⁺ NKT cells in M-NK group decreased significantly. The percentages of CD16⁺, NKG2D⁺, NKp44⁺, CD25⁺ NK cells in X-NK group was higher than those in the M-NK group, while the total number of expanded NK cells in X-NK group was half of that in M-NK group. There were no significant differences between X-NK and M-NK groups in cell proliferation and cell cycle, except for the lower percentage of Annexin V+ apoptotic cells in M-NK group. Compared with X-NK group, the proportion of CD107a⁺ NK cells in M-NK group were higher under the same effector-target ratio (E∶T) (P<0.05). CONCLUSION The two strategies were adequate for high-efficient generation of NK cells with high level of activation in vitro, however, there are differences in biological phenotypes and tumor cytotoxicity.
Humans ; Fetal Blood ; Killer Cells, Natural ; T-Lymphocytes ; Leukocytes, Mononuclear/metabolism* ; Cell Proliferation ; CD56 Antigen/metabolism*

OBJECTIVETo explore the immunophenotype characteristics of newly diagnosed patients with CD56⁺ acute myeloid leukemia (AML).

METHODSCombining with cytomorphology, four-color flow cytometry was used to analyze the immunophenotype of 342 AML patients with CD56⁺ or CD56⁻.

RESULTSIn 342 AML patients, the CD56⁺ expression was found in 83 AML patients who accounted for 24.27% and included 10 cases of M1, 45 cases of M2, 5 cases of M3, 6 cases of M6 and 17 cases of M5. The statistical analysis showed that there was statistical difference between CD56⁺ and CD11b⁺ patients (P < 0.05), but there was no statistical difference between CD56⁺ and HLA-DR, CD34, CD38, CD13, CD33, CD15, CD117, CD14, CD64, CD2, CD7, CD5, CD3, CD4, CD10, CD19, CD20, CD22 (P > 0.05).

CONCLUSIONAML with only CD56 positive always has poor prognosis, thus the prognosis of patients with CD56⁺ AML accompanied by other antigens still needs more research.

CD56 Antigen ; metabolism ; Flow Cytometry ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; classification ; Prognosis

CD4 Antigens ; metabolism ; CD56 Antigen ; metabolism ; Child ; Humans ; Lymphoma, T-Cell, Cutaneous ; diagnosis ; metabolism ; Male ; Skin Neoplasms ; diagnosis ; metabolism

Adult ; CD56 Antigen ; metabolism ; Carcinoma, Neuroendocrine ; metabolism ; pathology ; surgery ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Mediastinal Neoplasms ; metabolism ; pathology ; surgery ; Mediastinum ; pathology ; surgery ; Phosphopyruvate Hydratase ; metabolism ; Synaptophysin ; metabolism

OBJECTIVE: To investigate the regulatory effect of interferon-α (IFN-α) on the apoptosis and killing function of CD56^dimCD57⁺ natural killer (NK) cells in systemic lupus erythematosus (SLE) patients, and to explore the specific mechanism. METHODS: A total of sixty-four newly treated SLE patients and sixteen healthy controls (HC) enrolled in the Second Hospital of Dalian Medical University were selected as the research subjects. And the gene expression levels of molecules related to NK cell-killing function were detected by real-time quantitative polymerase chain reaction. CD56^dimCD57⁺ NK cells were co-cultured with the K562 cells, and the apoptotic K562 cells were labeled with Annexin-Ⅴ and 7-amino-actinomycin D. Peripheral blood mononuclear cells were treated with 20, 40, and 80 μmol/L hydrogen peroxide (H₂O₂), and treated without H₂O₂ as control, the expression level of perforin (PRF) was detected by flow cytometry. The concentration of IFN-α in serum was determined by enzyme linked immunosorbent assay. The expression levels of IFN-α receptors (IFNAR) on the surface of CD56^dimCD57⁺ NK cells were detected by flow cytometry, and were represented by mean fluorescence intensity (MFI). CD56^dimCD57⁺ NK cells were treated with 1 000 U/mL IFN-α for 24, 48 and 72 h, and no IFN-α treatment was used as the control, the apoptosis and the expression levels of mitochondrial reactive oxygen species (mtROS) were measured by flow cytometry and represented by MFI. RESULTS: Compared with HC(n=3), the expression levels of PRF1 gene in peripheral blood NK cells of the SLE patients (n=3) were decreased (1.24±0.41 vs. 0.57±0.12, P=0.05). Compared with HC(n=5), the ability of peripheral blood CD56^dimCD57⁺ NK cells in the SLE patients (n=5) to kill K562 cells was significantly decreased (58.61%±10.60% vs. 36.74%±6.27%, P < 0.01). Compared with the control (n=5, 97.51%±1.67%), different concentrations of H₂O₂ treatment significantly down-regulated the PRF expression levels of CD56^dimCD57⁺ NK cells in a dose-dependent manner, the 20 μmol/L H₂O₂ PRF was 83.23%±8.48% (n=5, P < 0.05), the 40 μmol/L H₂O₂ PRF was 79.53%±8.56% (n=5, P < 0.01), the 80 μmol/L H₂O₂ PRF was 76.67%±7.16% (n=5, P < 0.01). Compared to HC (n=16), the serum IFN-α levels were significantly increased in the SLE patients (n=45) with moderate to high systemic lupus erythematosus disease activity index (SLEDAI≥10) [(55.07±50.36) ng/L vs. (328.2±276.3) ng/L, P < 0.001]. Meanwhile, compared with HC (n=6), IFNAR1 expression in peripheral blood CD56^dimCD57⁺ NK cells of the SLE patients (n=6) were increased (MFI: 292.7±91.9 vs. 483.2±160.3, P < 0.05), and compared with HC (n=6), IFNAR2 expression in peripheral blood CD56^dimCD57⁺ NK cells of the SLE patients (n=7) were increased (MFI: 643.5±113.7 vs. 919.0±246.9, P < 0.05). Compared with control (n=6), the stimulation of IFN-α (n=6) significantly promoted the apoptosis of CD56^dimCD57⁺ NK cells (20.48%±7.01% vs. 37.82%±5.84%, P < 0.05). In addition, compared with the control (n=4, MFI: 1 049±174.5), stimulation of CD56^dimCD57⁺ NK cells with IFN-α at different times significantly promoted the production of mtROS in a time-dependent manner, 48 h MFI was 3 437±1 472 (n=4, P < 0.05), 72 h MFI was 6 495±1 089 (n=4, P < 0.000 1), but there was no significant difference at 24 h of stimulation. CONCLUSION High serum IFN-α level in SLE patients may induce apoptosis by promoting mtROS production and inhibit perforin expression, which can down-regulate CD56^dimCD57⁺ NK killing function.
Humans ; Interferon-alpha/metabolism* ; Perforin/metabolism* ; Leukocytes, Mononuclear/metabolism* ; Hydrogen Peroxide/metabolism* ; Interferon-gamma/metabolism* ; CD56 Antigen/metabolism* ; Killer Cells, Natural/metabolism* ; Lupus Erythematosus, Systemic

This study was aimed to investigate the clinical features of CD56(+) patients with acute monocytic leukemia (AML-M5) and their prognostic significance. The data of 76 newly-diagnosed patients from our hospital were analyzed retrospectively. Patients were divided into two groups: CD56(+) group (21 patients) and CD56(-) group (55 patients). The clinical features, CR rate, relapse rate, the duration of CR, and survival time of patients between the two groups were compared. The results indicated that the CD56(+) antigen was observed in 21 patients (27.6%), their median age was 51.5 years and with a range 16 - 70 years. Of the 21 CD56(+) patients, the high WBC count was found in 57.1% CD56(+) patients (12/21), but it only in 15% CD56(-) patients (P < 0.05). The extramedullary infiltration was seen in 13 CD56(+) patients, and accounted for 62% (13/21), meanwhile this infiltration was found in 18 CD56(-) patients (18/55) and accounted for 33% (P < 0.05). All cases immunophenotypically highly expressed CD13, CD33, CD64, CD11b, cMPO, CD38, in which only the expression frequency of CD11b was positively related with CD56 (r = 0.59, P < 0.05). The CR rate in CD56(+) group accounted for 60.0%, and had no significant difference in comparison with that in CD56(-) group. In CD56(+) group the relapse rate was 75% (P = 0.042), the mean duration of CR was 5.5 months (95%CI, 3.1 - 8.6, P = 0.002), the median overall survival time was 10.1 months (95%CI, 2.3 - 16.3, P = 0.001). and all these had statistical significance as compared with that in CD56(-) group. It is concluded that CD56(+) AML-M5 patients always complicate with high WBC count and extramedullary infiltration, their CR rate and duration of CR are lower and shorter respectively, their relapse rate and prognosis are high and poor respectively.
Adolescent ; Adult ; Aged ; CD56 Antigen ; metabolism ; Female ; Humans ; Immunophenotyping ; Leukemia, Monocytic, Acute ; diagnosis ; immunology ; Male ; Middle Aged ; Prognosis ; Young Adult

OBJECTIVETo characterize the phenotypic and biological properties of CD56(+) natural killer cells from human peripheral blood mononuclear cells (PBMCs).

METHODSSurface markers and intracellular cytotoxic molecules were stained with multi-color-labeled monoclonal antibodies and analyzed at the single cell level the relation between NK subsets and biological characteristics by flow cytometry.

RESULTSNK cells in PBMCs could be divided into two major populations, CD56(bright) and CD56(dim), based upon the expression of CD56 molecules. Both CD56(bright) and CD56(dim) expressed CD95 (Fas) with CD95(bright) and CD95(dim) subsets. CD56(dim) subsets had higher percentage of CD8, granzyme B and perforin expression compared to those of CD56(bright) subsets. In CD56(bright) and CD56(dim) subpopulations, CD95(bright) and CD8(+) subsets had higher percentage of granzyme B and perforin expression.

CONCLUSIONCD56(+) NK cells in PBMCs are composed of distinct subpopulations, CD56(dim) and CD56(dim) CD8(+) NK subsets have higher percentage of granzyme B and perforin and may play an important role in the killing of target cells.

CD56 Antigen ; metabolism ; CD8 Antigens ; metabolism ; Granzymes ; metabolism ; Humans ; Killer Cells, Natural ; classification ; immunology ; metabolism ; Lymphocyte Subsets ; immunology ; Perforin ; metabolism ; Phenotype ; fas Receptor ; metabolism

OBJECTIVE: To identify the function of neural cell adhesion molecule (NCAM) mutation in the genesis of human glioma. METHODS: Mutations were found through polymerase chain reaction-single strand conformation polymorphism, and then the changed DNA fragments were purified and multiplied and sent to Shanghai for sequencing. Blast and ORF finder were used to find out the amino changes in NCAM. RESULTS: An A-C transversion was found at position 1 126 in NCAM's 7 exon in a patient with glioblastoma from 43 astrocytoma. CONCLUSION Structural change in the protein caused by point mutation may be the reason for tumorigenesis of astrocytoma.
Adult ; Astrocytoma ; genetics ; metabolism ; Base Sequence ; Brain Neoplasms ; genetics ; metabolism ; CD56 Antigen ; genetics ; DNA Mutational Analysis ; Female ; Humans ; Male ; Molecular Sequence Data ; Point Mutation

OBJECTIVETo investigate the expressions of cell surface differentiation antigen CD56 and CD11b antigen in acute monocytic leukemic (AML-M(5)) cells and their clinical significance.

METHODSA total of 113 cases of de nove adult AML-M(5) were examined genetically and immunologically using G-banding technique, interphase fluorescence in situ hybridization (I-FISH) and flow cytometry immunophenotyping, and the results were analyzed in relation to their clinical data.

RESULTSOf the 113 cases, the expression rates of CD56 and CD11b was 28.32% and 73.45%, respectively. The CD56(+) patients had high CD11b expression, and the expression levels of CD11b and CD56 were positively correlated (P<0.05). The incidence of karyotypic abnormalities was 48.57% (55 cases) in these patients, including 25 (22.12%) with 11q23 aberrations. Twenty-five cases were positive for MLL gene abnormalities as found by I-FISH analysis. Compared with the patients positive for both CD56 and CD11b, those negative for both CD56 and CD11b showed increased peripheral blood white blood cell (WBC) count and also increased blast and progenitor cells in the bone marrow (P<0.05); the former patients often had karyotypic abnormalities, commonly involving 11q23 aberrations (P<0.05), whereas the latter patients presented more likely with extramedullary infiltration and refractory leukemia (P<0.01) with lowered complete remission rate and shortened median survival time (P<0.01). CD56-positive patients were more likely to have karyotypic abnormalities and refractory leukemia than CD11b-postive patients (P<0.05), but the peripheral blood WBC counts, bone marrow blast and progenitor cells, extramedullary infiltration, complete remission rate or median survival time showed no significant differences between them (P>0.05).

CONCLUSIONAML-M(5) patients with CD56 positivity and high expression of CD11b often have aberrant karyotypes, commonly involving 11q23/MLL gene abnormality. These patients frequently develop extramedullary infiltration and refractory leukemia often with poor prognosis.

Adolescent ; Adult ; Aged ; CD11b Antigen ; genetics ; metabolism ; CD56 Antigen ; genetics ; metabolism ; Female ; Gene Expression Regulation ; Humans ; Karyotyping ; Leukemia, Monocytic, Acute ; diagnosis ; genetics ; metabolism ; pathology ; Leukocyte Count ; Male ; Middle Aged ; Prognosis ; Young Adult

Adult ; CD56 Antigen ; metabolism ; Carcinoma ; metabolism ; pathology ; surgery ; Carcinoma, Medullary ; pathology ; Carcinoma, Papillary ; metabolism ; pathology ; DNA-Binding Proteins ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Paraganglioma ; metabolism ; pathology ; Thyroid Neoplasms ; metabolism ; pathology ; surgery ; Thyroidectomy ; methods ; Transcription Factors ; Triglycerides ; metabolism