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

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*

OBJECTIVE: To explore the expression characteristics of antigens and functional markers of natural killer (NK) cells in patients with acute myeloid leukemia (AML). METHODS: Multi-parameter flow cytometry was used to detect NK cell surface markers and their functional indicators in 56 newly diagnosed AML patients and 24 healthy controls, including activating receptors NKG2D, NKP46, DNAM-1, and killing indicators granzyme B, perforin. RESULTS: Referring to the WHO hematopoiesis and lymph tissue tumor classification criteria, 56 cases were roughly divided into three types: AML M₁, M₂, and M₄/M₅. However, there was no differences about NK cells among the three types, so it was no longer subdivided. NK cells were divided into two groups: CD3^-CD56^hiCD16^- (CD56^hiNK) and CD3^-CD56^dimCD16⁺ (CD56^dimNK). Compared with CD56^dimNK cell population, except for NKP46, the positive expression levels of NKG2D and other receptors of CD56^hiNK cells in AML patients decreased (P<0.001). Compared with healthy controls, the proportion of CD56^hiNK cells in AML patients increased, while the number and proportion of NK cells and proportion of CD56^dimNK cells significantly decreased (P<0.05). The proportion of perforin in CD56^hiNK cells significantly increased (P<0.05). The expression of DNAM-1 in CD56^hiNK cells, NKG2D, DNAM-1, and perforin in CD56^dimNK cells decreased significantly (P<0.05). There was no statistically significant difference in expression of other functional indexes in AML patients compared with corresponding indexes of healthy controls. In addition, the proportion of CD56^hiNK cells was positively correlated with the expression of CD34⁺ in AML (r=0.303). CONCLUSION Compared with CD56^dimNK, the ratio of CD56^hiNK and the expression of functional markers in AML patients are lower. Compared with healthy controls, the number and expression ratio of NK cells in AML patients decrease and the expression of functional markers is abnormal, indicating that its function is impaired.
CD56 Antigen ; Flow Cytometry ; Humans ; Killer Cells, Natural ; Leukemia, Myeloid, Acute

OBJECTIVE: To investigate the expression of CD56 in multiple myeloma (MM) cells and its relationship between extramedullary disease and extramedullary relapse. METHODS: Clinical data of 99 patients with MM treated in our hospital from January 2015 to December 2019 was retrospectively analyzed. The patients were divided into positive group and negative group according to the expression of CD56. The relationship between CD56 and multiple myeloma extramedullary disease, extramedullary relapse was analyzed. RESULTS: Among 99 newly diagnosed patients with MM, the positive rate of CD56 was 65%, and the incidence of extramedullary disease of patients in the CD56 positive group was lower than that in the CD56 negative group (17.19% vs 48.57%) (P<0.01). Meanwhile, the incidence of extramedullary relapse of patients in the CD56 positive group was lower than that in the CD56 negative group (1.56% vs 34.29%) (P<0.01). CONCLUSION CD56 is highly expressed in MM, and its low expression is associated with the occurrence of extramedullary disease and extramedullary relapse, which suggests that CD56 may be an important indicator for predicting the occurrence of extramedullary disease and extramedullary relapse.
CD56 Antigen ; Humans ; Multiple Myeloma ; Neoplasm Recurrence, Local ; Retrospective Studies

OBJECTIVE: To investigate the significance of CD27 and CD56 in the prognosis of multiple myeloma (MM) patients, and to establish a simple and convenient prognostic risk score. METHODS: One hundred and eleven newly diagnosed MM patients treated by bortezomib in Shengjing hospital from January 1, 2013 to January 1, 2019 were selected, and the relationship between clinical characteristics and survival time of patients was analyzed. RESULTS: The overall survival (OS) of patients in CD27 CONCLUSION Among patients with MM treated by bortezomib, CD27
Bortezomib ; CD56 Antigen ; Hematopoietic Stem Cell Transplantation ; Humans ; Multiple Myeloma ; Prognosis ; Retrospective Studies

OBJECTIVE: To investigate the expression of costimulatory molecule Tim3 and its subsets on NK cells in patients with acute myeloid leukemia. METHODS: 30 patients with acute myeloid leukemia treated in our hospital from August 2016 to August 2018 were randomly selected as the AML group, and 30 healthy persons in our hospital during the same period were randomly selected as the control group. The expression levels of CD56 RESULTS: The expression levels of CD56 CONCLUSION The expression of costimulatory molecule Tim3 in NK cells and its subsets of patients with acute myeloid leukemia is down-regulated.
CD56 Antigen ; Flow Cytometry ; Hepatitis A Virus Cellular Receptor 2 ; Humans ; Killer Cells, Natural ; Leukemia, Myeloid, Acute ; Patients

Bortezomib/therapeutic use* ; CD56 Antigen ; Humans ; Multiple Myeloma ; Prognosis ; Proto-Oncogene Proteins c-kit

Endometrial stromal sarcoma (ESS) is a rare malignant tumour of the endometrium, accounts for less than 1% of all uterine malignancies. Routinely, it is diagnosed morphologically, supported by immunomarkers of CD10 and vimentin. CD56 is used widely in neuroendocrine tumour. In our current practice, CD56 is not used to support the diagnosis of ESS. We present a case of a postmenopausal lady with advanced ESS who had expression of CD56 upon immunohistochemical study
CD56 ; endometrial stromal sarcoma ; immunohistochemistry ; uterine leiomyoma ; vaginal neoplasm

OBJECTIVETo investigate the clinical significance of RUNX1-RUNX1T1 expression level in bone marrow of patients with acute non-M3 myeloid leukemia (AML non-M3), and to understand the biological characteristics of RUNX1-RUNX1T1 positive AML expressing lymphoid antigens CD19, CD56 and its effect on the initially induced remission rate and prognosis.

METHODSThe expression level of RUNX1-RUNX1T1 in bone marrow of 200 patients with newly diagnosed AML (non-M3) was detected by real-time fluorescent Q-PCR, the expression level of lymphoid antigens was detected by flow cytometry, and the relationship of the initially induced remission rate (CR1) with the overall survival (OS) rate was analyzed, the CR1 and OS differences also were analyzed between CD56 and CD56 patients as well as CD19 and CD17 patients in RUNX1-RUNX1T1 positive patients with AML.

RESULTSThe CD56 patients at the initial diagnosis had lower CR1(P<0.05) in RUNX1-RUNX1T1 positive AML patients, the CR1 of CD19 patients was higher than that in CD19 patients at the initial diagnosis (P<0.05). The OS of CD56 patients was significantly high in comparison with CD56 patients (P<0.05), while the OS between CD19 patients and CD19 patients was not significantly different.

CONCLUSIONThe bone marrow CD56 in RUNX1-RUNX1T1 positive AML patients suggests poor prognosis. The CD19 only correlates with CR1, but does not with OS.

OBJECTIVETo establish a novel method for ex vivo expansion of natural killer cells from human umbilical blood, so as to provide the basis for NK cell therapy.

METHODSMononucleated cells from human umbilical blood were harvested and suspended in a serum-free medium containing 5% autologous plasma, recombinant human IL-15 (50 ng/ml) and hydrocortisone sodium succinate (5×10 mol/L) at a concentration of 1.5×10/ml, then the cells were seeded into flasks pre-coated with heparin sodium (100 U/cm) or/and anti-human CD16 antibody (1 µg/cm). After culture for 2 weeks, the cells were harvested and counted. Ratios of CD3/CD56 of the cells were determined by flow cytometry. MTT test was performed to assess the cytotoxicity against K562 cells with graded ratios of effector/target cells.

RESULTSIn contrast to the cells in flasks without pre-coating, the attached colonies appeared predominantly within 1 week of culture from heparin- and antibody-coated groups. The cell numbers from the pre-coated groups were significantly higher than that of uncoated one after culture for 2 weeks. Furthermore, the ratios of CD3/CD56 cells were much higher in pre-coated groups, and that of the cells from flasks pre-coated with heparin and antibody were the highest (all the P values <0.01). MTT test showed that the cytotoxic activity of the cells stimulated by precoating were much more potent than that of the cells without the stimulation.

CONCLUSIONAdvantageous expansion of NK cells can be achieved by precoating with heparin and anti-CD16 antibody, and also by supplement with IL-15 and hydrocortisone into the media, so the umbilical NK cells with high purity and potent cytotoxicity can be obtained.