Regulatory effect of interleukin-18 on natural killer cell activity in patients with alopecia areata / 中华皮肤科杂志

Objective:To investigate changes of natural killer (NK) cell subsets and interleukin-18 (IL-18) level in peripheral blood of patients with alopecia areata, and to assess the regulatory effect of IL-18 on NK cell activity.Methods:A total of 67 patients with alopecia areata (alopecia areata group) and 25 healthy volunteers (control group) were collected from Shanxi Provincial People′s Hospital between December 2019 and January 2021. Peripheral blood mononuclear cells (PBMCs) and plasma were isolated. The percentage of NK cell subsets was investigated by flow cytometry, and plasma IL-18 level was measured by enzyme-linked immunosorbent assay. PBMCs were stimulated with recombinant human IL-18, and co-culture systems of PBMCs with 721.221 cells, K562 cells and P815-Ab cells were established separately. NK cell function was assessed by determining the percentage of CD107a-expressing NK cells and fluorescence intensity of CD16 + NK cells. Comparisons between groups were performed using t test or paired t test. Results:Compared with the control group, the alopecia areata group showed significantly decreased percentage of CD56 +CD16 - NK cells (8.12% ± 3.14% vs. 10.78% ± 4.08%, t = 3.33, P = 0.001) , but significantly increased percentage of CD56 +CD16 + NK cells (46.08% ± 15.21% vs. 32.14% ± 10.45%, t = 4.22, P < 0.001) , and there was no significant difference in the percentage of CD56 -CD16 + NK cells between the alopecia areata group and control group (28.81% ± 8.65% vs. 27.09% ± 7.62%, t = 0.88, P = 0.383) . The plasma IL-18 level was significantly higher in the alopecia areata group than in the control group (112.0 ± 23.72 pg/ml vs. 99.34 ± 15.15 pg/ml, t = 2.48, P = 0.015) . After co-culture with 721.221 cells, K562 cells and P815-Ab cells, the percentage of CD107a-expressing NK cells was significantly higher in NK cells from the alopecia areata group (9.53% ± 1.70%, 5.15% ± 1.35%, 6.50% ± 1.64%, respectively) than in those from the control group (5.00% ± 1.17%, 4.40% ± 1.09%, 5.13% ± 1.36%, respectively, all P < 0.05) . After the stimulation with P815-Ab cells, the alopecia areata group showed significantly decreased fluorescence intensity of CD16 + NK cells (151.10% ± 59.30%) compared with the control group (221.90% ± 93.56%, t = 4.31, P < 0.001) . After IL-18 stimulation, the percentage of CD107a-expressing NK cells significantly increased in the co-culture system of NK cells with 721.221 cells compared with the unstimulated co-culture system (14.47% ± 2.67% vs. 9.93% ± 1.94%, t = 6.00, P < 0.001) , while there was no significant difference between the IL-8-stimulated co-culture system of NK cells with K562 cells or P815-Ab cells and the unstimulated co-culture systems (both P > 0.05) . Conclusion:IL-18 could enhance NK cell activity in patients with alopecia areata, likely by promoting natural cytotoxicity receptor-mediated cytotoxicity.

Effect of naringenin on natural killer cell cytotoxicity against hepatocellular carcinoma cells in vitro and its mechanism / 临床肝胆病杂志

Objective To investigate the effect of naringenin on the killing rate of natural killer (NK) cells and related mechanism by amplification of human peripheral blood mononuclear cells into NK cells in vitro and co-culture with hepatocellular carcinoma (HCC) CLC5 cells at a ratio of 1∶ 1. Methods A lymphocyte separation medium was used to isolate human peripheral blood mononuclear cells, which were induced with recombinant human interleukin-2 in vitro to culture NK cells. CCK-8 assay was used to measure the proliferation of HCC cells after human HCC cells were treated with naringenin (0, 3.125, 6.25, 12.5, 25, and 50 μmol/L) for 0, 24, and 48 hours, and after human NK cells were treated with different concentrations of naringenin for 24 hours, CCK-8 assay was used to measure the proliferation of NK cells. CellTiter-LumiTM was used to measure the killing rate of NK cells after the NK-HCC cell co-culture system at the ratio of 1∶ 1 was treated with naringenin for 24 hours. Quantitative real-time PCR was used to measure the gene expression of the activating receptor NKG2D in NK cells and NKG2D ligands in HCC cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results After being induced and cultured by recombinant human interleukin-2, NK cells were amplified to 82.33%±0.70% of human peripheral blood mononuclear cells. After naringenin treatment for 24 hours, there was no significant difference in the proliferation rate of HCC CLC5 cells between all mass concentration groups (all P > 0.05), and in the 25 and 50 μmol/L mass concentration groups, naringenin significantly promoted the proliferation of NK cells (both P 0.05); it significantly upregulated the expression of the NKG2D ligands such as ULBP1 and ULBP3 in HCC cells (all P < 0.001). Conclusion Naringenin may increase the killing activity of NK cells by upregulating the expression of NKG2D ligands in HCC cells.

Invariant Natural Killer T-cells and their subtypes may play a role in the pathogenesis of endometriosis

Abstract Objective To evaluate the frequencies of iNKT cells and their subsets in patients with deep endometriosis. Methods A case-control study was conducted between 2013 and 2015, with 73 patients distributed into two groups: 47 women with a histological diagnosis of endometriosis and 26 controls. Peripheral blood, endometriosis lesions, and healthy peritoneal samples were collected on the day of surgery to determine the frequencies of iNKT cells and subtypes via flow cytometry analysis. Results The authors observed a lower number of iNKT (p= 0.01) and Double-Negative (DN) iNKT cells (p= 0.02) in the blood of patients with endometriosis than in the control group. The number of DN iNKT IL-17+ cells in the secretory phase was lower in the endometriosis group (p= 0.049). There was an increase in the secretion of IL-17 by CD4+ iNKT cells in the blood of patients with endometriosis and severe dysmenorrhea (p= 0.038), and severe acyclic pelvic pain (p= 0.048). Patients with severe dysmenorrhea also had a decreased number of CD4+ CCR7+ cells (p= 0.022). Conclusion The decreased number of total iNKT and DN iNKT cells in patients with endometriosis suggests that iNKT cells play a role in the pathogenesis of endometriosis and can be used to develop new diagnostic and therapeutic agents.

Role of CXCR6-mediated activation of natural killer T cells in renal fibrosis following acute kidney injury in mice / 中华麻醉学杂志

Objective:To evaluate the role of CXC chemokine receptor 6 (CXCR6)-mediated activation of natural killer T (NKT) cells in renal fibrosis following acute kidney injury (AKI) in mice.Methods:Eighteen male wild-type C57BL/6 mice and 18 CXCR6 knockout C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 6 groups ( n=6 each) using a random number table method: wild-type mouse control group (group WT-CON), CXCR6 knockout mouse control group (group CXCR6 -/--CON), wild-type mouse with AKI group (group WT-AKI), CXCR6 knockout mouse with AKI group (group CXCR6 -/--AKI), wild-type mouse with AKI + NKT cell adoptive transfer group (group WT-AKI-NKT) and CXCR6 knockout mouse with AKI + NKT cell adoptive transfer group (group CXCR6 -/--AKI-NKT). Folic acid 250 mg/kg was intraperitoneally injected to establish the model of renal fibrosis in mice with AKI.NKT cellsuspension 250 μl(1×10 6 cells) was injected through the tail vein on the 4th and 9th days after folic acid injection in group WT-AKI-NKT and group CXCR6 -/--AKI-NKT, respectively.Blood samples were taken from orbital at day 14 after folic acid injection for determination of the concentrations of serum blood urea nitrogen (BUN) and creatinine (Cr). The animals were sacrificed, and renal tissues were obtained for observation of the area of renal fibrosis (by Sirius red staining) and renal injury (using H&E staining) which was scored and for determination of the proportion of CD1d Tetramer+ cells (by flow cytometry), the number of CD206 and α-smooth muscle actin (α-SMA) double positive (CD206 + -α-SMA + ) cells (by immunofluorescence) and expression of interleukin (IL)-4 and IL-13 mRNA (by real-time polymerase chain reaction). Results:Compared with group WT-CON, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI and WT-AKI-NKT ( P<0.05). Compared with group WT-AKI, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI-NKT ( P<0.05), and the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly decreased, and the expression of IL-4 and IL-13 mRNA was down-regulated in group CXCR6 -/--AKI ( P<0.05). Compared with group CXCR6 -/--CON, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased in group CXCR6 -/--AKI and group CXCR6 -/--AKI-NKT ( P<0.05). Compared with group CXCR6 -/--AKI, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group CXCR6 -/--AKI-NKT ( P<0.05). Conclusion:CXCR6-mediated activation of NKT cells is involved in renal fibrosis following AKI in mice, and the mechanism may be related to promoting Th2 cytokine-mediated M2 macrophage-myofibroblast transformation.

The role and mechanism of invariant natural killer T cells in liver anti-tumor immunity / 临床肝胆病杂志

Invariant natural killer T (iNKT) cells are a T lymphocyte subset derived from the thymus and can express both natural killer cell-related receptors and T cell receptor. The iNKT cells are widely distributed in the body and are enriched in the liver, and they exhibit unique functional characteristics and can secrete cytokines and regulate the activity of other immune cells in microenvironment, so as to achieve the role of immune surveillance and disease prevention; especially in tumor microenvironment, iNKT cells can stimulate anti-tumor immune response and reverse immunosuppressive microenvironment in the liver. This article reviews the biological characteristics of iNKT cells and their special role in liver immune homeostasis, especially the anti-tumor effect and mechanism of iNKT cells.

NKT cells in liver diseases / 医学前沿

Natural killer T cells are innate-like and tissue-resident lymphocytes, which recognize lipid antigens and are enriched in the liver. Natural killer T cells play important roles in infections, tumors, autoimmune diseases, and metabolic diseases. In this study, we summarize recent findings on biology of natural killer T cells and their roles in hepatitis B virus and hepatitis C virus infection, autoimmune liver diseases, alcoholic liver disease, nonalcoholic fatty liver disease, and hepatocellular carcinoma. Controversial results from previous studies are discussed, and indicate the dynamic alteration in the role of natural killer T cells during the progression of liver diseases, which might be caused by changes in natural killer T subsets, factors skewing cytokine responses, and intercellular crosstalk between natural killer T cells and CD1d-expressing cells or bystander cells.

Expansion and Sub-Classification of T Cell-Dependent Antibody Responses to Encompass the Role of Innate-Like T Cells in Antibody Responses

In addition to T cell-dependent (TD) Ab responses, T cells can also regulate T cell-independent (TI) B cell responses in the absence of a specific major histocompatibility complex (MHC) class II and antigenic peptide-based interaction between T and B cells. The elucidation of T cells capable of supporting TI Ab responses is important for understanding the cellular mechanism of different types of TI Ab responses. Natural killer T (NKT) cells represent 1 type of helper T cells involved in TI Ab responses and more candidate helper T cells responsible for TI Ab responses may also include γδ T cells and recently reported B-1 helper CD4⁺ T cells. Marginal zone (MZ) B and B-1 cells, 2 major innate-like B cell subsets considered to function independently of T cells, interact with innate-like T cells. Whereas MZ B and NKT cells interact mutually for a rapid response to blood-borne infection, peritoneal memory phenotype CD49d(high)CD4⁺ T cells support natural Ab secretion by B-1 cells. Here the role of innate-like T cells in the so-called TI Ab response is discussed. To accommodate the involvement of T cells in the TI Ab responses, we suggest an expanded classification of TD Ab responses that incorporate cognate and non-cognate B cell help by innate-like T cells.

New method of non-alcoholic fatty liver disease mouse models and mechanism about immune cells / 中国药理学通报

Aim To establish non-alcoholic fatty liver disease mouse model and study different kinds of lymphocytes in C57BL/6J mouse model. Methods SPF male C57BL/6J mice were randomly divided into control group ( normal diet ) and model group( normal diet with high fat diet by gavage) . Models of non-alcoholic fatty liver disease were established. At 12th and 16th weeks, body weight, liver index, serum TC, TG, HDL, LDL, ALT, AST were measured. Pathological examination of fat deposition in liver was performed. Flow cytometry was used to assay the percentage of natural killer cells, T helper cells, natu-ral killer T cells and IL4 +NKT cells in mouse liver. Results Liver index, serum TC, LDL, ALT, AST were significantly higher in model group(P<0.05) after 16 weeks. Pathological sections showed that liver fat deposition in model group was quite severe and large lipid droplets spread through the mouse liver. The percentage of natural killer T cells increased significantly( P<0.05 ) and the percentage of IL4 +NKT cells increased even more obviously(P<0.01). Conclusions C57BL/6J mice fed with normal diet and high fat diet by gavage can form a good non-alcoholic fatty liver disease mouse model. In this model, the number and activity of natural killer T cells are significantly changed, and natural killer T cells may be the new target of the mechanism and drug treatment of nonalcoholic fatty liver dis-ease.

Characteristics of T-and NK/T-cell lymphomas with different pathological subtypes on 18F-FDG PET/CT / 中华核医学与分子影像杂志

Objective To investigate the imaging presentation of T-and NK/T-cell lymphomas with different pathology subtypes on 18F-FDG PET/CT.Methods A total of 95 patients (66 males, 29 females, average age 38.42 years) with T-and NK/T-cell lymphoma proved by pathology from June 2006 to February 2016 were retrospectively analyzed.18F-FDG uptake (SUVmax), nodal invasion, nodal distribution, extra-nodal involvement and staging were compared among 7 pathological subtypes of T-and NK/T-cell lymphomas.One-way analysis of variance, Fisher exact test and Kruskal-Wallis H test were used for data analysis.Results There were significant differences in terms of 18F-FDG uptake, nodal invasion, nodal distribution, extra-nodal involvement and staging among different pathological subtypes of T-and NK/T-cell lymphomas (F=2.937, P<0.05;Fisher exact test,all P<0.01;H=19.883, P<0.01).NK/T-cell lymphoma was found to be prone to invade the nasal cavity and nasopharynx, enteropathic type T-cell lymphoma was specific to the intestine, and subcutaneous panniculitis-like T-cell lymphoma presented with subcutaneous infiltration.All those 3 subtypes were quite specific in their extra-nodal involvement.Most patients with angioimmunoblastic T-cell lymphoma (ATCL), peripheral unspecified T-cell lymphoma (PUTCL) and T immunoblastic lymphoma (TIBL) presented as stage Ⅳ disease.Widespread lymph node disease associated with splenic, parotid and serous membrane involvement were often seen in ATCL patients (most commonly to involve the parotid glands and serous membrane among the 7 subtypes).Nodal involvement was found in PUTCL patients, but extranodal involvement was rather non-specific.TIBL had a non-specific pattern of nodal involvement with low 18F-FDG uptake, lower than ATCL and the other 5 subtypes.Anaplastic large cell lymphoma subtypes had the highest 18F-FDG uptake when compared with the other 6 subtypes, but were less often to manifest as stage Ⅳ disease despite their preponderance for marrow and nodal infiltration.Conclusion Different pathological subtypes of T-and NK/T-cell lymphoma manifest different imaging presentations on 18F-FDG PET/CT, which are useful for understanding their biological characteristics.

Unusual Suspects in the Development of Obesity-Induced Inflammation and Insulin Resistance: NK cells, iNKT cells, and ILCs

The notion that obesity-induced inflammation mediates the development of insulin resistance in animal models and humans has been gaining strong support. It has also been shown that immune cells in local tissues, in particular in visceral adipose tissue, play a major role in the regulation of obesity-induced inflammation. Specifically, obesity increases the numbers and activation of proinflammatory immune cells, including M1 macrophages, neutrophils, Th1 CD4 T cells, and CD8 T cells, while simultaneously suppressing anti-inflammatory cells such as M2 macrophages, CD4 regulatory T cells, regulatory B cells, and eosinophils. Recently, however, new cell types have been shown to participate in the development of obesity-induced inflammation and insulin resistance. Some of these cell types also appear to regulate obesity. These cells are natural killer (NK) cells and innate lymphoid cells (ILCs), which are closely related, and invariant natural killer T (iNKT) cells. It should be noted that, although iNKT cells resemble NK cells in name, they are actually a completely different cell type in terms of their development and functions in immunity and metabolism. In this review, we will focus on the roles that these relatively new players in the metabolism field play in obesity-induced insulin resistance and the regulation of obesity.

Lineage Differentiation Program of Invariant Natural Killer T Cells

Invariant natural killer T (iNKT) cells are innate T cells restricted by CD1d molecules. They are positively selected in the thymic cortex and migrate to the medullary area, in which they differentiate into 3 different lineages. Promyelocytic leukemia zinc finger (PLZF) modulates this process, and PLZFhigh, PLZFintermediate, and PLZFlow iNKT cells are designated as NKT2, NKT17, and NKT1 cells, respectively. Analogous to conventional helper CD4 T cells, each subset expresses distinct combinations of transcription factors and produces different cytokines. In lymphoid organs, iNKT subsets have unique localizations, which determine their cytokine responses upon antigenic challenge. The lineage differentiation programs of iNKT cells are differentially regulated in various mice strains in a cell-intrinsic manner, and BALB/c mice contain a high frequency of NKT2 cells. In the thymic medulla, steady state IL-4 from NKT2 cells directly conditions CD8 T cells to become memory-like cells expressing Eomesodermin, which function as premade memory effectors. The genetic signature of iNKT cells is more similar to that of γδ T cells and innate lymphoid cells (ILCs) than of conventional helper T cells, suggesting that ILCs and innate T cells share common developmental programs.

Deficiencies of Circulating Mucosal-associated Invariant T Cells and Natural Killer T Cells in Patients with Multiple Trauma

Mucosal-associated invariant T (MAIT) cells and natural killer T (NKT) cells are known to play important roles in autoimmunity, infectious diseases and cancers. However, little is known about the roles of these invariant T cells in multiple trauma. The purposes of this study were to examine MAIT and NKT cell levels in patients with multiple trauma and to investigate potential relationships between these cell levels and clinical parameters. The study cohort was composed of 14 patients with multiple trauma and 22 non-injured healthy controls (HCs). Circulating MAIT and NKT cell levels in the peripheral blood were measured by flow cytometry. The severity of injury was categorised according to the scoring systems, such as Acute Physiology and Chronic Health Evaluation (APACHE) II score, Simplified Acute Physiology Score (SAPS) II, and Injury Severity Score (ISS). Circulating MAIT and NKT cell numbers were significantly lower in multiple trauma patients than in HCs. Linear regression analysis showed that circulating MAIT cell numbers were significantly correlated with age, APACHE II, SAPS II, ISS category, hemoglobin, and platelet count. NKT cell numbers in the peripheral blood were found to be significantly correlated with APACHE II, SAPS II, and ISS category. This study shows numerical deficiencies of circulating MAIT cells and NKT cells in multiple trauma. In addition, these invariant T cell deficiencies were found to be associated with disease severity. These findings provide important information for predicting the prognosis of multiple trauma.

Study on the immunoregulatory and lethal effects of natural killer T cells on melanoma cells in vitro / 国际检验医学杂志

Objective To explore the immunoregulatory and lethal effects of natural killer T lymphocytes(NKTs) in vitro .Meth‐ods The mixed lymphocyte cultured(MLC) system was established ,in which the B16F10‐luc‐G5 cells were set as target cells ,the total lymphocyte cells were set as effector cells .(1)In the experiment on immunoregulatory effects ,NKT lymphocytes or CD4+CD25+ T lymphocytes were set as regulating cells ,there was three groups ,including the NKT group ,CD4+CD25+ T group and pure target cell control group .Otherewise ,the 1640 blank control group was set by only adding RPMI1640 solution .(2)In the ex‐periment on antitumor effects ,the NKT or natural killer(NK) lymphocytes were set as killer cells ,there was three groups ,inclu‐ding the NKT group ,NK group and pure target cell control group .Mixed culturing 24 ,48 and 72 hours ,the bioluminescence of target cells in MCL system was detected by using the in vivo imaging system .Results (1)In the experiment on immunoregulatory effects ,there were statistically significant differences in measured average photon numbers between NKT group ,CD4+ CD25+ T group and the two control groups(P<0 .05) .The statistically significant differences were also found in the NKT group between 24 hours and 72 hours (P<0 .05) .(2)In the experiment on antitumor effects ,there were statistically significant differences in meas‐ured average photon numbers ,when the NKT group and NK group were compared to the pure target cell control group(P<0 .05) . After culturing 24 and 72 hours ,statistically significant differences were found between NKT group and NK group(P<0 .05) .Con‐clusion The NKT cells could inhibit the lethal effects of lymphocyte cells on target cells ,and the inhibitory effects are changed by the length of culturing .Compared with the CD4+CD25+ T lymphocytes ,NKT lymphocytes have strongger regulatory effects .Addi‐tionally ,the NKT cells have lethal effects on target cells ,which might be weaker than that of NK cells .

Effects of DC loaded α-GalCer combined with tumor specific cytotoxic T lymphocytes on the growth of transplanted Heps hepatoma in mice / 国际肿瘤学杂志

Objective To investigate the effects of dendritic cells (DCs)loading alpha-Galactosylce-ramide (α-GalCer)combined with tumor specific cytotoxic T lymphocytes (CTLs)on the growth of transplanted Heps hepatoma in mice.Methods We induced the augmentation of the DC cells and T lymphocyte derived from the mice bone marrow,and enabled them to be specific CTLs.DC cells loaded α-GalCer in vitro.First we established a Heps liver cancer xenograft model,then divided the model mice into 4 groups by random number table method (n =9):control group (intravenous injection with physiological saline),CTL group,DC loadingα-Galcer group and DC loading α-Galcer combined with CTLs group.After 2 weeks of intervention,we extrac-ted the tumor tissue,weighed the tumor and calculated the inhibition rate of tumor.The expressions of Bax/Bcl-2 cells in groups of transplanted tumor tissues were detected using immunohistochemistry and Western blotting. Results The average tumor weight of CTL group,DC loading α-GalCer group and combined treatment group were (1 .07 ±0.1 5)g,(1 .1 1 ±0.1 7)g,(0.79 ±0.1 4)g,respectively.All of them were lower than that of control group (1 .69 ±0.23)g,with significant differences (t =1 4.1 76,P =0.023;t =1 2.351 ,P =0.034;t =1 8.672,P =0.000).The average tumor weight of combined treatment group was lower than those of the CTL group and DC loading α-GalCer group,with significant differences (t =1 5.236,P =0.01 2;t =1 1 .1 76, P =0.037).Compared to the CTL group (36.69%)and DC loading α-GalCer group (34.32%),the com-bined treatment group had a higher tumor inhibition rate (53.25%;P =0.034,P =0.021 ).Immunohisto-chemical assay showed that the numbers of Bax-positive cells in CTL group,DC loading α-GalCer group and combined treatment group were 35.83 ±0.75,33.67 ±0.82,41 .1 7 ±1 .1 7 respectively,and compared with the control group (21 .67 ±2.1 6),the differences were statistically significant (t =-1 3.789,P =0.002;t =-1 5.1 1 6,P =0.001 ;t =-1 7.452,P =0.000).The numbers of Bax-positive cells in combined treatment group were different with CTL group and DC loading α-GalCer group (t =-7.730,P =0.009;t =-5.872, P =0.01 1 ).The numbers of Bcl-2-positive cells in CTL group,DC loading α-GalCer group and combined treatment group were 30.83 ±0.75,31 .67 ±1 .03,25.00 ±0.89,and compared with the control group (38.67 ±1 .21 ),the differences were statistically significant (t =9.234,P =0.007;t =1 1 .738,P =0.003;t =20.608,P =0.000).The numbers of Bcl-2-positive cells in combined treatment group were different with CTL group and DC loading α-GalCer group (t =1 1 .952,P =0.003;t =1 2.223,P =0.002).Western blot-ting test results showed that the expression levels of Bax in CTL group,DC loading α-GalCer group and com-bined treatment group were 0.46 ±0.01 ,0.42 ±0.03,0.55 ±0.01 ,and compared with the control group (0.31 ±0.02),the differences were statistically significant (t =1 .035,P =0.032;t =1 .1 24,P =0.027;t =1 .425,P =0.01 0).The expression level of Bax in combined treatment group was different with CTL group and DC loading α-GalCer group (t =1 .305,P =0.01 3;t =1 .421 ,P =0.01 0).The positive expressions of Bcl-2 in CTL group,DC loading α-GalCer group and combined treatment group were 0.34 ±0.03,0.33 ± 0.02,0.24 ±0.01 ,and compared with the control group (0.46 ±0.01 ),the differences were statistically sig-nificant (t =-1 .1 23,P =0.025;t =-1 .061 ,P =0.031 ;t =1 .278,P =0.01 4);the positive expression level of Bcl-2 in combined treatment group was different with CTL group and DC loading α-GalCer group (t =1 .1 60,P =0.021 ;t =1 .21 9,P =0.01 5).Conclusion It has synergistic killing effect on transplanted Heps hepatoma in mice using DC loading α-GalCer combined with the tumor specific CTL.

The clinical significance of T lymphocyte subset in the children patients with community acquired ;pneumonia / 中国医师进修杂志

Objective To observe the effect of T cell subset in the children patients with community acquired pneumonia. Methods A total of 36 patients with community acquired pneumonia were enrolled and they were divided into 2 groups according to pathogen, bacterial infection group, and non-bacterial infection group. And in another way they were divided into severe cases group and non-severe cases group. Indexes of T lymphocyte subset CD3+, CD4+, CD8+, CD19+and activity of natural killer (NK) cell were detected in all patients by flow cytometry and compared. Results The levels of CD3+, CD4 +, CD8 +, CD19 + and NK cell had no significant difference between bacterial infection group and non-bacterial infection group (P>0.05). The levels of CD3+, CD4+and CD19+had no significant difference between severe cases group and non-severe cases group (P>0.05). But the level of CD8+ in severe cases group was significantly higher than that in non-severe cases group:(28.4 ± 7.8)%vs. (14.4 ± 3.5)%, P<0.01. The level of NK cell in severe cases group was significantly lower than that in non-severe cases group: (7.3 ± 2.1)%vs. (16.6 ± 5.4)%, P<0.01. Conclusions The children patients with community acquired pneumonia patients may develop into severe pneumonia with high percent of CD8+or low activity of NK cell. So they should be given immune intervention as soon as possible.

Research on natural killer T cells anergy / 国际外科学杂志

Natural killer T(NKT) cells are a heterogeneous group T cells that share properties of both natural killer cells and T cells,play an important role in tumor immunity.Related researches have confirmed that actived NKT cells can inhabit tumor progress,but activated NKT cells commonly become anergic for some unknown reasons after a short time.This article reviews researches on the possible mechanisms and solutions to NKT cells anergy.

Role of NKG2 receptors-mediated natural killer cell activation in chronic HBV infection / 中华临床感染病杂志

The activation of NK cell , mediated by natural killer group 2 ( NKG2 ) family receptor , plays an important role in antiviral immune response and disease progression after hepatitis B virus (HBV) infection.To explore the NKG2 receptors-mediated NK cell activation and its mechanism may be of value for anti-HBV targeting immune treatment .This article reviews the recent research progress on the role of NK cells and its NKG2 family receptors in immunity of chronic HBV infection and its mechanisms .

Percentage and functions of natural killer T cells in peripheral blood and bone marrow of children with severe aplas-tic anemia / 临床儿科杂志

Objective To analyze the percentage and functional changes of natural killer T (NKT) cells in peripheral blood and bone marrow of severe aplastic anemia (SAA) children before immunosuppressive therapy (IST) comparing to that of healthy children.Methods Ten children with severe aplastic anemia were included in the study and ten healthy children at the same age were selected as the control group. By lfow cytometry, the percentage of CD3+CD1d tetramer+ NKT cell in peripheral blood and bone marrow were detected from March 2014 to December 2014 in our hospital. Immune magnetic bead separation was used to isolate and purify iNKT cells .The puriifed iNKT cells were cultured in the OCH(50 ng/ml,100 ng/ml or 200 ng/ml)+rhIL-2+rhG-CSF culture systems. The ampliifcation of iNKT cells after cultured in different systems were calculated. Elispot method was used to analyze the spotting form cells (SFCs) of IFN-γ or IL-4 expressed by activated iNKT cells.Results The percentage of CD3+CD1d tetramer+ NKT cells in peripheral blood of SAA group(0.72±0.03)% was signiifcantly lower than that of the control group(0.92±0.02)%(P=0.000). The percentage of CD3+CD1d tetramer+ NKT cells in bone marrow of SAA group(0.82±0.02)% was signiifcantly lower than that of the control group(1.05±0.05)%(P=0.000).In vitro iNKT cell ampliif-cation ability of bone marrow in SAA group was signiifcantly lower than the control group, and in medium concentration(50±6) and high concentration OCH group(52±6), the ampliifcation ability was higher than that in low concentration OCH group(30±5) (P<0.05). The secretion of IFN-γ in the iNKT cells of SAA bone marrow was signiifcantly lower in medium concentration(33±3) and high concentration(35±3)OCH group than that of the low concentration(50±3)OCH group(P<0.01). The secretion of IL-4 in the iNKT cells of SAA bone marrow was signiifcantly higher in medium concentration(50±3)and high concentration(75±3) OCH group than that of the low concentration(33±3) OCH group(P<0.01).Conclusions The quantity and function of NKT cells from children with SAA are lower than that of the healthy children.In vitro, they had better ampliifcation ability and could improve IL-4/IFN-γ imbalance in medium concentration and high concentration OCH group than in low concentration OCH group.

Deficiencies of Circulating Mucosal-associated Invariant T Cells and Natural Killer T Cells in Patients with Acute Cholecystitis

Mucosal-associated invariant T (MAIT) cells and natural killer T (NKT) cells are known to play crucial roles in a variety of diseases, including autoimmunity, infectious diseases, and cancers. However, little is known about the roles of these invariant T cells in acute cholecystitis. The purposes of this study were to examine the levels of MAIT cells and NKT cells in patients with acute cholecystitis and to investigate potential relationships between clinical parameters and these cell levels. Thirty patients with pathologically proven acute cholecystitis and 47 age- and sex-matched healthy controls were enrolled. Disease grades were classified according to the revised Tokyo guidelines (TG13) for the severity assessment for acute cholecystitis. Levels of MAIT and NKT cells in peripheral blood were measured by flow cytometry. Circulating MAIT and NKT cell numbers were significantly lower in acute cholecystitis patients than in healthy controls, and these deficiencies in MAIT cells and NKT cell numbers were associated with aging in acute cholecystitis patients. Notably, a reduction in NKT cell numbers was found to be associated with severe TG13 grade, death, and high blood urea nitrogen levels. The study shows numerical deficiencies of circulating MAIT and NKT cells and age-related decline of these invariant T cells. In addition, NKT cell deficiency was associated with acute cholecystitis severity and outcome. These findings provide an information regarding the monitoring of these changes in circulating MAIT and NKT cell numbers during the course of acute cholecystitis and predicting prognosis.

Effect of fungus on expression of interferon-γ and interleukin-4 in natural killer cells / 中华临床感染病杂志

Objective To investigate the effect of fungus Aspergillus fumigatus and Candida albicans on the expression of endocellular interferon-γ (IFN-γ) and interleukin-4 (IL-4) in cytokineinduced natural killer (NK) cells.Methods NK cells were cultured with Aspergillusfumigatus or Candida albicans by non-contact or direct-contact methods with a ratio of NK cells to fungus of 10 ∶ 1.The expressions of IFN-γ and IL-4 in NK cells were evaluated by flow cytometry after co-cultured for 6 h.Analysis of variance or SNK-q test was used to compare the expressions of IFN-γ and IL-4 among different groups.Results The IFN-γexpression rates in NK cells with direct contacting to Aspergillus fumigatus hyphae,or to different morphotypes of Candida albicans were (20.12 ± 0.53) ％,(20.69 ± 0.34) ％ and (20.8 ±0.37)％ respectively,while IFN-γexpression in NK cells with indirect contacting to fumigatus hyphae,or to different morphotypes of Candida albicans were (21.40 ± 0.53) ％,(20.57 ± 1.09) ％ and (20.20 ±0.51) ％ respectively,and all were significantly higher than that in the blank group [(15.11 ± 2.60) ％,all P ＞ 0.05].The IFN-γ expression rates in the Aspergillus fumigatus spores direct and indirect contacting groups were (14.33 ± 0.98) ％ and (14.97 ± 1.53) ％,which were not of significant difference compared with the blank group (P ＞ 0.05).The IL-4 expression rates in NK cells with direct contacting to different morphotypes of Aspergillus fumigatus and Candida albicans were (1.25 ± 0.06) ％,(1.21 ± 0.03) ％,(1.22 ± 0.46) ％ and (1.26 ± 0.11) ％,while those in indirect contacting groups were (1.21 ± 0.06) ％,(1.25 ±0.04)％,(1.27 ±0.03)％ and (1.26 ±0.1)％,which were not of significant difference compared with the blank group [(1.23 ± 0.05) ％,all P ＞ 0.05].Conclusion Fungus stimuli can reduce the secretion of IFN-γ in NK cells,but have not significant influence on the secretion of IL-4.