Universal chimeric Fcγ receptor T cells with appropriate affinity for IgG1 antibody exhibit optimal antitumor efficacy

Developing universal CARs with improved flexible targeting and controllable activities is urgently needed. While several studies have suggested the potential of CD16a in tandem with monoclonal antibodies to construct universal CAR-T cells, the weak affinity between them is one of the limiting factors for efficacy. Herein, we systematically investigated the impact of Fcγ receptor (FcγR) affinity on CAR-T cells properties by constructing universal CARs using Fcγ receptors with different affinities for IgG1 antibodies, namely CD16a, CD32a, and CD64. We demonstrated that the activities of these universal CAR-T cells on tumor cells could be redirected and regulated by IgG1 antibodies. In xenografted mice, 64CAR chimeric Jurkat cells with the highest affinity showed significant antitumor effects in combination with herceptin in the HER2 low expression U251 MG model. However, in the CD20 high expression Raji model, 64CAR caused excessive activation of CAR-T cells, which resulted in cytokine release syndrome (CRS) and the decline of antitumor activity, and 32CAR with a moderate affinity brought the best efficacy. Our work extended the knowledge about FcγR-based universal CAR-T cells and suggested that only the FcγRCAR with an appropriate affinity can offer the optimal antitumor advantages of CAR-T cells.

Preliminary study of cytomegalovirus infection and its correlation with NK cell subsets after renal transplantation / 器官移植

Objective To explore the variation trend of natural killer (NK) cell subsets in the recipients infected with cytomegalovirus (CMV) after renal transplantation. Methods Clinical data of 92 renal transplant recipients were retrospectively analyzed. All recipients were divided into the CMV infection group (n=43), CMV infection recovery group (n=13), stable renal function group (n=15), rejection group (n=11) and other infection group (n=10). In addition, healthy adult volunteers were enrolled in the healthy control group (n=15). The proportion of NK cells in peripheral blood, the expression proportion and the mean fluorescence intensity (MFI) of CD226 and CD16 in NK cells were observed and statistically compared among different groups. Results The proportion of NK cells was 4.9% (2.2%, 11.5%) in the CMV infection group and 3.7% (2.3%, 6.5%) in the CMV infection recovery group, which were significantly lower than those in the other groups (all P < 0.05). The expression proportion of CD226 and CD16 in NK cells in the CMV infection group was significantly lower compared with those in the healthy control group and stable renal function group(all P < 0.05). The expression proportion of CD226 and CD16 in NK cells in the CMV infection recovery group was remarkably higher than those in the CMV infection group (both P < 0.05). The MFI of CD226 and CD16 in the CMV infection group was significantly lower than those in the healthy control group (both P < 0.05). The MFI of CD226 and CD16 in the CMV infection recovery group was significantly higher than those in the CMV infection group (both P < 0.05). Conclusions The expression proportion and MFI of CD226 and CD16 in NK cells are down-regulated in CMV infection period, whereas up-regulated during the CMV infection recovery period, prompting that CD226 and CD16 expressed by NK cells are intimately correlated with the course of CMV infection.

Onset of Coronary Heart Disease is Associated with HCMV Infection and Increased CD14 CD16 Monocytes in a Population of Weifang, China / 生物医学与环境科学（英文）

Objective@#To investigate the relationship between human cytomegalovirus (HCMV) infection and peripheral blood CD14 CD16 monocytes in the pathogenesis of coronary heart disease (CHD), and to elucidate the mechanism of pathogenesis in CHD by analyzing the correlation between infection, inflammation, and CHD, to provide a basis for the prevention, evaluation, and treatment of the disease.@*Methods@#In total, 192 patients with CHD were divided into three groups: latent CHD, angina pectoris, and myocardial infarction. HCMV-IgM and -IgG antibodies were assessed using ELISA; CD14 CD16 monocytes were counted using a five-type automated hematology analyzer; mononuclear cells were assessed using fluorescence-activated cell sorting; and an automatic biochemical analyzer was used to measure the levels of triglyceride, cholesterol, high- and low-density lipoprotein cholesterols, lipoprotein, hs-CRp and Hcy.@*Results@#The positive rates of HCMV-IgM and -IgG were significantly higher in the CHD groups than in the control group. HCMV infection affects lipid metabolism to promote immune and inflammatory responses.@*Conclusion@#HCMV infection has a specific correlation with the occurrence and development of CHD. The expression of CD14 CD16 mononuclear cells in the CHD group was increased accordingly and correlated with acute HCMV infection. Thus, HCMV antibody as well as peripheral blood CD14 CD16 mononuclear cells can be used to monitor the occurrence and development of CHD.

Cytotoxic activity and subset populations of peripheral blood natural killer cells in patients with chronic pain

BACKGROUND: Chronic pain reportedly exerts complex effects on immune function. Natural killer (NK) cells are lymphocytes that play a critical role in cellular and innate immunity. This study examined changes in the subset populations and cytotoxic activity of peripheral blood NK cells in patients with chronic pain. METHODS: Thirty patients with chronic moderate-to-severe pain (group P) and age-matched pain-free subjects (group NoP) were enrolled. Peripheral whole blood was analyzed for the percentage and expression of NK cell surface markers (CD56 and CD16) by flow cytometry. Cytotoxic activity was assayed by evaluating CD69 expression on CD3−/CD56+NK cells. RESULTS: The percentage of NK cells among total lymphocytes was not significantly different between groups P and NoP (16.3 ± 9.3 vs. 20.2 ± 10.5%). Likewise, the percentages of two major NK cell subsets, CD56bright and CD56dim, were also not significantly different between the two groups. However, the percentage of CD56bright/CD16+ subset, was slightly but significantly increased in group P (1.0 ± 0.9%; P < 0.01) compared with group NoP (0.5 ± 0.6%). The cytotoxicity of NK cells was not different between the two groups, showing similar CD69 expression (P vs. NoP = 29.2 ± 15.2 vs. 32.0 ± 15.0%). These findings were not influenced by pain intensity, opioid use, or disease causing pain in group P. CONCLUSIONS: NK cell cytotoxic activity and major subset populations, with the exception of an increased percentage of the CD56bright/CD16+ subset, are not significantly altered in patients with chronic severe pain.

Clinical significances of the CD14bright CD16bright cell subset in peripheral blood of patients with gastric cancer / 中华微生物学和免疫学杂志

Objective To study the clinical significances of CD14bright CD16bright cell subset in pe-ripheral blood of patients with gastric cancer (GC). Methods The CD14bright CD16bright cells in peripheral blood samples collected from 124 patients with gastric cancer ( GC), 130 patients with chronic gastritis (CG) and 80 normal healthy controls (HC) were measured by using flow cytometry. Differences in the CD14bright CD16bright cells between different groups were analyzed with the Mann-Whitney U test. The feasibili-ty of using CD14bright CD16bright cells as a potential biomarker for differentiating GC patients from CG was as-sessed by using the receiver operating characteristic ( ROC) curve analysis. Correlations between the CD14bright CD16bright cells and clinicopathologic parameters of GC were analyzed with multivariate correlation analysis. Results The percentages of CD14bright CD16bright cells in peripheral blood samples and in CD14bright monomuclear cells collected from the patients with GC [median: 0. 38% (0. 23% -0. 52% ) and 6. 61%(4. 23% -9. 56% )] were significantly higher than those of the CG and HC groups [ median: 0. 11%(0. 07% -0. 15% ) and 5. 08% (3. 35% -6. 42% ); median: 0. 05% (0. 03% -0. 07% ) and 5. 09%(4. 20% -7. 40% )] (P<0. 01). The area under the ROC curve for CD14bright CD16bright cells in the peripher-al blood was 0. 934 (95% CI: 0. 900-0. 968) indicating that the value of CD14bright CD16bright cells in the di-agnosis of GC was much higher than that of alpha fetoprotein (AFP), cacino-embryonic antigen (CEA) and carbohydrate antigen CA199. The area under the ROC curve for combined multi-markers by using logistic model (CD14bright CD16bright cell subset and serum tumor markers) was 0. 947 (95% CI: 0. 920-0. 973). The CD14bright CD16bright cells were closely associated with lymphocyte cells ( P < 0. 01). Conclusion The CD14bright CD16bright cells were dramatically increased in the peripheral blood of patients with gastric cancer, which could be used as a biomarker in the diagnosis of gastric cancer.

Flow Cytometric Assays for Lymphocyte Subset Enumeration: CD45 is Inevitable for Lymphocyte Gating and CD16 is Essential for NK Cells

BACKGROUND: Clinical and Laboratory Standards Institute (CLSI) guidelines (H42-A2) recommend the "CD45/SSC" gating method for assays on lymphocyte subset enumeration and CD16 exclusion for assays enumerating NK cells. In contrast, the Flow Cytometry Checklist (06/17/2010) of the College of American Pathology does not recommend a specific lymphocyte gating method, but recommends the correction of lymphocyte subset results for lymphocyte gate purity. METHODS: We compared lymphocyte subset results of EDTA-treated blood from 102 patients with various diseases and 12 normal controls, using 3 lymphocyte gating methods (CD45/SSC, FSC/SSC, and lymphocyte gate purity correction after FSC/SSC gating), and assessed the proportion of CD56-/CD16+ NK cells within the total NK cell population. RESULTS: Lymphocyte gate purity increased as the percentage of lymphocytes increased. However, lymphocyte subsets that consistently showed high lymphocyte gate purity could not be identified. The purity of the T cell population differed significantly depending on the gating method used: CD45/SSC vs. FSC/SSC, P=0.027; CD45/SSC vs. gate purity correction after FSC/SSC, P=0.002. However, the lymphocyte gate purity correction after FSC/SSC gating did not significantly improve the accuracy of the lymphocyte subset enumeration assay using FSC/SSC gating. The subset of CD56-CD16+ NK cells, constituted an average of 17.1% of total NK cells. Patients had higher proportions of CD56-CD16+ NK cells (13.1-25.5%) than did the normal controls (9.52%). CONCLUSIONS: In flow cytometric assays to evaluate lymphocytic subsets, the CD45 is inevitable for lymphocyte gating, whereas the measurement of CD16 is essential for the evaluation of NK cell proportions.

Efecto de la proteína E2 del virus de hepatitis C sobre la activación de células NK / Effect of E2 protein of hepatitis C virus on the activation of NK cells

En estudios previos, se ha descrito un disminución de la activación y actividad citotóxica de las células NK en los pacientes infectados con hepatitis C; sin embargo, se desconoce el mecanismo por el cual éste fenómeno ocurre. En el presente reporte se estudió el efecto de la proteína E2 de la envoltura del virus o de la estimulación de su receptor con el anticuerpo anti-CD81 sobre la fosforilación de tirosinas, serinas, las enzimas: proteína quinasa C y fosfoinositol 3 quinasa, el factor de transcrición Nfkb y el intercambiador de nueclotidos VAV de células NK de controles normales estimulados con anti-CD16. Ambos, la proteína E2 y anti-CD81, combinado o por separado inducen una disminución de la fosforilación de tirosinas y serinas, así como una marcada disminución de la fosforilación de PKC, NFkB, PI3K y en menor grado VAV. Se concluye que la proteína E2 sola y en conjunto con anti-CD81 inducen señales inhibitorias responsables de la disminución en la activación de las células NK de pacientes infectados por el VHC y que éste fenómeno puede ser responsable de la cronicidad que se reporta en dicha enfermedad

The decrease in NK cell activation and cytotoxic activity in patients infected with hepatitis C virus has been described; however, the mechanism by whcih this phenomenon occurs is not known. In the present report, the effect of the E2 protein of the virus envelope or the stimulation of its receptor CD81 with the antibody anti-CD81 on the phosphorylation of tyrosines, serine, the enzymes protein kinase C, phosphoinositol kinase 3 (PI3K), the transcription factor NfkB and the nucleotide exchange protein VAV was assessed in NK cells from normal controls stimulated with anti-CD16. Both the protein E2 and anti-CD81 by themselves or combined, generated a decrease in tyrosine, serine, and a marked decrease in the phosphorylation of PKC, NfkB, PI3k and in less extent in VAV. It is concluded that the E2 protein alone and combined with anti-CD81 induce inhibitory signals responseible for the decrease in the activation of NK cells of infected HCV patients and it could be responsible for the chronicity observed in this disease

Efecto comparado de dosis inmunoestimulantes de fracciones ribosomales y proteoglicanos de membrana sobre la activación de subpoblaciones de células mononucleadas humanas de sangre periférica / Compared effect of immunostimulatory dose of ribosomal fractions and membrane proteoglycans on the activation of human mononuclear cell subsets from peripheral blood

Antecedentes. La población mexicana presenta una alta prevalencia de infecciones recurrentes de vías respiratorias altas. Objetivos. Comparar el efecto de dosis inmunoestimulantes de ribosomas bacterianos y proteoglicanos de membrana Ribovac® sobre células mononucleadas. Material y métodos. La expresión de IL-6 de células mononucleadas en cultivo, se midió a concentraciones y tiempos variables por la técnica de ELISA, mientras que el efecto de Ribovac® en poblaciones de células mononucleadas fue analizado por citometría de flujo. Resultados. Ribovac® tiene un efecto dependiente de dosis y tiempo de exposición sobre la expresión de IL-6 por células mononucleadas; las concentraciones de IL-6 fueron máximas a las 6 horas de tratamiento con Ribovac®. La expresión de CD3+ fue mayor cuando las células mononucleadas se trataron con 125 µg/ml por 72 horas (p=0,010) respecto a aquellas que se trataron a mitad de esa concentración en igual tiempo, a diferencia de la expresión de CD19, que fue mayor en células mononucleadas tratadas con 62,5 µg/ml por 72 horas que en aquellas tratadas con 125 µg/ml por 72 horas (p=0,021). No se encontraron disminuciones estadísticamente significativas en el número de células CD16+CD56+ ni en la coexpresión de los marcadores CD45 y CD19 cuando se compararon tanto tiempos de administración como concentraciones de Ribovac®. Conclusiones. La expansión de poblaciones linfoides y la maduración de éstas a fenotipos con mayor capacidad efectora son efectos inducibles y deseables de Ribovac®, que deben tenerse en cuenta al decidir su tiempo e intervalos de administración.(AU)

Background. The Mexican population has a high prevalence of recurrent infections of upper respiratory tract. Objective. To compare the effect of immunostimulatory dose of bacterial ribosomes and membrane proteoglycan Ribovac® on mononuclear cells. Methods. The expression of interleukin 6 from mononuclear cells in culture was measured at varying concentrations and times by ELISA, while the effect of R in populations of mononuclear cells was analyzed by flow cytometry. Results. Ribovac® has an dose-dependent and exposure time effect on the expression of IL-6 by mononuclear cells, concentrations of IL-6 were maximal at 6 hours of treatment with Ribovac®. The expression of CD3+ was higher when mononuclear cells weretreated with 125 µg/ml for 72 hours (p=0,010) than those who were treated to half that concentration in the same time, unlike the expression of CD19 which was higher in mononuclear cells treated with 62,5 µg/ml for 72 hours than those that were treated with 125 µg/ml for 72 hours (p=0,021). There were no statistically significance in the decrease in the number of CD16+CD56+ cells and in the coexpression of CD45 and CD19 markers neither; when comparing both times of administration and evaluated concentrations of Ribovac®. Conclusions. The lymphoid population expansion and their maturation to better effector phenotypes effector are inducible and desirable effects of Ribovac® and these important when deciding the time and intervals of administration.(AU)

A study of the effect of ligustrum quihoui carr (LQC)on the immune function of B lymphocytes and NK iymphocytes in mouse with impaired immune function / 国际中医中药杂志

ObjectiveTo explore the effect and mechanism of LQC on the immune function of B lymphocytes and NK lymphocytes in mouse with impaired immune function. Methods Mice with impaired immune function led by Cyclophosphamide (Cy)were used as experimental animmals, and divided into five groups randomly, twelve mice in every group, which is NS control group, Cy control group, Cy+ LQC minor and major dose group, and Cy + LQC decoction group. NS control group was given NS 0.2 ml/d subcutaneously 10 d, and the rest groups were given Cy 0.8 ml/d subcutaneously (20 mg/kg · d-1) 10d, to establish a mouse model of immune dysfunction. Since the 11th day each group was given NS, Cy, LQC minor dose, major dose and fructus ligustri lucidi decoctoin. Mice of each group were killed after 7 days. The percentage of B lymphocytes (CD3 - CD19 + ) and NK lymphocytes (CD3 - CD 16 + CD56 + ) in the peripheral blood of the experimental mice were detected by flow cytometer. ResultsIn comparison with LQC major dose group [ (20.44± 1.78)and(19.12± 1.70) ], fructus ligustri lucidi group[ (19.90± 1.42) and (20.17± 1.66) ], CD3-CD19+and CD3-CD16+CD56+ cells of LQC minor dose group[ (11.54±0.98) and (12.46±0.08)]were decreased significantly (P＜0.01), which were increased significantly compared with Cy group[ (4.53± 1.70) and (5.03 ±1.22) ] (P＜ 0.01), but they had no significant difference with NS [ ( 11.84 ± 0.99) and ( 12.90± 0.28) ] (P ＞ 0.05).In comparison with Cy group and NS group, CD3-CD19+ and CD3-CD16+CD56+ cells of LQC major dose group were increased significantly (P＜0.01), which had no significant difference with fructus lignstri lucidi group (P＞0.05). Conclusion The immune functions of the mice with impaired immune function were improved by LQC, it also could increase the quantity ofCD3-CD19+ cell and CD3-CD16+CD56+ cell. The dose of LQC was positively correlated with the modulation effect of LQC on the immune function.

Ausencia del receptor CD16b en neutrófilos / Neutrophils without CD16b receptors

Se presentan dos pacientes (mujeres de 41 y 15 años de edad) con ausencia del receptor para el fragmento Fc de IgG, CD16b en neutrófilos (fenotipo "null"). El caso 1 fue referida al laboratorio con diagnóstico de hemoglobinuria nocturna paroxística y el caso 2) con diagnóstico presuntivo de neutropenia inmune. En ambos casos se comprobó por citometría de flujo la ausencia de expresión de CD16b, sin deficiencias en la expresión de otras moléculas del sistema de alloantígenos propios de neutrófilos ni defectos en el anclaje a membrana por glicosil fosfatidil inositol (GPI). Las manifestaciones clínicas en ambas pacientes: anemia en el caso 1 y leucopenia en el caso 2 no pueden ser atribuidas exclusivamente a la carencia de CD16b, ya que otros receptores para Fc de IgG (CD32 y CD64) podrían suplir la función de CD16b. Sin embargo, es importante tener en cuenta esta rara deficiencia (b y neutropenia isoinmune natal transitoria en niños nacidos de mujeres con fenotipo "null".

Occurrence of the rare CD16b deficiency ("null" phenotype) in neutrophils from two female patients (41 and 15 years old) is reported. The first case was referred with a diagnosis of anemia related to paroxistic nocturnal hemoglobinuria and the second case, with presumptive diagnosis of immune neutropenia. In both cases, absence of CD16b expression was determined by flow cytometry without deficiencies of other neutrophil alloantigens or defects of membrane anchorage through glycosil phosphatydil inositol (GPI) linkage. Clinical manifestations in both patients could not be attributed exclusively to the absence of CD16b, as other receptors for the IgG Fc fragment (CD32 and CD64) could compensate this deficiency that occurs in < 1% of the caucasic population. Nevertheless, it is important to take this rare deficiency into account in order to prevent isoantibody formation after eventual blood transfusions, or transient neonatal immune neutropenia in children born to women with the "null" phenotype.

CD56/CD16 Expression on Mononuclear Cells and Concentration of Serum TNF-alpha in Recurrent Spontaneous Abortion / 대한진단검사의학회지

BACKGROUND: Recurrent spontaneous abortion (RSA) is defined as the occurrence of three or more consecutive spontaneous abortion before 20 gestational weeks. But, 40-50% of RSA still remain "unex-plained". Cytokines seem to play a critical role in the pathogenesis of unexplained RSA, and Th1 cytokines have been shown to exert deleterious effects on pregnancy. NK cytotoxicity has been reported to be predictive of subsequent abortion in women who had unexplained recurrent abortions. The aim of this study was to investigate immunophenotypic characteristics of peripheral blood mononu-clear cells and evaluate Th1 cytokine (TNF-alpha) production in women with RSA. METHODS: The study group comprised 93 women with RSA, and the control group consisted of 40 healthy pregnant women. The population of CD56/CD16 cells was observed by using a two-color scattergram in FACScan (Becton Dickinson, San Jose CA, USA). Concentration of TNF-alpha was measured by an enzyme-linked immunoabsorbant assay (ELISA) using commercial kits (NEOGEN corporation, Lexington KY, USA). RESULTS: The percentage of CD56+/CD16-cells were significantly higher (P<0.05) in the patients with RSA (13.40+/-7.95%) than in the pregnant control group (9.12+/-3.93%). We observed a significantly higher level of TNF-alpha (medians: 85.59+/-8.29 pg/mL versus 44.80+/-9.78 pg/mL; P<0.05) in RSA women compared to controls. CONCLUSIONS: This study indicates that an increased proportion of CD56+/CD16-mononuclear cells and increased level of serum TNF-alpha are related to RSA. Thus, the two factors could be used as an indicator of subsequent successful implantation and maintenance of gestation.

Human Activated Lymphocyte Treated with Anti-CD3, CD16, CD56 Monoclonal Antibody and IL-2

BACKGROUND: Throughtout the last three decades, the therapy of leukemias and lymphoma has set the stage for curative cancer therapy in systemic malignant disease. This was the result of an integrated work of basic reaserch and clinical investigators leading to more aggressive albeit tolerable protocol of chemotherapy and radiotherapy. High dose therapy marks the most elaborated strategies in this field today. However, intensification of conventional therapeutic modalities as mentioned has to be based on new approaches and the exploration of new antineoplastic mechanisms. This insight has resulted in immune therapy of cancer. Among the cells of the immune system, natural killer (NK) cells and T cells are of major interest for the development of therapeutic strategies. METHODS: Cytotoxicity to target cells was measured by LDH release method, Characterization of activated lymphocyte was measured by Flow cytometry analysis. Anti-CD3, 16, 56 monoclonal antibody and IL-2 were used for the activation of NK and T cell. The analysis of effect of activated lymphocyte, in vivo, were used by Balb/c nude mouse. RESULTS AND CONCLUSION: Cytotoxicity to K562 cells was significantly higher in the mixture group of NK and T cells than that of a group of activating T cells. The survivors and the rate of reduction of size of tumor craft of nude mouse group treatment with activated lymphocyte was higher than that of the group without treatment with activated lymphocyte. Therefore, this results are suggested that the activated lymphocytes by anti-CD3, CD16 and CD56 can reduce the malignancy effect of lymphoma.

Significance of Expression of CD_(14)~+ CD_(16)~+ on Peripheral Monocytes in Children with Kawasaki Disease / 实用儿科临床杂志

Objective To observe the significance of expressions of CD14+CD16+ on peripheral monocytes in children with Kawasaki di-sease (KD).Methods The expression of CD14+ and CD14+CD16+ monocytes in 16 children with KD (1-11 years old) were analyzed by flow cytomety both pre-treatment and post-treatment.And the percentages of CD14+CD16+ monocytes among CD14+ monocytes were calculated.Sixteen healthy children (10 months -10 years old) were served as normal control group.Statistical analysis was performed using t test.Results The levels of CD14+ monocytes,percentage of CD14+CD16+ monocytes among CD14+ monocytes and CD14+CD16+ monocytes in children with KD during acute phase (n=16) were (1.03?0.58)?109 L-1,(12.53?5.31)% and(1.20?0.79)?108 L-1.They were significantly higher than those in the normal controls[(0.57?0.21)?109 L-1,(3.86?1.84)% and (0.21?0.10)?108 L-1](Pa0.05).And the expressive levels remained high when the patient recurred.Conclusions The expressive levels of CD14+CD16+ monocytes increase in children with KD.And they change when the patient's clinical condition change.

Influences of two different HLA-B antigens on receptors expression of NK cells from peripheral blood lymphocytes / 中国免疫学杂志

Objective:To investigate influences of two different HLA-B antigens expressed on K562 cells on receptors expression of NK cells from peripheral blood lymphocytes.Methods:Studied the alteration of the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells before and after PBMC interaction with K562 cells for 24 hours,and also compared the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells after PBMC interaction with two different kind of K562 cells transfected with HLA-B39 and HLA-B51 respectively.Results:After PBMCs were incubated with K562 cells for 24 hours,the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells were both increased.However,after PBMCs were incubated with K562-HLA-B51 cells for 24 hours,the percentage of KIR3DL1+ cells and the percentage of CD16+CD56+ cells were both decreased in comparison with that interaction with K562-HLA-B39 cells.Conclusion:CD16 up-regulation was associated with an up-regulation of inhibitory receptors(KIR3DL1).The interaction between HLA-Bw4 and KIR3DL1 would down-regulate the expression of KIR3DL1.In addition,KIR3DL1 down-regulation was associated with down-regulation of activating receptors(CD16).

CD14/CD16 Subpopulations of Circulating Monocytes in Type 2 Diabetes Mellitus Patients with/without Coronary Artery Disease / 대한임상병리학회지

BACKGROUND: Coronary artery disease (CAD) is one of the most important causes of mortality and morbidity among patients with Type 2 Diabetes Mellitus (DM). Monocytes play a major role in the development of atherosclerotic lesions. The majority of circulating monocytes express high levels of the lipopolysaccharide receptor antigen (CD14) and low of the immunoglobulin Fc receptor III (CD16). Changes in the phenotype of circulating monocytes have been reported in patients with type 2 DM and CAD. The purpose of this study is to characterize the circulating blood monocyte subpopulations as potential cellular markers of systemic immunological abnormalities in CAD and DM and to evaluate the relationship among other independent risk factors. METHODS: Two-color immunofluorescence and flow cytometry was employed for evaluation of the monocyte subpopulations. CRP, HbA1c and lipid profile in patients with CAD (n=36) were also compared with those in a group without CAD (n=40) and healthy nondiabetic individuals (n=56). RESULTS: The CD14+(dim)/CD16+ (P<0.001) and CD14++/CD16- (P=0.011) subpopulations were significantly elevated in both the Type 2 DM patients' groups, with and without CAD, and compared with normal controls; and further, there were no significant differences between the diabetic groups. There was no correlation of the CD14+(dim)/CD16+ monocytes to any clinical parameter except for the number of CD14++/CD16-, which were positively correlated with the serum HbA1c (r=0.705, P<0.001). CONCLUSTIONS: In conclusion, the circulating blood monocyte subpopulations may not be the specific markers of atherogenesis in DM patients; however, these results suggest that they may play a role in systemic immunologic abnormalities in DM.

Expression and function of CD226 with CD16 on NK subsets / 中国免疫学杂志

Objective:To investigate the expression and function of CD226 on NK s ubsets and its coexpression with other activation receptor and inhibition recept or on NK cells. Methods:The expression of CD226 on CD56 bright and CD56 dim NK subsets and coe xpress ion with CD16 and NKG2A in PBMC and MLC in the presence or absence of IL-2 or IL-15 were detected by double fluorescent staining and flow cytometry analysis. The level of IFN-? in the supernatants of PBMC culture and MLC treated with o r without IL-2 or IL-15 were evaluated by ELISA. 51Cr release assay w as employed to measure the specific lysis of NK cells killing target K562 cells. Results:CD226 was mainly expressed on CD56 dim NK subsets in PBMC. When s imulated by IL-2, CD226 expression was shifted to CD56 bright NK subsets, while IL-15 increased the subpopulation of NKG2A+CD226+double positive cell s. In MLC-generated NK cells, CD226 was mainly expressed on CD56 dim NK su bsets, and also shifted to CD56 bright NK subsets in the addition of IL-15 . Furt hermore, the percentage of CD16+CD226+和NKG2A+CD226+ subsets were increa sed when stimulated by IL-2 or IL-15. There was great increase in IFN-? lev el in the supernatants of PBMC culture in the presence of IL-2 or IL-15, but no difference in the supernatants of MLC treated with or without two cytokine s. Moreover, the cytotoxicity of NK cells in PBMC and MLC were greatly enhanced by IL-2 or IL-15. Conclusion:CD226 is mainly expressed on CD56 bright NK subsets in IL-2 or IL-1 5 activated NK cells, and is coexpressed with CD16 and NKG2A preferentiatly, whi ch maybe involve in the modulation of cytotoxicity of NK cells based on the balance of coexpressed activation and inhibition receptors. [

Selective inhibition of polymorphonuclear leukocytes by immunosuppressive concentration of prostaglandin E2

Prostaglandin E2(PGE2) has been implicated as an immunosuppressive agent and plasma levels of PGE2 are elevated in patients sustaining thermal injury. We examined the effect of 10(-7) M prostaglandin E2(PGE2) on human polymorphonuclear leukocytes (PMN) to determine whether it directly inhibits stimulated responses of these cells. At this concentration, PGE2 alone was incapable of stimulating PMN intracellular hydrogen peroxide production (indirectly assayed by fluorescence of 2',7'ichlorofluorescin) or expression of the PMN CD11b/CD16 surface glycoproteins. PMN incubated in the presence of the soluble stimul phorbol myristate acetate(PMA, 100 ng/ml) or recombinant human C5a(rHC5a, 10(-8) M) generated significant amounts of hydrogen peroxide, increased their CD11b expression and decreased their CD16 expression. Pre-incubation of cells with PGE2 caused significant inhibition of all the observed changes stimulated by rHC5a. In contrast, events stimulated by PMA were not affected by preincubation of cells with PGE2. We conclude that PGE2, in concentrations identical to those found in the plasma of patients with burn injuries, is capable of selectively inhibiting some stimulated events and phenotypic expression of PMN in vitro study.

Polymorphonuclear leukocyte functions enhanced by chemotaxis

Human polymorphonuclear leukocytes (PMN) migrate into tissues in response to chemoattractants, yet it is not known whether this process alters the functional capabilities of the PMN. Using recombinant human interleukin-8 (rHIL-8, 100 ng/ml) as a stimulus, we compared a population of PMN that migrated through a polyvinylpyrrolidone-coated polycarbonate filter containing 8.0 microns diameter pores with PMN stimulated in suspension. PMN were analyzed by flow cytometry according to functional and phenotypic criteria. CD11b/CD16 expression was unaltered by chemotaxis. In contrast, chemotaxis enhanced phagocytosis of E. coli, independent of opsonization with IgG. Similarly, chemotaxis increased baseline hydrogen peroxide production. We conclude that the chemotactic motion of PMN "primes" the cell for increased oxidative burst activity and augments the ability of PMN to ingest bacteria. This increased functional capability is distinct from rHIL-8 stimulation and appears to be independent of complement-and Fc-receptor expression.

Influence of Dieda Paste on Expressions of CD16/32 and CD80 of Macrophages in Mouse Model with Traumatic Stasis / 中国药房

OBJECTIVE:To study the influence of Dieda paste application on expressions of CD16/32and CD80of mouse macrophages in traumatic condition.METHODS:The mice were divided into3groups:treatment group,model group and normal group.Using Rene method,7time-periods were adopted.Purified mouse peritoneal macrophages were obtained and detected with flow cytometer.RESULTS:After48-hour treatment,the subgroups of positive CD16/32,positive CD80and neg?ative CD16/32changed significantly.CONCLUSION:Topical application of Dieda paste could regulate the expressions of CD 16/32and CD80to normal levels.