Fcγ receptors on aging neutrophils

Abstract Objective Neutrophils are key effector cells of the innate immune system. They recognize antigens through membrane receptors, which are expressed during their maturation and activation. Neutrophils express FcγRII (CD32), FcγRIII (CD16), and FcγRI (CD64) after being activated by different factors such as cytokines and bacterial products. These receptors are involved with phagocytosis of IgG-opsonized microbes and enhance defense mechanisms. Based on that, our study seeks to compare the expression of FcγRII, FcγRIII, FcγRI, and CD11b on neutrophils from elderly and young subjects and their expression after in vitro activation with cytokines and LPS. Methodology Neutrophils were isolated from human peripheral blood and from mice bone marrow by density gradient. After isolation, FCγRs expression was immediately analyzed by flow cytometry or after in vitro stimulation. Results In freshly isolated cells, the percentage of FcγRIIIb+ and CD11b+ neutrophils were higher in samples from young individuals; FcγRIIIa expression was more prominent on aged neutrophils; FcγRIA expression was similar in all samples analyzed. Exposure to CXCL8 and LPS resulted in a higher percentage of FcγRIa+ neutrophils on elderly individuals' samples but lower when compared with neutrophils from young donors. We observed that LPS caused an increase in FcγRIIa expression on aging human neutrophils. In contrast, FcγRIIIb expression in response to CXCL8 and LPS stimulation was not altered in the four groups. CD11b expression was lower in neutrophils from elderly individuals even in response to LPS and CXCL8. In mice, we observed differences only regarding CD11b expression, which was increased on aged neutrophils. LPS exposure caused an increase in all FcγRs. Conclusions Our results suggest that, in humans, the overall pattern of FcγR expression and integrin CD11b are altered during aging and immunosenescence might contribute to age-related infection.

Application of neutrophil CD64 on the diagnosis and treatment of infectious diseases / 中华检验医学杂志

The early diagnosis of serious infection is very important since the fatality rate is quite high.Traditional diagnostic methods of infection includes procalcitonin,leukocyte count,CRP and so on,of which the sensitivity and specificity is not able to achieve the early diagnosis.Recently,neutrophil CD64 has been widely concerned for the high sensitivity and specificity in early diagnosis of bacterial infection.Flow cytometry is applied to detect neutrophil CD64 in early diagnosis of infectious diseases including respiratory infection,septicemia,neonatal intensive infection,burn and postoperative infection.What's more,the sensitivity and specificity can be further improved if neutrophil CD64 was combined with other inflammatory markers.Thus,neutrophil CD64 detected by flowy cytometry plays an important role in the diagnosis,monitoring,prognosis,therapeutic effect evaluation of infectious diseases.

Cross-reactivity of tree shrew sera with various secondary antibodies:extensive application to tree shrew models of diseases / 中国组织工程研究

BACKGROUND:Tree shrew is a representative between insectivore and primates, has a high degree of evolution, is more inexpensive primates, has high use of medical biology, and has been attached by scholars. OBJECTIVE:To detect whether the commonly used secondary antibodies have immune response with tree shrew serum. METHODS:Western blot assay and enzyme linked immunosorbent assay were utilized to detect whether the tree shrew serum had cross-reacts with anti-rabbit, anti-goat, anti-human, anti-mouse, anti-rat, and anti-monkey secondary antibodies. RESULTS AND CONCLUSION:Western blot assay results indicated that tree shrew serums did not react with anti-rabbit, anti-goat, anti-human, anti-mouse, and anti-rat secondary antibodies and had cross reaction with anti-monkey secondary antibody. Enzyme linked immunosorbent assay results also indicated that tree shrew serums were cross-reactive with anti-monkey secondary antibody, but did not have cross-reactivity with the other secondary antibodies. Above data confirmed that the usual y soled secondary antibody cannot be used to immunoassay with tree shrews IgG. Only anti-monkey secondary antibody has cross-react with tree shrew serum. It is necessary to prepare anti-tree shrew IgG monoclonal and polyclonal antibodies. When no antibody is readily available at present, anti-monkey secondary antibody can be used to substitute detection, and can be widely applied in the study of tree shrew models of disease.

Value of neutrophil CD64 in diagnosis of sepsis in adult patients:a meta-analysis / 中华临床感染病杂志

Objective To evaluate the application of neutrophil CD 64 in diagnosis of sepsis in adult patients.Methods Literature retrieval from PubMed, EMBASE, ISI Web of Knowledge, Cochrane Library, CBM, CNKI, CQVIP, Wanfang Data from the establishment of database to the year 2015 was conducted to identify all studies on CD 64 in diagnosis of sepsis .The quality of the literature was evaluated with the quality assessment of diagnostic accuracy studies ( QUADAS).Meta-Disc 1.4 and STATA 12.0 were used for meta analysis . Fixed-effects or random-effects model was performed based on the heterogeneity.The sensitivity, specificity, positive likelihood ratio, negative likelihood ratio and diagnostic odds ratio were calculated .Summary receiver operating characteristic curves ( SROC ) and area under the curve (AUC) were used to evaluate the diagnostic performance of CD 64 for sepsis.Results A total of 24 studies involving 3 198 patients were included for systemic review .The sensitivity, specificity, positive likelihood ratio, negative likelihood ratio and diagnostic odds ratio of CD 64 for diagnosis of sepsis were 0.79 (95 %CI:0.77-0.81), 0.86 (95 %CI:0.84-0.88), 7.40 (95 %CI:5.02-10.91), 0.15 (95 %CI:0.10-0.22) and 60.07 (95%CI: 29.19-123.60), respectively.The area under SROC of CD64 in diagnosis of sepsis was 0.95, and the Q* value was 0.88.Conclusion CD64 can be used to diagnose sepsis in adult patients , but it needs to be further confirmed by large multicenter studies .

Clinical value of CD64 index in early diagnosis for bacterial infection in children / 国际检验医学杂志

Objective To investigate the diagnostic value of neutrophil CD64 expression for bacterial infection in children . Methods 168 children were divided into the infection group (n=133) and non-infection group(n=35) according to the results of bacterial culture .CD64 reagents from Beckman company and BD company were employed to detect CD 64 in neutrophils and lym-phocytes .Flow cytometry was used to assay their average fluorescence intensity and the CD 64 indexes were calculated .Receiver operator characteristic(ROC) curve was adopted to analyze the diagnostic performance of CD64 indexes .Results Average fluores-cence intensity of neutrophil CD64 detected by CD64 reagents from Beckman company and BD company were 46 .16 ± 29 .21 , 28 .11 ± 17 .90 ,respectively ,with statistical difference(P0 .05) .The CD64 index of children in infection group (7 .06 ± 4 .20) was higher than that in the non-infection group(2 .93 ± 0 .79)(P<0 .01) .When CD64 index cut-off point was 3 .37 ,the sensitivity and specificity for bacterial infection diagnosis of CD64 were 93 .2% and 82 .9% ,respectively .Conclusion CD64 index may be served as an effective indicator for early diagnosis of bacterial infection in children .

Early diagnosis of sepsis with C-reactive protein, serum amyloid A and CD64 in very low birth weight infants / 中华围产医学杂志

Objective To investigate the value of C-reactive protein (CRP),serum amyloid A (SAA) and CD64 in early diagnosis of sepsis in very low birth weight infants (VLBWI).Methods Fifty-four VLBWI suspected to sepsis and 20 VLBWI without infection in neonatal intensive care unit of the Second Hospital of Wenzhou Medical College from May 2010 to May 2012 were enrolled in this study.CRP,SAA and CD64 of sepsis group were measured at 0 and 24 hour after suspected bacterial infection; and those of control group were measured at corresponding age.CRP and SAA were detected by enzyme-linked immunosorbent assay,and CD64 was detected by flow cytometry.The difference between groups was compared by Mann-Whitney U test.Receiver operating characteristic curve was used to predict the sensitivity and specificity of the three biomarkers on sepsis.Results Fifty-four VLBWI were suspected with sepsis,and 37 patients were finally diagnosed.The levels of the three biomarkers in sepsis group were higher than those of control group not only at 0 h [CRP:13.3 mg/L(4.6-67.2 mg/L) vs 4.4 mg/L(1.6-11.2 mg/L),Z=-2.308; SAA:95.7 mg/L(4.5-265.9 mg/L) vs 7.3 mg/L(2.5-16.9 mg/L),Z=-2.425; CD64:7306 fluorescent antibody molecules/cell (2667-10 853 fluorescent antibody molecules/cell) vs 2502 fluorescent antibody molecules/cell (1839-3017 fluorescent antibody molecules/cell),Z=-3.704],but also at 24 h[CRP:35.4 mg/L (7.7 106.5 mg/L) vs 3.2 mg/L (1.1-7.8 mg/L),Z-5.501; SAA:359.3 mg/L (3.8-503.2 mg/L) vs 6.6 mg/L (3.0-12.7 mg/L),Z =-2.818; CD64:8304 fluorescent antibody molecules/cell (2819-11 758 fluorescent antibody molecules/cell) vs 2563 fluorescent antibody molecules/cell (1760-3154 fluorescent antibody molecules/cell),Z =-7.670],P＜0.05 respectively.The best cutoff value of CD64 was 2934 fluorescent antibody molecules/cell,with the sensitivity of 81.1％ at 0 h and 91.9％ at 24 h; and the specificity of 90.0％ at 0 h and 80.0％ at 24 h.Although SAA had similar sensitivity (0 h:83.8％; 24 h:86.5％) as CD64,its specificity was relatively low(0 h:65.0％; 24 h:55.0％).Both the sensitivity (0 h:62.2％; 24 h:70.3％) and specificity (0 h:70.0％; 24 h:70.0％) of CRP were low.Combination of the three infection biomarkers could increase the sensitivity (0 h:91.9％; 24 h:97.2％) and specificity (0 h:95.0 ％ ; 24 h:90.0％).Conclusions Combination of CRP,SAA and CD64 might improve the diagnostic accuracy of sepsis in VLBWI.

Expression of FcγRⅡb on peripheral B cells and its clinical significance in patients with primary Sj(o)gren's syndrome / 中华风湿病学杂志

Objective To investigate the expression of FcγRⅡb on peripheral B cells and its clinical significance in primary Sj(o)gren's syndrome (pSS).Methods FcγRⅡb expression on peripheral B cells from 19 pSS patients and 15 healthy controls was examined by flow cytometry.The levels of serum anti-SSA and SSB antibodies were determined using enzyme-linked immunosorbent assay (ELISA).Data were analyzed with t test,one-way ANOVA,SNK-q test and Pearson's correlation test.Results The percentage of memory CD19+CD27+ B cell subpopulation was significantly lower in pSS patients [ (20.8±2.7)％] when compared to normal controls [(37.8±2.2)％ ](t=-4.002,P＜0.01).The level of expression of FcγRⅡb in active pSS memory CD19+CD27+ B cells [ MFI(74±8)] was significantly reduced when compared to inactive [ MFI( 132±11)] and normal controls [ MFI (139±12)] (F=10.699,P＜0.01).The level of expression of FcγRⅡb on memory CD19+CD27+ B cells from pSS patients was inversely correlated with Sj(o)gren's syndrome disease activity index (SSDAI) (r=-0.744,P=0.0003 ).pSS patients with the serum anti-SSA/SSB antibodies positive group [ MFI(75+3),(48±7)] displayed a lower expression of FcγRⅡb on memory CD19+CD27+ B cell than in patients with the serum anti-SSA/SSB antibodies negative group [MFI( 122±11),(108±9)] (t=-4.336 and -3.776 respectively,the P value of both tests were less than 0.01).The level of expression of FcγRⅡb in the memory CD19+CD27+ B cells of patients with active pSS was inversely correlated with anti-SSA antibody titers (r=-0.685,P=0.014).Conclusion The expression of FcγRⅡb on peripheral memory B cells from active pSS patients is inversely correlated with SSDAI and is also inversely associated with anti-SSA antibody levels.Decreased expression of FcγRⅡb might play an important role in the pathogenesis of pSS.

Dermatose por IgA e IgG linear: relato de caso com boa resposta terapêutica à dapsona e ao micofenolato mofetil / Linear IgA/IgG bullous dermatosis: successful treatment with dapsone and mycophenolate mofetil

Relata-se o caso de paciente feminina, de 21 anos, com dermatose por IgA e IgG linear. Inicialmente, a resposta clínica foi favorável à dapsona. Após a interrupção desta medicação, por crise de anemia sintomática, precipitada por malária, houve piora da doença, apesar da utilização da prednisona e pulsoterapia com metilprednisolona. A reintrodução da dapsona, associada ao micofenolato mofetil, possibilitou o controle da enfermidade.

A 21-year-old female presenting linear IgA and IgG disease initially responded well to dapsone therapy. However, the treatment with dapsone was withdrawn due to severe anemia induced by malaria, which led to worsening of the clinical picture. Although prednisone and methylprednisolone were tried, the patient responded only to the association of dapsone and mycophenolate mofetil.

Dynamic observation on IgG and its subclasses and IgE in sera of mice by immunization with mixed recombinant of BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine of Echinococcus multilocularis / 中国地方病学杂志

Objective To dynamically observe changes of IgG, its subclasses and IgE in sera of mice by immunization with mixed recombinant of BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine of Echinococcus multilocularis (Era). Methods Forty Balb/c mice of 12-14 week old and 20-25 g weight were intranasally vaccinated by the vaccine, 4 mice were killed randomly by the weight on 0,2,4,6,8,10,12,14,16 and 18 weeks of immunization respectively, sera were gathered from the eyeball to measure IgG, its subclasses and IgE by routine ELISA. Results Levels of IgG, IgG2a and IgG2b in the sera of mice increased obviously on 2-18 weeks, reached the highest level on 10, 4 and 4 weeks respectively, the value was 0.095±0.033,0.022±0.001,0.023±0.003 respectively, as compared with the value on 0 week(0.030±0.013,0.012±0.004,0.013±0.004), the difference being statistically significant(q=2.95,4.87,2.81 respectively, P < 0.01 or P < 0.05); levels of IgG1, IgG3 and IgE in the sera of mice decreased remarkably on 2-18 weeks,came to the lowest level on 4,2,6 weeks respectively, the value was 0.031±0.004,0.136±0.002,0.114±0.002 respectively, as compared with the value on 0 week(0.192±0.007, 0.175±0.013,0.024±0.003), the difference being statistically significant (q =5.16,4.93,5.32 respectively, P < 0.01 or P < 0.05). Conclusion Helper T cell(TH) Ⅰ response is induced in mice by mixed recombinant of BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine on early immunization.

The expression of neutrophils FCγRⅡ/Ⅲ modulated by anti-neutrophil cytoplasmic antibodies and its clinical value / 中华风湿病学杂志

Objective The interaction between anti-neutrophil cytoplasmic antibaties (ANCA) and receptors at the Fc portion of immunoglobulins (FeγR) is central in the pathogenisis of primary systemic small vasculitis. The aim of this study is to investigate the role and clinical value of ANCA on the expression of neutrophils FCγRⅡ/Ⅲ (CD32/CD16). Methods ANCA IgG was prepared from the sera of patients with active We-gener's granulomatosis (WG) and microscopic polyangiitis (MPA). Neutrophils were isolated from the blood of healthy volunteers. The expression of CD32/CD16 on neutrophils was assessed by flow cytometry after stimulated by ANCA for 1 hour. We compared the expression of CD32/CD16 between 18 primary systemic small vaseulitis (PSV) patients and 35 healthy volunteers. Furthermore, the correlation was also be analyzed between the expression of CD32/CD16 and Birmingham vasculitis activity score (BVAS). Results The expression of CD16 was significandy elevated by ANCA (Mnx 67±23 vs 54±21, P<0.01 ). The expression of CD16 was higher in patients than in healthy volunteers (Mnx 62±12 vs 53±10, P<0.01), which was in correlation with BVAS (r=0.728 86, P<0.01). But no such correlation was found for CD32 . Conclusion ANCA may play a role in the pathogenesis of PSV by modulating the expression of the FCγR. Monitoring the expression of CD16 on neutrophils is helpful for the evaluation of PSV activity.

DETERMINATION OF LYMPHOCYTE SUBSETS IN THE PERIPHERAL BLOOD OF THE PATIENTS WITH ALLOGENCIC HAND TRANSPLANTATION / 解放军医学杂志

To investigate the changes in the activated T-lymphocyte CD3/HLA-DR and CD3/CD(16+56) populations in peripheral blood of the patients with allogeneic hand transplantation, lymphocytes from peripheral blood of the patients at different time points were immunologically labeled with dual color fluoresecent monoclonal antibodies CD3/CD(16+56) and CD3/HLA-DR, mono-color fluoresecent monoclonal antibody CD25. CD25, CD3/CD(16+56), and CD3/HLA-DR were determined with flow cytometry (FCM). The levels of activated T-lymphocyte (CD25 +,CD3 +/HLA-DR + ), silent T-lymphocyte [CD3 +/CD(16+56) -,CD3 +/HLA-DA - ] decreased significantly during the first week after transplantation and then increased gradually to the pre-operafive level. Nature killer cells [CD3 -/CD(16+56) +] increased significantly at the first day after transplantation, then decreased sharply and maintained a lower level. The results suggest that immunosuppressive agents have significantly effects on lymphocyte subsets in allogenaic hand transplanted patients, and dynamic determination of HLA-DR, CD3 /CD(16+56) could be valuable in immunomonitoring after allogeneichand transplantation.

Dynamic changes and significance of subsets of blood lymphocyte in 206 adult sev ere acute respiratory syndrome patients / 中华传染病杂志

Objective To study the changing of subsets of blood lymphocyte in adult SARS patients and its effect on the clinical features and prognosis. Methods According to the clinical characteristic diagnostic standards of SARS recommended by the Ministry of Health of China, 206 of hospita lized SARS patients were divided into 3 groups: Mild-Moderate group included 13 3 patients; severe group 50 patients and death group 23 patients, and cells coun t changes of CD4 +, CD8 +, CD19 + and CD16 +. Statistic analyses were perfor med to analyze the relationship of immune changes and clinical features and prognosis. Results The counts of CD4 +, CD8 +, CD19 + lymphocytes in mild-moderate group were h igher than severe group, while lowest in death group (P0.05), there were significanl y difference in CD4 +, CD8 +,CD19 + and CD16 + cell counts among three grou ps(P

Study of Fc?RⅢA-158F/V polymorphism in patients with rheumatoid arthritis / 中华风湿病学杂志

Objective The purpose of this study is to confirm the putative association between the Fc?RⅢA-158F/V allele and rheumatoid arthritis (RA) by genotyping the Fc?RⅢA-158F/V polymorphism in patients and controls. Methods One hundred and ninety-two RA patients were recruited and all patients fulfilled the American College of Rheumatology 1987 revised criteria for RA. The demographic data including age, sex and disease duration were collected. The 192 RA patients and 179 control subjects were genotyped for Fc?RⅢA-158F/V polymorphism with a reliable polymorphism assay, nested PCR to investigate whether the Fc?RⅢA-158F/V polymorphism was a risk factor for RA. Results The frequency of Fc?RⅢA-158V/V homozygous was higher in RA patients (OR=3.1, P

Polymorphism of Fc Gamma Receptor Ty peⅢin Han Patients with Systemic Lupus Erythematosus from Hunan Province / 中华皮肤科杂志

Objective To investigate the association betw een systemic lupus erythematosus(SLE)and polymorphismof Fc gamma receptor ty peⅢin Han patients fromHunan province.Methods Genotypes of Fc?RⅢa-158V/F were determined by polymerase chain reaction(PCR)and restriction fragment length polymorphism(RFLP)analysis in 65patients with SLE and 60normal controls.Results①It was found that the frequency of homozygous Fc?RⅢa-158F /F genotype was significantly h igher in patients with SLE than that i n controls(OR=2.23,? 2 =4.69,P=0.03).②The frequencies of both homozygous Fc?RⅢa-158F /F genotype and Fc?RⅢa-158F allele were significantly high er in patients with lupus nephritis c ompared with those in controls(OR=2.67,? 2 =5.36,P=0.02;OR=2.00,? 2 =4.91,P=0.03).Conclusions These results suggest that an abnorm al distribution of Fc?RⅢa-158V/F polymorphism is associated with SLE in the Hans of Hunan province,and the presence of Fc?RⅢa-158F allele is a risk factor for lupus nephritis.These findings support t he hypothesis of a genetic mechanism in the pathogenesis of SLE.[