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Persistent HBV infection alters the expression of receptors on the surface of innate and acquired immune cells, which may cause a variety of immune disorders and finally lead to immune escape and disease chronicity. Studies have shown that the upregulation of inhibitory receptors is the main cause of immune disorders in patients, and blocking inhibitory receptors can restore immune function to a certain extent. T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) is a new type of inhibitory receptor attracting much attention at present, and it is highly expressed in NK cells and T cells. It has been found that TIGIT plays an important role in chronic viral infection, and this article briefly reviews the research advances in the association between TIGIT and immune disorders in chronic HBV infection.
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Inflammatory reaction dominated by defense response will arise against infection and trauma. As an important proinflammatory cytokine, high mobility group box 1 (HMGB1) is widely expressed in all nuclear cells to mediate the inflammatory response. However, the biological functions of HMGB1 in inflammation vary depending on the type of HMGB1 protein modification and the localization in the cell. HMGB1 protein will be modified as acetylation of lysine residues, methylation of lysine residues, oxidation of cysteine residues, phosphorylation of serine residues, glycosylation of asparagine residues, adenosine diphosphate-ribosylation and lactylation of the protein in the nucleus, migrate from the nucleus to the cytoplasm, and release into the extracellular compartment. Extracellular HMGB1 can bind to receptors for advanced glycation end products (RAGE) and Toll-like receptors, activate cells and regulate inflammatory responses. The authors review the research progress in regulatory mechanism of HMGB1 in inflammation response from aspects of its post-translational modifications, releases, biological roles and binding receptors, hoping to provide theoretical basis for finding the targets of inflammation intervention.
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Objective@#To investigate the expression of inhibitory receptor TIGIT gene in peripheral NK cells of patients with rheumatoid arthritis (RA) and its clinical significance.@*Methods@#A case control study was conducted of 58 RA patients(30 patients with active RA disease, 28 patients with remission of RA) and 22 healthy controls (HC) in the department of rheumatology and immunology from Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine during December 2018 to June 2019, the related clinical data were collected. Flow cytometry was used to compare the expression of TIGIT gene on peripheral NK cells. The IFN-γ secretion level of cytokines in peripheral blood was detected by ELISA, and analyzed the correlations between TIGIT gene and disease activity and IFN-γ level. T-test or non-parametric test was used for comparison between the two groups, and Pearson correlation analysis was used for correlation between the two variables.@*Results@#Compared with HC group, the number of NK cells in RA disease group was reduced, which was (13.88±4.56) ×107 cells/L in RA disease group and (25.69± 2.48) ×107 cells/L in HC group (t=-2.036, P=0.041). The percentage of TIGIT gene expression in peripheral blood NK cells was not statistically different between RA disease group (39.73±9.37)% and HC group (45.64±9.91)% (t=-1.241, P=0.218). However, the average fluorescence intensity (MFI) of TIGIT gene expression was decreased, which was (7.21±2.03) in RA group and (9.01±3.29) in HC group (t=-2.947, P=0.004).MFI of TIGIT gene in NK cells of the disease active subgroup was (6.72±2.01), lower than that of the disease remission subgroup (8.75±2.64), (t=-3.316, P=0.002), and MFI of TIGIT gene in NK cells of the disease active subgroup was negatively correlated with DAS28 score (r2=0.649 6, P<0.000 1).The secretion level of IFN-γ cytokine in the RA disease group was (67.13±14.84) pg/ml, higher than that in the HC group (57.21±14.23) pg/ml (t=2.757, P=0.017), and the secretion level of IFN-γ cytokine in the disease active subgroup was negatively correlated with the MFI of TIGIT gene on NK cells (r2=0.662 2, P<0.000 1). Experimental results of peripheral blood mononuclear cell stimulation in the RA disease activity subgroup showed that the secretion level of cytokines IFN-γ was reduced after stimulation compared with that before stimulation (t=11.38, P<0.000 1).@*Conclusion@#The abnormal expression of TIGIT gene on peripheral NK cells are observed in patients with RA, which correlate with disease activity and IFN-γ secretion level.
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Objective To evaluate the relationship between the triggering receptor expressed on myeloid cells (TREM) and postoperative cognitive dysfunction (POCD) in elderly patients.Methods Eighty American Society of Anesthesiologists physical status Ⅰ-Ⅲ patients,aged 65-85 yr,weighing 50-80 kg,scheduled for elective total knee replacement under spinal-epidural anesthesia were enrolled in this study.Cerebrospinal fluid (CSF) was extracted after a catheter was successfully inserted into subarachnoid space.Blood samples from the cubital vein was collected before anesthesia induction (T0) and at 24 and 72 h after surgery (T1,2).The concentrations of TREM1 and TREM2 in CSF and plasma and tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in plasma were measured by enzyme-linked immunosorbent assay.The expression of TREM1,TREM2,IL-6 and TNF-α protein and mRNA in mononuclear ceils in peripheral blood was detected using real-time polymerase chain reaction.Neuropsychological test was performed in the the same time period at 1 day before surgery and 7 days after surgery,and the Z score was used to diagnose the development of POCD.The patients were divided into POCD group (P group) and non-POCD group (NP group) according to whether or not POCD happened after surgery.Results The incidence of POCD was 22％.Compared with group NP,the plasma TREM1 concentrations at T1,2 and plasma IL-6 and TNF-α concentrations at T2 were significantly increased,and the expression of TREM1 mRNA and TNF-α mRNA at T1,2 and IL-6 mRNA at T2 was up-regulated in group P (P＜0.05).There was no significant difference in plasma TREM2 concentrations at each time point between and within groups (P＞0.05).There was a higher consistency between plasma and CSF TREM1 concentrations (Cronbach's Alpha=0.784,P＜ 0.01) and a high consistency between plasma and CSF TREM2 (Cronbach's Alpha =0.935,P＜0.01).Conclusion Up-regulated expression of central and peripheral TREM1 is related to the development of POCD in elderly patients.
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Objective To evaluate the role of etomidate post-conditioning on mitochondrial permeability transition pore (mPTP) in the rat cortical neurons subjected to oxygen-glucose deprivation and restoration (OGD/R) and the relationship with Robo receptors.Methods The cortical neurons obtained from Sprague-Dawley rats (＜ 24 h after birth) were cultured in vitro and seeded in 6-well plates (2 ml/well).The neurons were divided into 4 groups (n=24 each) using a random number table:control group (group C),OGD/R group,etomidate post-conditioning group (group E),and etomidate post-conditioning + Robo receptor blocker group (group ER).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h.In E and ER groups,etomidate was added to the culture medium with the final concentration of 6 μmol/L immediately after onset of O2-glucose supply.In group ER,Robo blocker RoboN was added to the culture medium with the final concentration of 1 μg/ml at 6 h before O2-glucose deprivation.The neuronal apoptosis was detected using Hoechst/PI double staining,the viability of neurons was measured by MTT assay,and the amount of lactic dehydrogenase (LDH) released was measured using colorimetric method.The mitochondria were extracted,and mitochondrial permeability transition pore (mPTP) opening was detected.Results Compared with group C,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in OGD/R,E and ER groups (P＜0.05).Compared with group OGD/R,the apoptosis rate,amount of LDH released,and mPTP opening were significantly decreased,and the cell survival rate was increased in group E,and the apoptosis and amount of LDH released were significantly decreased,and the cell survival rate was increased in group ER (P＜0.05).Compared with group E,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in group ER (P＜0.05).Conclusion Etomidate post-conditioning mitigates OGD/R-induced damage to the cortical neurons through activating Robo receptors and inhibiting mPTP opening in rats.
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Objective To investigate the correlation of the expressions of advanced glycation end products(AGE) and the receptor for advanced glycation end products(RAGE) in serum and placenta with the pathogenesis of preeclampsia. Methods From December 2013 to June 2014, 32 women with severe preeclampsia who received cesarean section in the Affiliated Hospital of Qingdao University were recruited in the study, defined as the severe preeclampsia group. 30 healthy pregnant women who received cesarean section in the same hospital were recruited as the control group. ELISA was used to measure the maternal serum AGE, soluble receptor for advanced glycation end products (sRAGE) and tumor necrosis factor-α(TNF-α) in these women. Furthermore, ELISA was also used to measure AGE and TNF-α in the placenta. The localizations of AGE and RAGE protein in placentas were detected by immunohistochemical SP method. RAGE and TNF-α mRNA expression in placentas were measured by real-time quantitative PCR. AGE, RAGE and TNF-αprotein expression in placentas were measured by western blot, respectively. Results (1) The serum levels of AGE,sRAGE and TNF-αin the severe preeclampsia group were (538 ± 75),(367 ± 86) and (322 ± 40) ng/L,respectively. They were significantly higher than those in the control group[(454 ± 50), (286 ± 35) and (270 ± 35) ng/L, respectively](P0.05). (2) In the severe preeclampsia group, the levels of AGE and TNF-αin placentas were (500 ± 82) and (334 ± 57) ng/L, which were higher than those in the control group [(431 ± 74) and (263 ± 46) ng/L, respectively](P<0.05). The levels of AGE showed positive correlation with the levels of TNF-ɑ(r=0.406,P<0.05). (3)AGE and RAGE protein mainly located in the syncytiotrophoblasts, macrophages and vascular endothelial cells in the placentas of the two groups. AGE expressed mainly in the cytoplasm, and RAGE expressed in the cytoplasm and cell membranes.(4)RAGE and TNF-αmRNA expression in the placentas of the severe preeclampsia group were 12.6 ± 4.6 and 10.4 ± 2.4, which were significantly higher than those in the control group (0.9 ± 0.4 and 3.5 ± 0.9,P<0.01). (5) The expressions of AGE、RAGE and TNF-αprotein in placentas of the severe preeclampsia group were 0.68 ± 0.06, 0.82 ± 0.08 and 0.76 ± 0.08. All were significantly higher than those of the control group (0.46 ± 0.05,0.42 ± 0.09 and 0.52 ± 0.07;P<0.01). Conclusions The levels of AGE and RAGE in serum and placentas elevated in the severe preeclampsia group, and the expression of TNF-αalso elevated. These indicated that AGE and RAGE might be involved in the systemic inflammatory response and local inflammatory response in placentas, and then caused the preeclampsia.
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Objective To investigate the triggering receptor expressed on myeloid cells-1(TREM-1) of cord blood leukocytes in neonates and the transcription level of mRNA, and analyze its promoting function of inflammatory cytokine secretion. Methods During the period from September 2013 to March 2014, cord blood was collected from 20 term neonates at the time of birth, and peripheral blood was collected from 20 healthy adults. The expression of TREM-1 and TREM-1 mRNA on leukocytes was observed using flow cytometry and real-time reverse transeription-polymerase chain reaction, respectively. After the whole cord blood was stimulated by lipopolysaccharide (LPS) or LP17 plus LPS, the contents of interleukin (IL)-6, tumor necrosis factor-α (TNF-α), IL-8 and soluble TREM-1 (sTREM-1) in the supernatant were analyzed by enzyme linked immunosorbent assay. The statistical significance was determined using the one-way ANOVA test, t test, q test and Pearson correlation coefficient. Results The mean fluorescence intensity of TREM-1 on leukocytes of newborns was not different compared with healthy adults (P>0.05), while the percentage of TREM-1 positive on polymorphonuclear cells was lower than that of healthy adults [(82.3±7.1)% vs (98.6±4.8)%, P<0.05]. The level of TREM-1 mRNA in newborns was lower than in healthy adults (1.16±0.13 vs 1.63±0.24, t=7.714, P<0.01). The LPS treatment significantly increased sTREM-1 in newborn whole blood compared with the control treatment [(156.7±36.3) vs (34.6±6.1) pg/ml, t=13.623, P<0.01]. The concentration of IL-6, TNF-αand IL-8 decreased significantly when TREM-1 was blocked by LP17. In addition, the concentration of sTREM-1 showed a positive correlation with the levels of TNF-α(r=0.519, P<0.05), IL-6 (r=0.507, P<0.05) and IL-8 (r=0.538, P<0.05). Conclusions Healthy newborns exhibit expression of TREM-1 on monocytes similar to healthy adults, and most PMNs express TREM-1 at the newborn stage. Blocking the TREM-1 signal transduction pathway may reduce inflammatory responses of neonate leukocytes.
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Objective To investigate the role of S100 calcium binding protein A12 (S100A12) in the pathogenesis of preeclampsia. Methods Sixty patients with preeclampsia were recruited from March 2013 to December 2013 in the First Affiliated Hospital of Zhengzhou University. Among them, thirty cases were defined as the mild preeclampsia group and thirty cases were defined as the severe preeclampsia group. The other thirty healthy pregnant women were recruited in the healthy pregnant women group. The levels of S100A12 protein in maternal peripheral blood were detected by enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry of streptavidin peroxidase biotin (SP) method was used to measure the protein expression of S100A12. The trophoblast cells were cultured in vitro with plasma from the three groups, and a blank control group was set up as well. Transwell was used to detect the cytotrophoblast invasion ability. Western blot was used to measure the protein expression level of receptor for advanced glycation end products (RAGE). Results (1) The levels of S100A12 in maternal peripheral blood of patients with preeclampisa [mild group:(30.8 ± 2.7)μg/L, severe group:(49.3 ± 4.1)μg/L] were significantly higher than that of the control group [(15.8 ± 1.4) μg/L]. In addition, compared with the mild preeclampsia group, the level of S100A12 in the severe preeclampsia group was significantly higher (P<0.05). (2)Positive immunostaining of S100A12 was observed in the cytoplasm of cytotrophoblast, decidual cells and the placentas from the three groups. The positive rate in the mild preeclampisa group was 77%(23/30);in the severe preeclampsia group it was 93%(28/30);and in the healthy pregnant women group it was 23%(7/30). The positive rates of placenta in the mild and severe preeclampsia groups were significantly higher than that in the healthy pregnant women group (P<0.05). In addition, compared with the mild preeclampsia group, the positive rate of immunostaining of S100A12 in the severe group was significantly higher (P<0.05).(3) Cytotrophoblast invasion ability and the expression of RAGE in the mild preeclampsia group were 29.1±3.2 and 0.479 ± 0.038, respectively;in the severe preeclampsia group they were 16.8 ± 2.5 and 0.652 ± 0.059;in the healthy pregnant women group they were 38.6 ± 24.3 and 0.327 ± 0.024; and in the blank control group they were 42.6 ± 5.6 and 0.194 ± 0.011. Cytotrophoblast invasion ability and the expression of RAGE protein in the mild and severe preeclampsia groups were significantly higher than those in the healthy pregnant women group and the control group(P<0.05). Conclusions The expression of S100A12 increased in materal peripheral blood and placenta, and the receptor protein of S100A12 RAGE also had high expression. It suggested that the S100A12 may have some effect on the pathogenesis of preeclampsia.
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Objective To investigate the clinical value of serum soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) in the treatment and therapeutic effect evaluation of patients with an exacerbation of chronic obstructive pulmonary disease.Methods The levels of serum sTREM-1,procalcitonin (PCT) and C-reactive protein (CRP) were determined by enzyme-linked immunosorbent assay (ELISA) in 49 exacerbation of chronic obstructive pulmonary disease (COPD) subjects [acute exacerbation of chronic obstructive pulmonary disease (AECOPD) group],49 stable COPD subjects(sCOPD group) after treatment and 49 healthy volunteers as healthy control group.The levels of sTREM-1,PCT and CRP in different groups were compared and the relationship between the level of sTREM-1 in AECOPD and sCOPD groups,and PCT,and CRP was analyzed,respectively.Results The content of sTREM-1,PCT and CRP between different groups had significant difference(P ＜0.05).The level of sTREM-1 in both AECOPD and sCOPD groups was significantly positive correlated with PCT (P ＜ 0.05) and negative correlated with CRP (P ＞ 0.05).Conclusions For guiding the treatment and curative effect evaluation of patients with AECOPD,sTREM-1 has important clinical reference value.
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Cancer remains the leading cause of death worldwide despite intense efforts in developing innovative treatments. Current approaches in cancer therapy are mainly directed to a selective targeting of cancer cells to avoid potential side effects associated with conventional therapy. In this respect, Natural killer (NK) cells have gained growing attention and are now being considered as promising therapeutic tools for cancer therapy owing to their intrinsic ability to rapidly recognize and kill cancer cells, while sparing normal healthy cells. NK cells play a key role in the first line of defense against transformed and virus-infected cells. NK cells sense their target through a whole array of receptors, both activating and inhibitory. Functional outcome of NK cell against target cells is determined by the balance of signals transmitted from diverse activating and inhibiting receptors. Despite significant progress made in the role of NK cells attack as a pivotal sentinel in tumor surveillance, the molecular has been that regulate NK cell responses remain unclear, which restricts the use of NK cells as a therapeutic measure. Accordingly, current efforts for NK cell-based cancer therapy have largely relied on the strategies that are based on the manipulation of inhibitory receptor function. However, if we better understand the mechanisms governing NK cell activation, including those mediated by diverse activating receptors, this knowledge can be applied to the development of optimal design for cancer immunotherapy by targeting NK cells.
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Activation Analysis , Cause of Death , Immunotherapy , Killer Cells, Natural , Nitriles , Pyrethrins , Receptors, ImmunologicABSTRACT
Objective To investigate difference of NKG2D receptor expression level on the surface of natural killer (NK) cells in the patients with hepatitis B and its clinical significance.Methods This was a four-arm study with different types of subjects,including patients with chronic hepatitis B (CHB,n =22),HBV carriers (HBVC,n=10),patients with acute hepatitis B (AHB,n=18) and healthy donors (HD,n=18).NKG2D protein and mRNA levels on the surface NK cells in the peripheral blood were examined by reverse transcription-polymerase chain reaction assay.The relationship between NKG 2D expression and serum hepatitis B virus (HBV) DNA level was analyzed.The data were compared by analysis of variance and linear regression.Results NKG2D mRNA expression levels in groups of HBVC, HD, AHB and CHB were 0.96±0.17, 1.03±0.12,1.53±0.30 and 1.51 ± 0.35,respectively; the differences among groups were statistically significant (q=7.586,7.485,7.920 and 7.880,respectively; all P＜0.01).NKG2D protein expression levels in groups of AHB,HD,CHB and HBVC were 0.87±0.14,0.89±0.17,0.67±0.09 and 0.59±0.13,respectively; the differences among groups were statistically significant (q=6.92,7.67,7.53and 8.16,respectively; all P＜0.01).The NKG2D mRNA expression levels on NK cells were negatively correlated with serum HBV DNA viral loads in patients with CHB,AHB or HBVC (r=-0.75,-0.66 and-0.69,respectively; all P＜0.01).The NKG2D protein levels on NK cells from patients with AHB and CHB were negatively correlated with serum HBV DNA levels (r=-0.47 and -0.45,respectively; both P＜0.05).Conclusion NKG2D mediated NK cytotoxicity may play a role in viral clearance in hepatitis B.
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Objective To study the roles of advanced glycation end products and its receptor on fetal brain injury of gestational diabetes mellitus (GDM) rats.Methods Twenty one adult pregnant Wistar rats were administered streptozotocin (STZ) intraperitoneally to induce GDM rats model.The fourteen pregnant rats were divided into two groups according to the fasting glucose on the 3rd day of pregnancy:severe GDM group with the fasting glucose ＞ 16.7 mmol/L and mild GDM group with the fasting glucose between 6.7 - 16.7 mmol/L Another seven pregnant rats were chosen as the severe GDM and intervention with micronutrient group,receiving gavage with micronutrient during the whole pregnancy.Five control rats received the same volume of citric acid buffer.All the pregnant rats were tested fasting glucose from the tailvein and their weight on the pregnant day 3,13 and 19.Maternal serum levels of AGE were measured by ELISA and RAGE levels in the embryonic brain tissues were tested by immunohistochemistry.Results ( 1 ) There was no statistically significant difference of pre-pregnancy fasting glucose level among all groups (P ＞ 0.05 ).The fasting glucose levels on the 3rd day and the mean fasting glucouse level of pregnancy in the severe GDM group and the severe GDM and intervention with micronutrient group were higher than those of the control group ( P ＜0.05 ).And there was no significant difference between the severe GDM group and the severe GDM and intervention with micronutrient group (P ＞0.05 ).(2)The serum AGE levels in the severe GDM group and the mild GDM group were( 1037 + 38) ng/L and( 880 ± 34) ng/L respectively,with no significant difference ( P ＞ 0.05 ).The serum AGE levels in the control group and the severe GDM and intervention with micronutrient group were (857 ± 32 ) ng/L and (988 ± 37 ) ng/L,and the difference was statistically significant ( P ＜ 0.05 ).The serum AGE levels in the severe GDM and intervention with micronutrient group and in the mild GDM group had no significant difference ( P ＞ 0.05 ).( 3 ) The serum AGE levels in the severe GDM group,mild GDM group and the control group were positively associated with the mean glucose level of pregnancy ( r =0.603,P ＜ 0.05 ) and the grlucose on the 3rd day of pregnancy (r =0.704,P ＜ 0.05 ).(4)The fetal brain nerve cell number and morphology in the control group were normal.While in the mild GDM group fetal brain nerve cells decreased,the proliferation and swelling of glial cells were seen.In the severe GDM group and the severe GDM and intervention with micronutrient group,the fetal brain cells furtherly reduced,and large vacuole around the cells,deformation and debris of the cells were seen. Glial scar formation was visible in some fetal brain tissues.There was a few RAGE expression in the control fetal brain tissues.In the mild GDM group and the severe GDM group,RAGE expression increased significantly.And the RAGE expression intensity in the severe GDM and intervention with micronutrient group was between the severe and the mild GDM groups.Conclusions( 1 ) Abnormal fetal brain development of GDM rats was associated with the increase of maternal serum AGE and the enhancement of RAGE expression in fetal brain tissues,which suggested that AGE/RAGE pathway may play an important role in the fetal brain injury of GDM rats.(2) Micronutrients can reduce the brain damage of GDM fetuses.
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Objective To investigate the regulatory effect of CsA on the expression of NK cell inhibitory receptor ILT4 and cytotoxicity of NK cells.Methods NK cells treated with CsA ( 10 mg/L) or DMSO for 12,24 and 36 h were chosen as three experimental groups and control groups respectively.RTqPCR and flow cytometry were performed to detect the alteration of ILT4 at the mRNA and protein level respectively.The expression of HLA-G in human gastric cancer cell line BGC-823 and human placental choriocarcinoma cell line JEG-3 were measured at the same time,and then the cytolytic activity of the untreated NK cells and NK cells treated with CsA for 36 h against BGC-823 and JEG-3 cells was determined with MTT.One-way analysis of variance was employed to compare the different ILT4 expression at different time points after medication; Dunnett test was performed to carry out the pairwise comparison between each mean.The difference of HLA-G expression between JEG-3 cells and BGC-823 cells,and the difference of NK cell cytolytic activity against JEG-3 cells and BGC-823 cells were analyzed by student's t-test.Results RT-qPCR assay indicated that the relative levels of ILT4 mRNA in NK cells treated with CsA for 12,24 and 36 h in turn were 0.99 ± 0.27,1.79 ± 0.29,6.79 ± 0.64,and those of their contrast groups treated with DMSO were 0.86 ±0.11,0.94 ±0.12,1.06 ±0.17.The expression of ILT4 in NK cells treated with CsA for 24 h or 36 h was higher than that in NK cells of their contrast groups respectively ( t value of 4.69,14.99,P ＜0.05,respectively),but there was no significant difference between the two groups of NK cells treated for 12 h ( t =0.78,P ＞0.05 ).Through flow cytometry,the positive rates of ILT4 protein expression in NK cells treated with CsA for 12,24 and 36 h [(5.16 ± 0.42 ) ％,( 6.23 ± 0.48 ) ％,( 23.8 ± 1.5 ) ％]were higher than those in NK cells after treatment with DMSO for 12,24 and 36 h respectively[(3.08 ±0.19)％,(3.35 ±0.12)％,(3.36 ±0.21 )％ ;t value of 7.70,10.06,20.72,P＜0.01,respectively].The expression of ILT4 in NK cells treated for 36 h was much higher than that in NK cells for 12 and 24 h at the mRNA and protein level (t value of 16.38,14.12 ;21.81,20.56,P ＜ 0.01,respectively).Meanwhile the killing rates of NK cells treated with 10∶1 effector-target ratio CsA on BGC-823 cells (low HLA-G expression) were ( 8 1.96 ± 2.80 ) ％ ( before treatment) and ( 60.23 ± 1.57 ) ％ ( after treatment),which were higher than those on JEG-3 cells (HLA-G-overexpression) [(53.46 ±2.21 )％ ( before treatment),(28.30 ± 1.85 ) ％ ( after treatment)].The changes of cytotoxicity of NK cells treated with CsA against target cells showed that CsA inhibited the killing activity of NK cells to BGC-823 and JEG-3 cells (t value of 11.74,15.16,P＜0.01,respectively),and the inhibitory rates were (26.48 ±2.42)％ and (47.10 ±1.59 ) ％ respectively.CsA had a higher killing rate inhibition on JEG-3 than on BGC-823 ( t =12.31,P ＜0.01 ).Conclusion CsA induces upregulation of ILT4 in NK cells,and the cytotoxicity of NK cells to tumor cells can be affected by interaction of ILT4 and HLA-G.
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Objective To explore the diagnosis value and the mechanism of triggering receptor expressed on myeloid cells-1 (TREM-1) in early liver damage of severe acute pancreatitis with secondary infection. Methods Twenty-four male SD rats were randomly divided into the control group, the severe acute pancreatitis (SAP) group and the secondary infection of SAP (SISAP) group.The animal model was established by intraperitoneal injection of L- arginine and E. coli. After 24 hours, the serum levels of amylase, glutamate aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor (TNF)-α, C-reactive protein (CRP) and the variation of TREM-1 expression were tested. The blood and peritoneal fluid samples were collected for bacterial culture.Part of the pancreas and liver tissue were taken for histopathological score under microscope. The expression of TREM-1 at mRNA and protein level in liver tissue was detected through Real-time PCR and Western Blot. Results The histological score of pancreas and liver, serum amylase, ALT and AST were significantly higher in the SAP and SISAP groups than those in C group (P<0. 05), and higher in SISAP group than in SAP group (P<0. 05). The CRP and TNF-a expression in SAP and SISAP groups were higher then those in control group, while there was no significant difference between the two groups (P=0. 262 and 0. 359 , respectively). The positive ratio of bacterial culture in blood and peritoneal fluid was 0(0/8), 12. 5% (1/8), and 100% (8/8) in control group, SAP group and SISAP group respectively. The expression of TREM-lmRNA in liver was 2. 10 ± 0. 33 in SAP group and 4. 58+ 1. 00 in SISAP group, which were significantly higher than that in control group (1. 00,P<0. 05) , and the expression of TREM-1 mRNA in SISAP group was higher than that in SAP group (P < 0.05). The expression of TREM-1 at protein level was higher in SISAP group,significantly stronger than that in control and SAP group. Conclusions TREM-1 may play an important role in the early liver damage caused by severe acute pancreatitis.
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Objective To investigate the diagnostic value of soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) in critically ill patients complicated with sepsis. Methods Fifty-six cases of systemic inflammatory response syndrome (SIRS) who admitted in intensive care unit (ICU) of the Second Hospital of Tianjin Medical University between May 2009 and June 2010 were recruited. The SIRS patients were divided into two groups: sepsis group (n= 32) and non-sepsis group (n= 24). The levels of procalcitonin (PCT), C-reactive protein (CRP) were measured within 24 hours of hospitalization.Levels of sTREM-1 were measured by enzyme linked immunoabsorbant assay (ELISA). Based on the receiver operating characteristic curve (ROC), cut-off value, sensitivity and specificity of sTREM-1 in diagnosis of sepsis were calculated. Comparison between groups was done by t test and Mann-Whitney U test. Count data were compared by chi square test. ResultsThe serum level of sTREM-1 were significantly higher in sepsis group than that in non-sepsis group (250.9 ng/L vs 103.6 ng/L, Z= 12. 864,P=0. 002). Areas under the curve (AUC) of sTREM-1, PCT and CRP were 0. 935, 0. 891 and 0. 602, respectively. With a cut-off value of 135.0 ng/L, the sensitivity and specificity of sTREM-1 in diagnosing sepsis were 93. 8％ and 84. 7％, respectively. With a cut-off value of 2. 0 μg/L, the sensitivity and specificity of PCT were 84. 4％and 87. 8％, respectively.ConclusionSerum level of sTREM-1 could be used as an early indicator for diagnosing sepsis.
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Objective To observe the changes of serum soluble form of triggering receptors expressed on myeloid cell-1 (sTREM-1) level in full-term newborns with infection and to investigate the relationship between serum sTREM-1 and neonatal bacterial infection.Methods Eighty-five full-term newborns admitted to the neonatal ward of Shanghai Children s Hospital of Shanghai Jiaotong University were selected into this study.According to the locations and severity of infection,patients were divided into 3 groups: severe infection group (n = 27),mild infection group (n = 28),non-infection group (n = 30).The samples of infection groups were collected before using antibiotics and within 48 h after infection symptom occurred; others were collected during hospitalization.For the neonates with organ dysfunction in the severe infection group,samples were also collected at the third and seventh day of infection.Serum sTREM-1 was measured by enzyme-linked immunosorbent assay.Analysis of variance was used to compare the difference between groups.Receiver operating characteristic (ROC) curve was used to calculate the sensitivity,specificity,positive and negative predictive value and Youden index.Results (1) Serum sTREM-1 level of severe infection group[(91.2±47.3) pg/ml] was significantly higher than that of mild infection group[(68.8 + 30.4) pg/ml] and non-infection group[(35.5±17.6) pg/ml],respectively (P＜0.05).(2) Serum sTREM-1 level of the survival newborns (n= 17) in the severe infection group was lower than that of dead ones[(73.1±34.9) pg/ml vs (121.6±49.3) pg/ml,t= - 2.995,P = 0.006].(3) For the survival patients,the serum sTREM-1 level decreased in the first week of infection,while that of dead patients increased,the cut-off value was 100.6 pg/ml.(4) Based on the ROC analysis,43.8 pg/ml was selected as the the cut-off value,area under the curve was 0.868,and sensitivity was 85.5%,specificity 80.0%,positive predictive value 0.887,negative predictive value 0.750,Youden index 0.655.Conclusions Serum sTREM-1 level increases in neonatal infection.The change of serum sTREM-1 level in patients with severe infection is correlated to the prognosis.
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Objective To investigate the synergistic therapy effects of B and T lymphocyte attenuator(BTLA) extracellular domain in combination with heat shock protein 70 (HSP70)-TC-1 antigen peptide complex on the mouse model of cervical cancer and the related immunological mechanisms. Methods(1)Detecting the BTLA and herpesvirus entry mediator (HVEM) gene expression in the tumor microenvironment after C57BL/6 mice were inoculated with TC-1 tumor cells by realtime PCR; BTLA,HVEM expression on tumor infiltrating lymphocytes cell surface were detected by flow cytometry (fluorescence intensity). (2) According to different treatments, tumor-bearing mice were divided into 5 groups, which was injected with pcDNA3. 1 (empty vector plasmid as control), psBTLA (vector plasmid which expresses BTLA extracellular domain), HSP70 (HSP70-TC-1 cell peptide complex), HSP70 +pcDNA3.1 or HSP70 + psBTLA, respectively. The weight of tumor was recorded. The expression of immunoregulatory genes in tumor microenvironment were detected. The change of lymphocyte amount and cytotoxicity were detected too; lymphocyte proliferation activity was measured by tritium thymidine incorporation assay; the concentration of interleukin (IL) 2 and interferon-γ(IFN-γ) in supernatants of spleen lymphocyte were measured by enzyme-linked immunosorbent assay (ELISA). Results (1) BTLA gene expression was gradually increased after tumor cells inoculation. The highest expression level was 2. 83 + 0. 35 at 14th day, which had statistical significance difference with the 7th day expression of 1.66±0. 25 (P < 0. 05). While HVEM mRNA expression did not change significantly (P > 0. 05). The 7th and 14th day after TC-1 cells inoculation, the average fluorescence intensity of BTLA expression on the surface of tumor infiltrating lymphocytes was 33.5 and 51.8, respectively, in which there was statistically significant difference (P <0. 05); while the difference of HVEM expression was not statistically significant (57. 2 vs 49. 3 ,P >0. 05). (2)The 28th day after inoculation, tumor inhibition rate of HSP70 + psBTLA group was 88%, which was significantly higher than other treatment groups (P <0. 05). The 28th day after TC-1 cells inoculation, combination therapy not only promoted IFN-γ and IL-2 gene (3. 12 + 0.71,3.20 + 0. 62)expression but also reduced transforming growth factor-β (TGF-β), Foxp3 and IL-10 expression (0. 25±0. 03,0. 19 +0. 03,0. 31 +0. 04;P <0. 05). It also promoted CD8+ T lymphocyte infiltration(52 +6)/high power field, cytotoxicity (65.5±2.4) %, proliferation (15.0 × 103 cpm) and cytokine IL-2 , IFN-γsecretion(824±51), (1096±112) pg/ml, which were all significantly higher than other groups (P <0. 05). Conclusion The effect of immunotherapy on tumor can be augmented by the combination of psBTLA which expresses extracellular domain of BTLA and HSP70-TC-1 tumor antigen peptide complex,which could improve the expression of the related immunoregulatory genes to establish a much better microenvironment in favor of anti-tumor immune response against the mice model of the cervix carcinoma.
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Objective To investigate the expression of sialic acid-binding immunoglobulin-like lectin-one (Siglec-1, also called CD169) in lymphocytes, monocytes and neutrophils in peripheral blood in patients with coronary heart disease(CHD), and explore the relationship between Siglec-1 expression and atheresclerosis. Methods CD145 CD169 positive cell proportion and CD169 mRNA levels were respectively measured by flow cytometry and real-time quantitative reverse transcription-polymerase chain reaction (FQ-RT-PCR) in 57 CHD patients and 38 healthy controls. And the levels of serum hpids were determined by automatic biochemistry analyzer. Results The flow cytometry analysis showed that CD169 protein was not found in lymphocytes and neutrophils in both CHD patients and healthy controls. The rate of CD14 CD169 double positive ceils in monocytes in CHD group was significandy higher than that in healthy controls [(12.7±2.4)% vs (1.0±0.3)% ,t =23.2,P<0.01]. And FQ-RT-PCR analysis showed that the mean CD± mRNA copy number in PBMCs in CHD group was significantly higher(3.2 fold) than that in healthy controls [t = 6. 59, P < 0.01]. However, neither differences of CD169 protein positivities [[(12. 2 ± 2. 3) %vs (13.4±2.5)% ,t = 1.87,P >0.05] nor mRNA levels [3.64 fold vs 2.79 fold when compared with healthy controls,t =0. 98, P > 0. 05] were found between CHD patients with normal and abnormal levels of serum Lipids. Conclusions CD169 is mainly expressed in human tissue-resident macrophages but not expressed in peripheral blood monecytes. And when the monocytes is stimulated by inflammation, the expression of CD169 is increased. In patients with CHD, the increased expression of CD169 protein and mRNA level has demonstrated the activation of monocytes in peripheral blood. CD169 and CD169-mediated monocytes activation may play an important role in the development and progression of atherosclerosis.
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Objective To investigate the alterations in killer cell immunoglobulin-like receptors (KIRs)2D and their specific HLA-Cw ligands in patients with ankylosing spondylitis(AS)and determine whether the changes were correlate to the pathogenesis of AS.Methods Polymerase chain reaction of sequence specific primerB(PCR-SSP)was employed for genotyping the presence or absence of five KIR2D genes(KIR2DL1,2DS1,KIR2DL2,2DL3,2DS2)as well as HLA-Cw01-08 alleles from genomic DNA in 105 individuals with AS,together with 51 individuals with osteoarthritis(OA)and 120 healthy controls.Then HLA-C10-08 was divided into two groups.HLA-Cwasn and HLA-Cwlys to calculate the frequency of KIRID genotype.HLA-Gu alleles and KIR/HLA-Cw genotypes.Results The frequencies of HLA-Cwlys genes were significandy higher in patients with AS(0.269 7)compared with those in OA controls(0.148 2)and healthy controls(0.138 8,P=0.024,P=0.001,respectively).The frequency of KIR2DS1/HLA-Cwlys combination Was also markedly higher in AS group(26.67%)than that in OA controls(11.76%)and healthy controls(13.33%,P=0.039,P=0.018,respectively).Condusion The data suggest that the HLA-Cwlys allele may be associated with genetic susceptibility to AS and moreover.in the existence of HLA-Cwlys.the individuals with KIR2DS1 gene are likely to be at increased risk of AS.
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Objective To observe the effects of fenofibrate on the expression of scavenger receptor class B type I(SRBI) in adipose tissue and adipocytes of hypercholesterolemia rabbits.Methods Ten male New Zealand white rabbits were fed with high cholesterol diet for 8 weeks,and were randomly divided into two groups: high cholesterol group and treatment group.The rabbits in the high cholesterol group were maintained cholesterol diet for 4 weeks and those of the treatment group were fed with the same cholesterol diet supplemented with fenofibrate(30 mg/kg/day) for 4 weeks.Five rabbits in control group were fed with normal diet for 12 weeks.Subcutaneous adipose was collected for adipocytes culture.The expressions of SRBI mRNA in adipose tissue and adipocytes were evaluated by semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR).Results The serum levels of total cholesterol and low density lipoprotein cholesterol were significantly higher in the high cholesterol group and treatment group than those of the control group(P0.05).The expressions of SRBI mRNA in adipose tissue and adipocytes were up-regulated in high cholesterol group compared with those of control group(P