Association between the level of CD4^+T lymphocyte microRNA-155 and coronary artery disease in patients with unstable angina pectoris

Objective To study the association between the expression of microRNA-155(miRNA-155)in peripheral blood CD4^+T lymphocytes and the level of semrn interferon-7(IFN-7)concentration and the severity of coronary artery disease (CAD).Methods After coronary angiography,252patients with suspected unstable angina pectoris (UAP)were divided into the UAP group (128patients with CAD confirmed by angiography)and the control group (124patients without CAD confirmed by angiography).Fresh peripheral blood was extracted 16-24h before coronary angiography,CD4^+T lymphocytes was tested using immunomagnetic beads,the expression ofmiRNA-155was tested using quantitative PCR and the expression of IFN-7was tested using enzyme-linked immunosorbent assay (ELISA).According to the results of angiography,Gensini score of coronary artery lesions was analyzed.Furthermore,we also analysis the association between the level of miRNA-155in peripheral blood CD4^+T lymphocytes,the level of serum IFN-γand Gensini score of coronary lesion.Results The levels ofmiRNA-155(0.49±0.08vs.0.23±0.09)and IFN-7(227.58±26.01vs.141.23±17.89)in the UAP group were significantly higher than that of the control group,the difference was statistically significant.The level of miRNA-155and IFN-γwere positively correlated with Gensini score of CAD (r =0.534,r =0.713,respectively,all P <0.05).The level of miRNA-155was positively correlated with the level of IFN-γ,(r =0.686,P <0.05).Conclusions The level of miRNA-155in peripheral blood CD4^+T lymphocytes and the level of IFN-γ are closely correlated with the severity of CAD.

黑色素瘤B16细胞通过释放外泌体诱导naïve CD4^(+) T细胞分化为Treg

目的检测小鼠黑色素瘤细胞B16通过释放外泌体诱导naïve CD4^(+)T细胞分化为调节性T细胞(regulatory T cell,Treg)的能力。方法通过试剂盒和超速离心获得B16培养上清中的外泌体,用透射电镜观察外泌体形态,Western blot检测外泌体标记性蛋白的表达水平,通过Transwell共培养验证外泌体或B16细胞对naïve CD4^(+)T细胞诱导分化的能力,流式细胞术检测小鼠脾脏、肿瘤内Th17、Treg细胞的频率,qPCR检测T细胞亚群特异性转录因子、炎症因子的mRNA水平。结果B16细胞外泌体的直径为30~100 nm,呈圆形碟状结构,外泌体高表达Tsg101、HSP60和Alix,低表达GAPDH;在体外,B16细胞外泌体可直接诱导naïve CD4^(+)T细胞分化成Treg,但不能诱导其分化为Th17;小鼠在体实验证实,B16细胞外泌体能够诱导Treg细胞并聚集于小鼠脾脏中,同时能够增加B16瘤体内Treg细胞的数量,并活化使其表达更高水平的IL-10、TGF-β和Foxp3。结论B16细胞外泌体通过诱导naïve CD4^(+)T细胞分化为Treg,并将其募集至肿瘤组织,从而维持肿瘤的免疫抑制微环境。

Changes of CD4^+CD25^+ Regulatory T Cells in Patients with Acute Coronary Syndrome and the Effects of Atorvastatin

The function of CD4＋CD25＋ regulatory T lymphocytes （Treg） in patients with acute coronary syndrome （ACS） and the effects of atorvastatin were investigated. Forty-eight patients with ACS were randomly divided into two groups： group C receiving conventional therapy （n=24）, and group C＋A receiving conventional therapy＋atorvastatin （10 mg/day, n=24）. T lymphocytes from ACS patients （before and 2 weeks after the treatment） or 18 healthy subjects were separated and the flow cytometry was used to measure the percentage of Treg. The inhibitory ability of Treg on effector T cells was determined by mixed lymphocyte reaction （MLR）. ELISA was used to measure the serum levels of cytokines （IL-10, TGF-β1 and IFN-γ） before and after treatment. The results showed that as compared with normal control group, Treg percentage was decreased significantly （P〈0.01）, the inhibitory ability of Treg on the T lymphocytes proliferation was reduced （P〈0.01）, IFN-γ levels were increased and IL-10 and TGF-β1 levels were lowered in ACS patients. After treatment with atorvastatin, Treg percentage and the inhibitory ability of Treg on T lymphocytes proliferation were significantly increased in ACS patients. Serum IFN-γ was decreased significantly, while IL-10 and TGF-β1 were elevated significantly as compared with the non-atorvastatin group. The number of Treg was positively correlated with serum TGF-β1, but negatively with serum IFN-γ and CRP. It was concluded that ACS was associated with decreased number and defected function of Treg, which may play an important role in initiating immune-inflammatory response in ACS. The inhibitory effects of atorvastatin on inflammation in ACS may be due to its beneficial effects on Treg and restoration of immune homeostasis.

Effects of pituitary adenylate cyclase activating polypeptide on CD4^+/CD8^+T cell levels after traumatic brain injury in a rat model

BACKGROUND： The effect of pituitary adenylate cyclase activating polypeptide （PACAP） during traumatic brain injury （TBI） and whether it can modulate secondary injury has not been reported previously. The present study evaluated the potential protective effects of ventricular infusion of PACAP in a rat model of TBI.METHODS： Male Sprague Dawley rats were randomly divided into 3 treatment groups （n=6, each）： sham-operated, vehicle （normal saline）＋TBI, and PACAP＋TBI. Normal saline or PACAP （1 μg/5 μL） was administered intracerebroventricularly 20 minutes before TBI. Right parietal cortical contusion was produced via a weight-dropping method. Brains were extracted 24 hours after trauma. Histological changes in brains were examined by HE staining. The numbers of CD4＋ and CD8＋ T cells in blood and the spleen were detected via flow cytometry.RESULTS： In injured brain regions, edema, hemorrhage, inflammatory cell infiltration, and swollen and degenerated neurons were observed under a light microscope, and the neurons were disorderly arrayed in the hippocampi. Compared to the sham group, average CD4＋ CD8＋ lymphocyte counts in blood and the spleen were significantly decreased in rats that received TBl＋vehicle, and CD4- CD8＋ were increased. In rats administered PACAP prior to TBI, damage was attenuated as evidenced by significantly increased CD4＋, and decreased CD8＋, T lymphocytes in blood and the spleen.CONCLUSION： Pretreatment with PACAP may protect against TBI by influencing periphery T cellular immune function.

Increase of CD4^+CD25^+ T cells in Smad3^(-/-) mice

AIM： To investigate the changes of lymphocyte subpopulations, especially CD4^＋CD25^ T regulatory cells in Smad3^-/- mice. METHODS： Hematological changes and changes of lymphocyte subpopulations were detected in Smad3＂/- mice using cell counter and flow cytometry, respectively, and compared to their littermate controls. RESULTS： The numbers of neutrophils and lymphocytes in peripheral blood were significantly increased in Smad3^-/- mice compared to littermate controls. CD19^＋ expressing cells in blood and spleen, and CD8^＋ T cells in thymus were all markedly decreased in Smad3^-/- mice. More important, Smad3^-/- mice had an increased population of CD4^＋CD25^＋ T cells in peripheral lymphoid tissues, including thymus, spleen, and lymph nodes. CONCLUSION： These observations suggest that the changes of lymphocyte subpopulations might play a role in susceptibility to inflammation of Smad3^-/- mice.

Haematology Parameters of Apparently Healthy Prospective Whole Blood Donors in a Nigerian Hospital Setting

Background: Adequate selection of a prospective whole blood donor protects his health and safety of the recipient. Objectives: The main objective of this study was to determine the haematology parameters of apparently healthy prospective whole blood donors. Participants and Methods: This was a hospital based prospective study carried out from August to October 2020 at the blood transfusion unit of the Lagos State University Teaching Hospital (LASUTH), Ikeja, Nigeria. A structured pretested questionnaire was used for data collection. The socio demographic status and the haematology parameters of apparently healthy prospective whole blood donors who tested negative for HIV, hepatitis B and C markers were captured. Obtained data were analysed with the statistical package for the social scientist software version 20. Results: One hundred male (97.1%) and three female (2.9%) apparently healthy prospective whole blood donors were studied. The median age of study subjects was 30 years. Obtained median haematology parameter values were 13 g/dl, 40%, 4.9/nl and 203.9/nl for haemoglobin concentration, haematocrit, total white cell and platelet counts respectively. The median values for the mean corpuscular haemoglobin concentration (MCHC), mean corpuscular haemoglobin (MCH) and mean corpuscular volume (MCV) of participants were 32.6 g/dl, 27.7 pg and 85.7 fl respectively. Observed prevalence of subnormal haematology parameters for haemoglobin concentration, total white cells, platelets were 12.6%, 25.2%, and 13.6% respectively. Also subnormal values for MCHC, MCH, MCV were 11.7%, 26.2%, and 16.5% respectively among prospective whole blood donors in this study. No higher than normal haematology parameter values were observed. Median values for erythrocyte sedimentation rate was 8.4 mm/hr. Conclusion: A significant percentage of apparently healthy prospective whole blood donors had subnormal haematology parameters values. Obtained normal values in our study are comparable with local reference range reports from previous studies in Nigeria and other parts of Africa. 124947 .

INVESTIGATION OF REGULATORY T CELLS IN PATIENTS WITH NON-SMALL CELL LUNG CANCER

Objective To evaluate the prevalence of CD4 ^＋ CD25^high regulatory T cells （ Treg cells） in the peripheral blood mononuclear cells （PBMC） and tumor-infiltrating lymphocytes （TIL） of patients with non-small cell lung cancer （NSCLC） and to investigate immunosuppression to the progression of cancer. Methods Peripheral blood and tumor tissues were collected from 20 patients with NSCLC at the time of surgery. None of the patients received surgery, radiotherapy, chemotherapy, or other medical interventions before this study. Cancer stages of the patients were Ⅰ-Ⅲ A. Venous blood samples were obtained from 20 health donors. PBMC were isolated from blood samples by differential centrifugation over Ficoll-Hypaque. TILs were isolated from tumors by differential centrifugation over Ficoll-Hypaque and Percoll. Percentage of CD4^＋ CD25^highTr/CD4＋T in PBMC and TIL was assessed by the flow cytometry. Results The percentage of CD4^ ＋ CD25high Tr/ CD4 ^＋T in PBMC [ （4. 87 ± 1.22 ） % ] of NSCLC patients was significantly higher than that in healthy donors [ （ 2.36 ± 0. 72 ） % ] （ P 〈 0.01 ）. The percentage of CD4^＋ CD25^highTr/ CD4^＋ T in PBMC [ （5.40 ± 1.20） % ] of NSCLC patients in stage Ⅱ-Ⅲ A was significantly higher than that in stage Ⅰ [ （3. 87 ± 0. 22 ） % ] （ P 〈 0. 01 ）. The percentage of CD4 ＋ CD25hiShTr/ CD4 ＋ T in TIL[ （ 8. 66 ±0. 76） % ] of NSCLC patients in stage Ⅱ-Ⅲ A was significantly higher than that in stage Ⅰ [ （ 7. 04 ± 0. 80） % ] （ P 〈 0. 01 ）. Conclusion The prevalence of CD4 ^＋ CD25^highTreg cells in PBMC and TIL of NSCLC patients was significantly higher than that in healthy donors. These Treg cells may be preventing appropriate antitumor immune responses. The population of CD4^ ＋ CD25^highTreg cells in PBMC and TILs of NSCLC patients with Ⅱ-Ⅲ A stage was significantly higher than that of NSCLC patients with Ⅰ stage. These Treg cells may facilitate development of tumors.

CD70 defines a subset of proinflammatory and CNSpathogenic TH1/TH17 lymphocytes and is overexpressed in multiple sclerosis

activation.Therefore,engagement of the costimulatory CD27/CD70 pathway is solely dependent on upregulation of CD70.However,the T cell-intrinsic effect and function of human CD70 remain underexplored.Herein,we describe that CD70 expression distinguishes proinflammatory CD4^(+)T lymphocytes that display an increased potential to migrate into the central nervous system(CNS).Upregulation of CD70 on CD4^(+)T lymphocytes is induced by TGF-β1 and TGF-β3,which promote a pathogenic phenotype.In addition,CD70 is associated with a TH1 and TH17 profile of lymphocytes and is important for T-bet and IFN-γexpression by both T helper subtypes.Moreover,adoptive transfer of CD70−/−CD4^(+)T lymphocytes induced less severe experimental autoimmune encephalomyelitis(EAE)disease than transfer of WT CD4^(+)T lymphocytes.CD70+CD4^(+)T lymphocytes are found in the CNS during acute autoimmune inflammation in humans and mice,highlighting CD70 as both an immune marker and an important costimulator of highly pathogenic proinflammatory TH1/TH17 lymphocytes infiltrating the CNS.

CTL Response Suppression in Chronic Phase of Infection:A mathematical study

Human immunodeficiency virus(HIV)has had an insightful impact about the state of healthiness of human immune system.Due to great improvement in drug therapy,HIV infections have been reduced by 17%over the past eight years.It has been proved that most effective treatment HAART(Highly Active Anti Retroviral Therapy)mainly controls the diseases progression but it does not eradicate the diseases completely.Reverse Transcriptase Inhibitor drugs specially associated with virus specific Cytotoxic T-Lymphocyte(CTL)that declines with disease progression. CTL responses against AIDS pathogenesis could be potential in the dynamics of virus replication, recognition and clearance of infected cells.In this research article a mathematical model has been proposed on the basis of CTL response suppression in the chronic phase of infection due to presence of virus.We also consider the growth of the virus population from the infected CD4^+T cells budding process and from the other infected cells like macrophages and thymocytes.Our analytical and numerical studies are consistent with existing observations from allied areas.

Changes in peripheral blood inflammatory factors(TNF-α and IL-6) and intestinal flora in AIDS and HIV-positive individuals

Objective:In this study,we investigated the changes in peripheral blood inflammatory factors and intestinal flora in acquired immune deficiency syndrome(AIDS)and human immunodeficiency virus(HIV)-positive individuals(AIDS/HIV patients),and explored the relationships among intestinal flora,peripheral blood inflammatory factors,and CD4^+T lymphocytes.Methods:Thirty blood and stool samples from an AIDS group and a control group were collected.The levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were determined by enzyme-linked immunosorbent assay(ELISA),and the number of CD4^+T lymphocytes by a FACSCount automated instrument.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to determine the messenger RNA(mRNA)levels of Bifidobacterium,Lactobacillus,Escherichia coli,Enterococcus faecalis,and Enterococcus faecium.Correlations among intestinal flora,inflammatory factor levels,and CD4^+T lymphocyte values were evaluated using the Spearman correlation coefficient.Results:The levels of TNF-αand IL-6 in the AIDS group were higher than those in the control group,while the number of CD4^+T lymphocytes was lower.The amounts of Bifidobacterium and Lactobacillus in the AIDS group were significantly lower than those in control group,while the amounts of E.coli,E.faecalis,and E.faecium were much higher.The amounts of Bifidobacterium and Lactobacillus were negatively correlated with the content of TNF-αand IL-6 and the CD4^+T lymphocyte count,while those correlations were reversed for E.coli,E.faecalis,and E.faecium.Conclusions:The intestinal microbiota of AIDS/HIV patients were disordered,and there was a correlation between the amount of intestinal flora and the number of CD4^+T lymphocytes and the levels of TNF-αand IL-6.

Clinical laboratory features of Meigs’syndrome:a retrospective study from 2009 to 2018

Meigs’syndrome(MS),a rare complication of benign ovarian tumors,is easily misdiagnosed as ovarian cancer(OC).We retrospectively reviewed the clinical laboratory data of patients diagnosed with MS from 2009 to 2018.Serum carbohydrate antigen 125 and HE4 levels were higher in the MS group than in the ovarian thecoma-fibroma(OTF)and healthy control groups(all P<0.05).However,the serum HE4 levels were lower in the MS group than in the OC group(P<0.001).A routine blood test showed that the absolute counts and percentages of lymphocytes were significantly lower in the MS group than in the OTF and control groups(all P<0.05).However,these variables were higher in the MS group than in the OC group(both P<0.05).The neutrophil-to-lymphocyte ratio(NLR)was also significantly lower,whereas the lymphocyte-to-monocyte ratio was higher in the MS group than in the OC group(both P<0.05).The NLR,platelet-to-lymphocyte ratio,and systemic immune index were significantly higher in the MS group than in the OTF and control groups(all P<0.05).The hypoxia-inducible factor-1 mRNA levels were also significantly higher,whereas the glucose transporter 1,lactate dehydrogenase,and enolase 1 mRNA levels were lower in peripheral CD4+T cells obtained preoperatively in a patient with MS than those in patients with OTF,patients with OC,and controls(all P<0.05).The expression of these four glucose metabolism genes was preferentially restored to normal levels after the tumor resection of MS(P<0.001).These clinical laboratory features can be useful in improving the preoperative diagnostic accuracy of MS.