ABSTRACT
@#Objective To analyze the topology of IFN-induced transmembrane(IFITM) protein in porcine peripheral blood lymphocytes(PBMCs) and detect the change of IFITM mRNA transcription in PBMCs after porcine reproductive and respiratory syndrome virus(PRRSV) infection in vitro.Methods PRRSV,porcine circovirus 2(PCV2) and Japanese encephalitis virus(JEV) negative anticoagulant blood of piglets were collected aseptically and isolated for PBMCs.Porcine IFITM CDS sequence was amplified by PCR,sequenced and analyzed for topology.PBMCs were infected with PRRSV in vitro.Cell samples were collected at 12,24,36 and 48 h after infection,detected for PRRSV infection by RT-PCR,and detected for mRNA transcription level changes of IFITM1,IFITM2 and IFITM3 by RT-PCR.Results The porcine PBMCs were successfully isolated and the full-length sequence of IFITM CDS derived from PBMCs was cloned.The porcine IFITM protein might have two topological structures.PBMCs inoculated with PRRSV for 24 h produced obvious cytopathic effect.PRRSV was replicated in PBMCs.The transcription levels of IFITM1,IFITM2 and IFITM3 mRNA in PBMCs were significantly up-regulated at the early stage of PRRSV infection,and reached the peak at 12h after infection,and then gradually decreased;The transcription level of IFITM1 mRNA increased at 36 h after virus infection and then declined rapidly.Conclusion PRRSV infection in vitro significantly up-regulated the transcription level of IFITM mRNA in PBMCs,indicating that IFITM was involved in the antiviral immune response of PBMCs.This study provided a reference for revealing the natural immune response against PRRSV in vivo.
ABSTRACT
Objective:To investigate chromosomal aberrations in peripheral blood lymphocytes of underground miners, in order to explore the influencing factors involved in chromosomal aberration levels of non-uranium metal mines.Methods:Totall 135 workers were recruited from an iron mine and a gold mine located in different cities of Henan province, where 69 workers worked aboveground and 66 miners worked underground in the metal mines. The radon concentration in the mines was measured by solid-state nuclear track detectors. Chromosomal aberrations in peripheral blood lymphocytes from the subjects were detected using conventional analysis method, and the influence factors of chromosomal aberrations were analyzed.Results:Radon concentration was 30-2 943 Bq/m 3 in the aboveground workplace of the mines, and 62-28 314 Bq/m 3 in underground. The age of the underground group was obviously lower than that of the aboveground group( t=2.12, P<0.05), but the frequencies of dicentrics, translocation, acentric fragment, and total chromosome-type aberrations in the underground group were significantly higher than those in the aboveground group ( χ2=10.49, 16.74, 8.15, 29.50, P<0.01). Consistent results were obtained when only male workers were regarded as object of observation ( χ2=8.44, 11.63, 4.94, 20.81, P<0.05). The frequency of translocation ( χ2=8.44, P<0.05) was dependent on the length of service in the underground group. Poisson regression analysis indicated that the aboveground and undergroud grouping partly affected the levels of dicentrics, translocation, acentric fragment, and total chromosome-type aberrations (the underground group IRR=3.25, 2.69, 1.97, 2.18, P<0.05). Conclusions:The radon exposure in the underground workplace of the metal mines may be the main factor resulting in the increase of chromosome-type aberrations of miners. The occupational health and safety of the miners who may be exposed to high radon levels are worthy of great attention.
ABSTRACT
【Objective】 To investigate the prevalence of occult hepatitis C virus infection (OCI) in Shaanxi blood donors and characterize the occult hepatitis C virus in this cohort. 【Methods】 Between July and September of 2019, 112, 117, 46, and 75 blood samples were obtained from eligible donors, alanine aminotransferase (ALT) elevated donors(ALT>50 U/L), hepatitis B virus surface antigen (HBsAg) positive donors, and anti-hepatitis C virus (anti-HCV) positive donors, respectively. Peripheral blood lymphocytes (PBMCs) were isolated from all the samples. HCV 5′UTR was amplified from total RNA of PBMCs using reverse transcription nested PCR (RT-Nested PCR) to detect the infection status of OCI in blood donors. HCV RNA core/E2 region was amplified from HCV RNA positive samples with RT-Nested PCR to determine HCV genotypes and subtypes via sequencing. 【Results】 Two (2.67%, 2/75) cases of OCI were identified in anti-HCV positive samples, 1(0.85%, 1/117) case of OCI was identified in abnormal ALT samples, and all OCI were genotyped as 1b. OCI was not found in HBsAg positive and eligible blood donors. 【Conclusions】 Our findings suggest that OCI does exist among Shaanxi blood donors and that ALT may be useful in diagnosing OCI.
ABSTRACT
OBJECTIVE: To explore the effect of Radix Ranuncoli Ternati on the expression of granule lytic peptide and bactericidal ability of T lymphocyte in peripheral blood lymphocytes of patients with pulmonary tuberculosis. METHODS: Peripheral blood lymphocytes were treated with Ranunculus. MTT assay was used to detect cell proliferation. Real-time PCR was used to detect the mRNA level of GLS in patients. RESULTS: Radix Ranuncoli Ternati could enhance expression of Tubercle bacillus. The RESULTS of MTT assay indicated that the rate of peripheral blood lymphocytes apoptosis inducted by the Radix Ranuncoli Ternat. The Western blotting RESULTS showed that Radix Ranuncoli Ternati made the expression levels of GLS in peripheral blood lymphocyte increasing. When the concentration of the drug is 200 mg·L-1, it can effectively induce the activation of tuberculosis resting bacteria. The expression of GLS and the level of mRNA were increased in peripheral blood lymphocytes. CONCLUSION: Radix Ranuncoli Ternati may increase the expression of GLS and enhance the body's killing effect on pathogenic bacteria to achieve the therapeutic effect.
ABSTRACT
ABSTRACTINTRODUCTION:While no single factor is sufficient to guarantee the success of influenza vaccine programs, knowledge of the levels of immunity in local populations is critical. Here, we analyzed influenza immunity in a population from Southern Brazil, a region with weather conditions that are distinct from those in the rest of country, where influenza infections are endemic, and where greater than 50% of the population is vaccinated annually.METHODS:Peripheral blood mononuclear cells were isolated from 40 individuals. Of these, 20 had received the H1N1 vaccine, while the remaining 20 were unvaccinated against the disease. Cells were stimulated in vitro with the trivalent post-pandemic influenza vaccine or with conserved major histocompatibility complex I (MHC I) peptides derived from hemagglutinin and neuraminidase. Cell viability was then analyzed by [3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide)]-based colorimetric assay (MTT), and culture supernatants were assayed for helper T type 1 (Th1) and Th2-specific cytokine levels.RESULTS:Peripheral blood lymphocytes from vaccinated, but not unvaccinated, individuals exhibited significant proliferation in vitro in the presence of a cognate influenza antigen. After culturing with vaccine antigens, cells from vaccinated individuals produced similar levels of interleukin (IL)-10 and interferon (IFN)-γ, while those from unvaccinated individuals produced higher levels of IFN-γ than of IL-10.CONCLUSIONS:Our data indicate that peripheral blood lymphocytes from vaccinated individuals are stimulated upon encountering a cognate antigen, but did not support the hypothesis that cross-reactive responses related to previous infections can ameliorate the immune response. Moreover, monitoring IL-10 production in vaccinated individuals could comprise a valuable tool for predicting disease evolution.
Subject(s)
Adult , Humans , Young Adult , Antibodies, Viral/immunology , Influenza A Virus, H1N1 Subtype/immunology , Influenza Vaccines/immunology , Influenza, Human/immunology , Lymphocytes/immunology , Antibodies, Viral/blood , Brazil/epidemiology , /immunology , Cross-Sectional Studies , Influenza, Human/epidemiology , Influenza, Human/prevention & control , Interferon-gamma/biosynthesis , /biosynthesis , Leukocytes, Mononuclear/immunology , PandemicsABSTRACT
Patients treated with radiotherapy (RT) might experience a large variation in normal tissues. Severe radiation damage in a minority of patients limits the doses that might be safe given to the majority. The possibility of predicting such radiation-induced damage would provide a better treatment schedule for the patients. Several predictive tests in peripheral blood lymphocytes such as initial DNA damage, radiation-induced apoptosis and genetic variation have been proposed to know the individual sensitivity of patients to the radiotherapy schedules. This study aimed to summarize the main studies regarding to this ifeld.
ABSTRACT
Objective To detect the expression of DNA damage response genes induced by radiation in human peripheral blood lymphocyte,and to explore the new biomarkers of radiation.Methods The human peripheral blood cells were irradiated to X-rays at different doses of 0,1,2,3,4,and 5 Gy.The quantitative real.time qPCR wag used to detect the expressions of cyclin-dependent kinase inhibitor l a gene(Cdknl a)and growth arrest and DNA damage inducible gene(Gadd45a)in lymphoeytes at 4 and 24 h post-irradiation,respectively.The method of CB mieronucleus was used to determine the change of micronucleus ratio.Results The expression of Cdknl a in peripheral blood lymphocytes wag increased significantly at 4 and 24 h post-irradiation to 0-5 Gy.reached the peak at 4 Gy and began to decrease at 5 Gy,which showed a dose-dependent manner(r=0.946,0.975,P<0.05).Similarly,the expression of Gadd45α in human peripheral blood lymphocytes was also increased significantly at 4 and 24 h post-irradiation to 0-5 Gy in a dose-dependent manner,while the expression of Gadd45a at 4 h wag higher than that at 24 h(r=0.936,0.797,P<0.05).The ratio of micronuclei wag increased significantly at 4 and 24 h post-irradiation to 0-5 Gy(r=0.990,0.984,P<0.05).Conciusions Cdknl a and Gadd45α expression could be increaged significandy at 4 and 24 h post-irradiation to 0-5 Gy,showing a good linear relationship.which might be candidate for radiation biological dosimeter.
ABSTRACT
DNA damage response (DDR) in different cell cycle starus of human peripheral blood lymphocytes (PBLs) and the role of H2AX in DDR were investigated.The PBLs were stimulated into cell cycle with phytohemagglutinin (PHA).The apoptotic ratio and the phosphorylation H2AX (S139)were flow cytometrically measured in resting and proliferating PBLs after treatment with camptothecin (CPT) or X-ray.The expressions of γH2AX,Bcl-2,caspase-3 and caspase-9 were detected by Western blotting.DDR in 293T cells was detected after H2AX was silenced by RNAi method.Our results showed that DNA double strand breaks (DSBs) were both induced in quiescent and proliferating PBLs after CPT or X-ray treatment.The phosphorylation of H2AX and apoptosis were more sensitive in proliferating PBLs compared with quiescent lymphocytes (P＜0.05).The expression levels of anti-apoptotic proteins Bcl-2 were reduced and cleaved caspase-3 and caspase-9 were increased.No significant changes were observed in CPT-induced apoptosis in 293T cells between H2AX knocking down group and controls.It was concluded that proliferating PBLs were more vulnerable to DNA damage compared to non-stimulated lymphocytes and had higher apoptosis rates.γH2AX may only serve as a marker of DNA damage but exert no effect on apoptosis regulation.
ABSTRACT
BACKGROUND: The impact of microwave (MW)/radio frequency radiation (RFR) on important biological parameters is probably more than a simply thermal one. Exposure to radio frequency (RF) signals generated by the use of cellular telephones have increased dramatically and reported to affect physiological, neurological, cognitive and behavioural changes and to induce, initiate and promote carcinogenesis. Genotoxicity of RFR has also been reported in various test systems after in vitro and/or in vivo exposure but none in mobile phone users. AIMS: In the present study, DNA and chromosomal damage investigations were carried out on the peripheral blood lymphocytes of individuals using mobile phones, being exposed to MW frequency ranging from 800 to 2000 MHz. METHODS: DNA damage was assessed using the single cell gel electrophoresis assay and aneugenic and clastogenic damage by the in vivo capillary blood micronucleus test (MNT) in a total of 24 mobile phone users. RESULTS: Mean comet tail length (26.76 ± 0.054 mm; 39.75% of cells damaged) in mobile phone users was highly significant from that in the control group. The in vivo capillary blood MNT also revealed highly significant (0.25) frequency of micronucleated (MNd) cells. CONCLUSIONS: These results highlight a correlation between mobile phone use (exposure to RFR) and genetic damage and require interim public health actions in the wake of widespread use of mobile telephony.
ABSTRACT
Objective To explore the apoptosis and proliferation of peripheral blood lymphocytes(PBLs) from primary nephrotic syndrome(PNS) and effects of dexamethasone(Dex) on them.Methods Minimal change NS(MCNS), non-minimal change NS(NMCNS) and healthy children were involved in this study. PBLs were cultured in vitro with Dex or PHA+Dex or without PHA and Dex. Apoptosis of PBLs was measured by propidium iodide(PI) staining; The effects of Dex at different concentrons on PBLs′proliferation were investigated by 3H-TdR incorporation.Results The apoptotic rate of vacuity group in MCNS was lower compared with NMCNS and healthy controls(P
ABSTRACT
There are lot of studies on the cytogenetic changes in solid tumours but there are very few studies on the cytogenetic changes in peripheral blood by mphosytes in patients with malignancy. In the present study we evaluated peripheral blood lymphocyte cytogenetic changes on short term culture. Twenty four out of 250 patients with various malignancies showed some aberration of karyotypes in peripheral lymphocytes (9-6%) suggesting an underlying genetic instability in these cancer patients or alternatively demonstrating that these changes could be related to exposure to environmental mutagens.
ABSTRACT
Objective To explore the correlation between the expression of lung resistance-related protein (LRP) in non-small cell lung cancer (NSCLC) and that in peripheral blood lymphocytes (PBL). Methods The S-P immunohistochemistry was used to detect the expression of LRP in specimens and PBL of 49 patients with NSCLC. Para-carcinoma tissues and PBL of 10 normal people were used as controls. Results The positive expression rate of LRP was 78.2%, 46.4% and 66.8% in carcinoma tissues, para-carcinoma tissues and PBL, respectively. The expression rate of LRP in carcinoma tissues was higher than that in para-carcinoma tissues (P
ABSTRACT
An aphidicolin is a chemical agent which selectively inhibits DNA polymerase alpha in S phase of cell cycle. The purpose of this study is toinvestigate of chromosomal abnormalities including fragile sites induced by 0.2 microgram/ml and 0.4 ng/ml aphidicolin in lymphocyte cultures of six healthy individuals. The results were follows. 1. A significant decreasing in mitotic indexes in respect to control culture was observed with both aphidicolin concentrations used. 2. The cells showing chromosome aberrations and the total number of cytogeneticic alterations were significantly increased both aphidicolin treated cultures than control cultures. 3. The total numbers of chromosomal aberrations were increased in the concentration of 0.4 microgram/ml aphidicolin compared to 0.2 microgram/ml treated groups. 4. The most frequent type of chromosomal aberration is a gap. 5. A site showing a gap or break was defined as common fragile sites (c-fra) if it appeared more than 1% of cells analyzed and in at least three of six individuals studied with the same culture treatment. Using these criteria, 3p14, 4q12, 5p13, 6q16, 9p13, and 16q23 were induced in different proportions by different concentration of aphidicolin and four of these c-fras, 4q12, 5p13, 6q16, 9p13 have not been reported so far. This results support that aphidicolin induced fragile sites differently according to cultured cell or cultured conditions, and also suggest the mechanism that common fragile sites caused be closely related with the defect of DNA synthesis in the S phase of cell cycle.
Subject(s)
Humans , Aphidicolin , Cell Cycle , Cells, Cultured , Chromosome Aberrations , Cytogenetics , DNA , DNA Polymerase I , Lymphocytes , Mitotic Index , S PhaseABSTRACT
To investigate fragile sites induced by aphidicolin which is a specific inhibitor of eukaryotic DNA polymerase a which is primarily associated with chromosomal DNA replication in human lymphocytes, HaCat cells (human keratinocytes) and MRC-5 cells (human embryonic lung fibroblast), we cultured each cells in RPMI 1640 with 10% fetal calf serum and 2% PHA. Treatment of the cells with aphidicolin was generally carried out for the last 24 hours of culturing. The drug was dissolved in DMSO and used at final concentrations of 0.05~0.15 mg/ml, corresponding to a maximum DMSO concentration of 0.028%. Karyotypes of each cells were performed by routine method, and 50 metaphases were scored for each culture for analysis of breakage rate. Experimental cells treated with APC showed a dose dependent sensitivity and the amounts of chromosome breakage induced by APC are the highest in concentration of 0.15 mg/ml. The frequency of fragile sites on each cells appeared in MRC-5 cells, lymphocytes and HaCat cells in order. The common fragile sites on all experiments was 16q23, and the common fragile sites on embryonic cells was 1p31. It can be concluded that gene or nucleic acid which is located on 16q23 is the most important factor to induce chromosomal breakage with sensitivity to aphidicolin and 1p31 is important site to induce chromosomal breakage in embryonal cells.
Subject(s)
Humans , Aphidicolin , Chromosome Breakage , Dimethyl Sulfoxide , DNA , DNA Replication , Karyotype , Lung , Lymphocytes , MetaphaseABSTRACT
The gene expressions of Interleukin-6 (IL-6) from human peripheral blood lymphocytes (HPBL) stimulated by C. albicans were investigated by using ELISA (Enzyme linked immunosorbent assay), reverse transcription polymerase chain reaction (RT-PCR) and northern blotting. HPBL (1 X 10'/ml) obtained from normal human peripheral blood lymphocytes were cultured with live C. albicans (LCA) or heat killed C. albicans type A 311 (KCA, 3 X 10/ml) for various times (0.5, 1, 4, 8, 18, 24, 48 and 72 hours). On the purpose of this experiment, we also used lipopolysacchalide (LPS, 10 ug/ml), zymosan (1, 10, 100 ug/ml) as a polysacchaide component of the wall of yeast cells or TNFa (50, 100 ng/ml) as a IL-6 inducers. For observation of the level of IL-6 gene expression, actinomycin D (AD, 5 pg/ml) or cyclohexamide (CHX, 25 ug/ml) was added to HPBL stimulated with LCA for 0.5, 2, 4 hours and the HPBL were assessed for IL-6 mRNA. The highest value for IL-6 activity by LCA were observed at 48 hours reaction, but in the case of KCA, highest value of IL-6 activity was observed at 72 hours reaction and the value was also higher (500 pg/ml) than that of LCA (188 pg/ml). 1L-6 mRNA induced by LCA were detected up to 48 hours but in the case of KCA, the band for IL-6 mRNA were far stronger and appeared until lately than that of LCA. Therefore, the results of IL-6 gene expression agreed with that of ELISA.
Subject(s)
Humans , Blotting, Northern , Dactinomycin , Enzyme-Linked Immunosorbent Assay , Gene Expression , Hot Temperature , Interleukin-6 , Lymphocytes , Polymerase Chain Reaction , Reverse Transcription , RNA, Messenger , Yeasts , ZymosanABSTRACT
Hairy cell (HC) transformation of human peripheral blood lymphocytes (PBL) by Coxiella burnetii was studied to clarify the significance of persistency of C. burnetii in a hairy cell line (designated "TOL"). TOL cells which exhibited HC characteristics in hairy cell leukemia (HCL) were persistently infected with C. burnetii. Two strains of C. burnetii, our isolate from TOL cells and the original isolate in 1935, the Nine Mile strain from American Type Culture Collection (ATCC, U.S.A), were inoculated to PBL cultures. HC transformation not only by our isolates (87%) but also by Nine Mile strain (100%) was demonstrated in an average of 20 days. The original observation that Coxiella induced HC transformation in vitro was also confirmed in experiments with PBL exposed to C. burnetii in vivo. Spontaneous development of HC were observed in cultures of PBL only from coxiellemic cases (12/24) but not from C. burnetii negative cases (0/57). All HC cell lines (34) as determined by their morphology and cytochemical markers of HC in HCL remained infected with C. burnetii invariably.
Subject(s)
Humans , Blood Cells/microbiology , Cell Line , Cell Transformation, Neoplastic , Coxiella burnetii/isolation & purification , Leukemia, Hairy Cell/microbiology , Lymphocytes/microbiology , Microscopy, Electron, ScanningABSTRACT
Objective:To study the regulation effect of water extraction from Ganoderma Lucidum (Lyess. ex Fr) karst (GL W) on human immune cells and cytokine. Methods: With the technologies of MTT and FACS, the proliferation of T lymphocyte subgroup and IL 6 production in human peripheral blood lymphocytes (PBL) were studied. Results: 1. In the absence of mitogin, GL W could induce not only the inactive lymphocyte but also the PHA activited lymphocyte to proliferate in the similar concentration. The proliferation in the later was more obvious ( P
ABSTRACT
Objective To explore the activation and apoptosis of peripheral blood lymphocytes(PBLs) in children with Henoch-Schonlein purpura(HSP) and the effects of triptoIide(TP) on them. Methods The changes of activation and apoptosis were observed on cultured PBLs in children with HSP and healthy controls ,and the effects of TP were compared respectively. Expression of CD3, CD25 and apoptosis rate of PBLs were assayed with flow cytometry. Results The percentage of CD3+ CD25+ cell was significantly higher (P
ABSTRACT
Gamma interferon (gamma-IFN), a lymphokine produced by activated T lymphocytes, has a variety of effects on target cell. It induces class II antigens of the major histocompatibility complex not only in immunocompetent cells but also in non-immunocompetent cells. gamma-IFN also can exert, in addition to anti-viral activity, a series of anticellular effects on a variety of cell types. The effects of gamma-IFN on the proliferation of cultured epidermal cell (EC) and induction of HLA-DR antigen expression by EC (HLA-DR+KC) were studied. Furthermore, the immunologic role of HLA-DR+KC in the mixed epidermal cell-lymphocyte reaction (MECLR) was studied. The antiproliferative effect of gamma-IFN on the cultured EC was seen 3 days after treatment of gamma-IFN and the effect was dose-dependent. Number of HLA-DR+KC was increased dose-dependently with treatment of gamma-IFN. In MECLR, HLA-DR+KC had been found to exert stimulatory role on allogenic lymphocytes. However, there was no significant role of HLA-DR+KC on autologous lymphocytes.