Apoptosis In Peripheral Blood Lymphocytes Research Articles

The gut microbiota contributes to the infection of bovine viral diarrhea virus in mice.

Bovine viral diarrhea virus (BVDV) is prevalent worldwide and causes significant economic losses. Gut microbiota is a large microbial community and has a variety of biological functions. However, whether there is a correlation between gut microbiota and BVDV infection and what kind of relation between them have not been reported. Here, we found that gut microbiota composition changed in normal mice after infecting with BVDV, but mainly the low abundance microbe was affected. Interestingly, BVDV infection significantly reduced the diversity of gut microbiota and changed its composition in gut microbiota-dysbiosis mice. Furthermore, compared with normal mice of BVDV infection, there were more viral loads in the duodenum, jejunum, spleen, and liver of the gut microbiota-dysbiosis mice. However, feces microbiota transplantation (FMT) reversed these effects. The data above indicated that the dysbiosis of gut microbiota was a key factor in the high infection rate of BVDV. It is found that the IFN-I signal was involved by investigating the underlying mechanisms. The inhibition of the proliferation and increase in the apoptosis of peripheral blood lymphocytes (PBL) were also observed. However, FMT treatment reversed these changes by regulating PI3K/Akt, ERK, and Caspase-9/Caspase-3 pathways. Furthermore, the involvement of butyrate in the pathogenesis of BVDV was also further confirmed. Our results showed for the first time that gut microbiota acts as a key endogenous defense mechanism against BVDV infection; moreover, targeting regulation of gut microbiota structure and abundance may serve as a new strategy to prevent and control the disease.IMPORTANCEWhether the high infection rate of BVDV is related to gut microbiota has not been reported. In addition, most studies on BVDV focus on in vitro experiments, which limits the study of its prevention and control strategy and its pathogenic mechanism. In this study, we successfully confirmed the causal relationship between gut microbiota and BVDV infection as well as the potential molecular mechanism based on a mouse model of BVDV infection and a mouse model of gut microbiota dysbiosis. Meanwhile, a mouse model which is more susceptible to BVDV provided in this study lays an important foundation for further research on prevention and control strategy of BVDV and its pathogenesis. In addition, the antiviral effect of butyrate, the metabolites of butyrate-producing bacteria, has been further revealed. Overall, our findings provide a promising prevention and control strategy to treat this infectious disease which is distributed worldwide.

Lymphocyte apoptosis and its association with the inflammatory markers and disease severity in juvenile-onset systemic lupus erythematosus patients

BackgroundThe defective clearance of apoptotic bodies in juvenile-onset systemic lupus erythematosus (jSLE) potentially leads to the persistence of autoreactive lymphocytes and the perpetuation of the autoimmune response. These factors contribute to the disturbance in lymphocyte apoptosis and show potential as key determinants in the clinical course and severity of jSLE. This study evaluates the role of peripheral blood (PB) lymphocyte apoptosis in prognosis of jSLE and as a predictor for disease activity.MethodsThe study involved 100 jSLE patients and 50 healthy controls. Flow cytometry was used to analyze percentages of lymphocyte apoptosis in PB of all study participants. Plasma levels of pro-inflammatory cytokines were determined using ELISA.ResultsOur results showed that percentages of lymphocyte apoptosis in PB of jSLE patients are significantly higher than those of healthy controls. These percentages are significantly positively associated with disease activity of patients (SLEDAI-2 K). Furthermore, plasma cytokine levels (IL-17, IFN-γ and TNF-α) are significantly elevated in jSLE patients compared to their levels in healthy controls. Also, there are weak significant positive correlations between percentages of PB lymphocyte apoptosis and each of IL-17 and IFN-γ plasma levels in jSLE patients. Moreover, PB lymphocyte apoptosis percentages among jSLE patients are higher in the presence of some clinical and laboratory features than those in their absence.ConclusionPeripheral apoptotic lymphocytes could contribute to the prognosis of jSLE and could be used as a predictor for disease activity in jSLE patients.

The Intensity of Blood Lymphocyte Apoptosis in Chronically Exposed Individuals with Obligate Forms of Precancerous Conditions.

We studied the apoptotic death of peripheral blood lymphocytes in individuals chronically exposed to radiation with a history of obligate forms of precancerous conditions. The study included 242 subjects chronically exposed to low-intensity radiation in the range of low and medium doses (mean dose of irradiation of the red bone marrow 555±39 mGy). Of these, 121 subjects had obligate forms of precancerous conditions. In irradiated individuals with obligate forms of precancers, a statistically significant increase in the frequency of apoptosis of peripheral blood lymphocytes at an early and late stages was revealed. Correlation analysis revealed no dependence of the apoptotic death of peripheral blood lymphocytes on the dose of red bone marrow irradiation.

Pseudomonas aeruginosa quorum-sensing molecule N-(3-oxododecanoyl)-L-homoserine lactone mediates Ca+2 dysregulation, mitochondrial dysfunction, and apoptosis in human peripheral blood lymphocytes

N-(3-oxododecanoyl)-l-homoserine lactone is a Pseudomonas aeruginosa secreted quorum-sensing molecule that mediates the secretion of virulence factors, biofilm formation and plays a pivotal role in proliferation and persistence in the host. Apart from regulating quorum-sensing, the autoinducer signal molecule N-(3-oxododecanoyl)-l-homoserine lactone (3O–C12-HSL or C12) of a LasI–LasR circuit exhibits immunomodulatory effects and induces apoptosis in various host cells. However, the precise pathophysiological impact of C12 on human peripheral blood lymphocytes and its involvement in mitochondrial dysfunction remained largely elusive. In this study, the results suggest that C12 (100 μM) induces upregulation of cytosolic and mitochondrial Ca+2 levels and triggers mitochondrial dysfunction through the generation of mitochondrial ROS (mROS), disruption of mitochondrial transmembrane potential (ΔΨm), and opening of the mitochondrial permeability transition pore (mPTP). Additionally, it was observed that C12 induces phosphatidylserine (PS) exposure and promotes apoptosis in human peripheral blood lymphocytes. However, apoptosis plays a critical role in the homeostasis and development of lymphocytes, whereas enhanced apoptosis can cause immunodeficiency through cell loss. These findings suggest that C12 exerts a detrimental effect on lymphocytes by mediating mitochondrial dysfunction and enhancing apoptosis, which might further impair the effective mounting of immune responses during Pseudomonas aeruginosa-associated infections.

Peripheral blood lymphocytes apoptosis role in rheumatoid arthritis progressing

Rheumatoid arthritis (RA) is an autoimmune, chronic, and genetically linked inflammatory lesion of joint tissues that is accompanied by extra-articular systemic pathologies. The disease progression leads to joints immobilization, and eventually, the patient's disability occurs approximately ten years from the first clinical manifestation. RA pathogenesis involves various mechanisms: specific joint-related damage, nonspecific adaptive, and vessel-related pathological changes. Our research aimed to study the role of peripheral blood lymphocyte apoptosis in RA pathogenesis. We have analyzed research data from Google Scholar, PubMed, Web of Science, and Scopus databases to investigate the role of lymphocyte apoptosis in RA progression. Clinical manifestations in RA are caused by autoreactive T- and B-lymphocyte activity supported by humoral and cellular immune factors activity. Disease pathogenesis is caused by an imbalance in the process of programmed cell death (apoptosis): a proportion of immune cells are rapidly destroyed. In contrast, apoptosis is inhibited in the other classes of immune cells. High infiltration of the joint by autoreactive sensitized lymphocytes worsens the patient's condition. Apoptosis inhibition is especially noticeable in the early stages of RA and correlates with the concentration of the anti-apoptotic molecule Bcl-2 in the synovia. Activating the apoptotic destruction of lymphocytes (by drug action) allows a positive therapeutic effect and sustained remission. However, it should be noted that genetic factors play a significant role in the onset, progression and drug response of RA. In addition, environmental and behavioral factors can activate RA progression and influence treatment efficacy.

Effect of propofol on lymphocyte subsets and postoperative cognitive function in lung cancer model rats through JAK/STAT signaling pathway

With changes in people’s living environment, the incidence of lung cancer is on the rise, seriously affecting people’s physical and mental health. Previous studies have shown that, propofol is used for treating lung cancer. Whether propofol affects lymphocyte subsets and postoperative cognitive function in rats with lung cancer remains unclear. In this study, we established and intervened rat lung cancer model, followed by Flow Cytometry/financal capacity model (FCM) analysis of peripheral blood lymphocyte subsets and apoptosis level. Morris water maze method was used to test cognitive function; while CCK-8 method and Transwell assessed tumor cells activities. qRT-PCR (Quantitative Real-time Polymerase Chain Reaction) and Western blot detected the expression of genes and proteins, respectively. Lung tissue of rats was swollen and alveolar damage was more serious, with tumor cell proliferation in high-dose propofol group being significantly inhibited with reduced migration ability and increased apoptosis. Moreover, Bcl-2 level in the JAK/STAT inhibitor group was lowest, and each lymphocyte subset index increased significantly, while the latency and swimming distance of rats decreased significantly. The expressions of JAK, STAT, and Bcl-2 mRNA in the high-dose propofol+JAK/STAT agonist group were significantly reduced. These expressions were directly regulated by propofol in the higher than high-dose propofol group (P <0.05), suggesting that propofol can directly regulate JAK, STAT, and Bcl-2. Propofol thus improves the lung cancer rats by inhibiting JAK/STAT signaling and down-regulating Bcl-2.

Regulation of kisspeptin-54 activity of indolamine-2,3-dioxygenase and apoptosis of peripheral blood lymphocytes

Pregnancy is a phenomenon of natural semi-allogeneic transplantation, since the fetus is half alien due to the expression of paternal antigens. It was found that the hypothalamic hormone kisspeptin during pregnancy is produced by the syncytiotrophoblast of the placenta and participates in the formation of a new specific hormonal background. Several forms of the hormone circulate in the blood of pregnant women: kisspeptin-10, kisspeptin-14 and kisspeptin-54 (according to the number of amino acid residues in the hormone molecule), but the main active form is kisspeptin-54. The main mechanism for the formation of immune tolerance during pregnancy is the induction of the expression of the enzyme indolamine-2,3- dioxygenase (IDO) by antigen-presenting cells of peripheral blood, resulting in the catalysis of tryptophan (Trp) to kynurenins (KYN) blocking the activation and causing apoptosis of cytotoxic CD8+T lymphocytes in the zone of contact of maternal immune cells with placental-fetal complex antigens. In addition, during pregnancy, an important role is assigned to the process of apoptosis, since activated cells can be potentially dangerous for the developing fetus. Immunocompetent blood cells express a specific membrane receptor of kisspeptin (KISS-1R). Since kisspeptin-54 enters the systemic circulation only during pregnancy, the hormone has an effect on immune cells only during this period.The aim of this work was to evaluate the effect of kisspeptin-54 in concentrations comparable to its levelduring physiological pregnancy on IDO activity and apoptosis of peripheral blood lymphocytes.Peripheral blood mononuclear cells (PBMC) obtained from 10 healthy non-pregnant women of reproductive age (from 23 to 32 years) were used as the object of the study. Lymphocyte apoptosis was assessed in PBMC suspension by staining with annexin-V and propidium iodide. The determination of the number of cells in the early and late stages of apoptosis was carried out in the isolated gate of lymphocytes. IDO activity in PBMC was determined spectrophotometrically by changes in the concentration of KYN, the first stable metabolite of the Trp decay pathway.It was found that kisspeptin-54 at a concentration of 4.6 pM corresponding to the second trimester of pregnancy significantly enhances the activity of IDO, increases the number of cells in the early and late stages of apoptosis. Thus, kisspeptin-54 is an important mechanism for controlling these processes during pregnancy, aimed at protecting the semi-allogeneic fetus from adverse immune reactions of the mother and the favorable development of pregnancy.

Crosstalk between Apoptosis and Cytotoxic Lymphocytes (CTLs) in the Course of Lagovirus Europaeus GI.1a Infection in Rabbits.

Lagovirus europaeus is a single-stranded RNA virus causing an acute fatal disease in wild and domestic rabbits around the world. Studies have shown that the pivotal process impacting the immune response against the disease is apoptosis, registered mainly in hepatocytes and in peripheral blood, together with an increased number of cytotoxic lymphocytes (CTLs). It is known that cytotoxic lymphocytes can induce target cells to undergo apoptosis on the pseudoreceptor pathway, such apoptosis having been found in several acute and chronic viral infections. The study aimed to assess the crosstalk between the apoptosis of peripheral blood lymphocytes and CD8+ T lymphocytes (as CTLs) in rabbits infected with 6 Lagovirus europaeus GI.1a viruses. Sixty rabbits of Polish hybrid breed comprising both sexes and weighing 3.2-4.2 kg were the experimental group, and an identical group was the control. Each of the six GI.1a Lagovirus europaeus viruses was inoculated into ten experimental rabbits. Control rabbits received glycerol as a placebo. Flow cytometric analysis was performed on blood from the study and control group animals for peripheral blood lymphocyte apoptosis and CTL percentage determination. The activation of apoptosis in peripheral blood lymphocytes was recorded from 4 h post inoculation (p.i.) up to 36 h p.i. The percentage of CTLs in the total blood pool decreased from 8 to 36 h p.i. A negative correlation between apoptosis of lymphocytes and the number of CTLs was proven. This may be the first evidence of virus-induced CTL apoptosis in Lagovirus europaeus GI.1a infection.

Molybdenum-Induced Apoptosis of Splenocytes and Thymocytes and Changes of Peripheral Blood in Sheep.

To investigate the effects of molybdenum (Mo) on apoptosis of lymphocytes and changes of peripheral blood in sheep, a total of 20 5-month-old healthy female sheep were randomly divided into five groups of 4 and orally administered with water containing Na2MoO4·2H2O (0, 5, 10, 20, and 50mg/kg BW/day) for 28days. Jugular vein blood was taken on the 0th, 7th, 14th, 21st, and 28th day of Mo treatment, respectively. On the 28th day, the spleen and thymus were removed for observing histopathology and apoptosis-related DNA damage by hematoxylin and eosin (HE) staining and TdT‑mediated dUTP Nick-End Labeling (TUNEL) staining, respectively. The blood routine indexes were determined by an automatic blood analyzer. Further, the apoptosis of lymphocytes and changes in mitochondrial membrane potential (MMP) of peripheral blood were analyzed by flow cytometry. Results showed that excessive Mo induced apoptosis-related DNA damage in the splenocytes and thymocytes and significantly increased the apoptosis indexes of the splenocytes and thymocytes (P < 0.01). Furthermore, the treatment with excessive Mo significantly decreased the MMP (P < 0.01) and promoted apoptosis in peripheral blood lymphocytes (P < 0.01). And the number of WBC, Lymph, Gran, and RBC and the indexes of HGB and HCT were also significantly decreased (P < 0.05 or P < 0.01), while RDW was significantly increased by excessive Mo (P < 0.05 or P < 0.01). In conclusion, excessive Mo-induced DNA damage and apoptosis of the lymphocytes changed the RBC-related indexes of the peripheral blood in sheep.

INTENSITY OF OXIDATIVE PROCESSES IN BLOOD AND LEVEL OF APOPTOSIS IN BLOOD LYMPHOCYTES IN RADIOLOGISTS/X-RAY TECHNOLOGIES EXPOSED TO SMALL DOSES OF IONIZING RADIATION.

to determine the presence of changes in peripheral blood parameters, characterizing its redox state, and the level of apoptosis of lymphocyte in radiologists/x-ray technologies who, due to their official duties, are exposed to small doses of ionizing radiation. The work was performed on blood samples of 45 professionals radiologists/x-ray technologies and 52 conventionally healthy individuals (control group). The content of malondialdehyde and sulfhydryl groups of proteins and peptides (-SH) in blood plasma was determined; catalase enzyme activity and the ratio of pro-antioxidant processes in hemolysates, the level of superoxide anion-radical (О2-) generation, the total production offree radical compounds (reactive forms of oxygen and nitrogen) and the level of apoptosis of peripheral blood lymphocytes (PBL). The content of malondialdehyde in the blood of professionals was increased by 1.49 times and the contentof -SH was decreased by 1.67 times compared to conventionally healthy individuals. An increase in the level of О2-production by 1.56 times was observed for PBL. The obtained results indicate a shift in the ratio between antioxidant and pro-oxidant processes towards the latter, which is confirmed by a 1.49-fold increase of this index. The levelfor PBL apoptosis was reduced by 1.35 times. For professionals, against the background of increased generation of О2-, a reliable direct correlation was observed between the indicator of apoptosis and the total production of free radical compounds, and between the latter and the level of apoptosis of lymphocytes, which was not noted for the conventionally healthy individuals group. A change in the ratio between pro- and antioxidant processes in the blood was found for professionals who are in contact with sources of ionizing radiation, which indicates the possibility of the development of oxidative stress, and the consequence of a reduced level of apoptosis of lymphocytes may be the danger of accumulating genetic damage in these cells.

Methylation Status of Apoptosis Genes and Intensity of Apoptotic Death of Peripheral Blood Lymphocytes in Persons Chronically Exposed to Radiation

Methylation of the CpG islands of gene promoter regions is the most common epigenetic modification involved in the regulation of gene expression. A number of studies have shown that ionizing radiation can cause both hyper- and hypomethylation of DNA. Aberrant methylation affects cellular processes and can lead to the development of various pathological states. In the literature, there are few studies on the methylation status of human DNA a long time after radiation exposure. Here, the methylation level of CpG islands of the promoter regions of apoptosis genes (BCL2, ATM, MDM2, CDKN1A, STAT3, and NFKB1), and also its influence on apoptosis of peripheral blood lymphocytes in chronically exposed persons were studied. Residents of the South Ural region who were chronically exposed to radiation (after discharges of radioactive wastes into the Techa river by the "Mayak Production Association" in 1949-1956) were included in the study. It was established that the proportion of individuals with hypermethylated BCL2 gene promoter among the exposed people was statistically significantly higher than in the control group. The percentage of methylation of the ATM gene promoter weakly positively correlated with dose and age characteristics. Differences in the frequency of lymphocyte apoptosis in exposed individuals with a hypo- or hypermethylated ATM gene promoter were also established. The data indicate that, in the long-term, after chronic low intensity radiation exposure at low and medium doses, epigenetic modifications of the genome occur, which are manifested as changes in methylation of promoter regions of BCL2 and ATM genes.

Analysis of annexin V levels and cytokine status in patients with acute myocardial infarction

Aim. Study of annexin V levels and cytokine status in patients with acute myocardial infarction. Materials and methods. We examined 27 patients with Q-forming myocardial infarction who were admitted 12–24 hours after the development of the clinic, and 20 individuals in the control group. The lipid spectrum was determined using an automatic analyzer Humalizer-3000 (Human, Germany). Immunological studies were performed on a flow cytofluorimeter Navios (Beckman Coulter, USA). Lymphocyte subpopulations were evaluated using monoclonal antibodies produced by Immunotech (Coulter Corporation, USA). A set of ANNEXIN V-FITC/7 AAD (Beckman Coulter, USA) was used to isolate cells that had undergone apoptosis. Isolation of peripheral blood mononuclears was performed by centrifugation on a density gradient Ficol-hypac (Pharmacia Fine Chemicals, Sweden), interleukin-1β was determined using the Vector-Best test system (Vector-Best, Russia), and the tumor necrosis factor was determined using the RayBio Human TNF-alpha ELISA Kit (USA). Statistical data processing was performed using the program package Statistica 10.0. Results. In patients with acute myocardial infarction on the background of dyslipidemia (total cholesterol and LDL cholesterol increased, cholesterol-HDL reduced) showed a significant decrease in the total number of T-lymphocytes (СD 3+) (p &lt; 0.05), T-suppressor (СD 8+) (p &lt; 0.01), B-cells number (СD 19+) and the ratio of CD 4+/CD 8+ lymphocytes increased (p &lt; 0.05). In patients with myocardial infarction there was a significant increase of apoptotic cells and рro-inflammatory cytokines involved in apoptosis (IL-1β and TNF-α) in the blood compared with the control. Conclusion. Increased spontaneous apoptosis of peripheral blood lymphocytes in patients with acute myocardial infarction indicates the systemic nature of apoptosis in acute coronary pathology, due to increased annexin-dependent apoptosis of T-lymphocytes as a result of immune disorders, which leads to activation of the cytokine system with an increased concentration of рro-inflammatory mediators (TNF-α, IL-1β). The identified dependencies require further study with the prospect of developing predictive algorithms for myocardial infarction outcomes based on indicators of annexin V and pro-inflammatory cytokines.

Evaluation of the antiviral effect of four plant polysaccharides against duck circovirus

Recently, outbreaks of duck circovirus (DuCV) are frequently occurring worldwide due to secondary infections caused by post infection-induced immunosuppression. Due to a lack of preventive drugs and vaccines, the waterfowl industry losses are ever increasing. In this study, we extracted Astragalus polysaccharides (APS), pine pollen polysaccharides (PPPS), Aloe vera polysaccharides (AVE), and Ficus carica polysaccharides (FCPS) from Astragalus, pine pollen, aloe, and F. carica leaves, respectively. We randomly divided 150 one-day-old Cherry Valley ducks into five groups, which were inoculated with the DuCV solution and orally administered APS, PPPS, AVE, FCPS, and phosphate buffer saline (PBS), respectively. We collected the duck immune organs and serum samples at 8, 16, 24, 32, 40, and 48 days post-infection (dpi). Using clinical symptom analysis, molecular biology experiments, and serological experiments, we proved that plant polysaccharides could (a) improve the duck immunity, (b) reduce the viral load, and (c) mitigate DuCV-induced damage to immune organs, with both APS and PPPS having significant effects. Moreover, we detected viral load and cytokines within the first 8 dpi. Since the body's innate immunity could inhibit viral replication within five days of virus infection, 1–5 dpi was the best treatment time. Among the four polysaccharides showing in vitro anti-apoptotic activity, APS and PPPS significantly inhibited the DuCV infection-induced apoptosis of peripheral blood lymphocytes. Overall, since our findings show APS and PPPS having significant anti-DuCV effects both in vivo and in vitro, they can be promising candidates for preventing DuCV infection in ducks.

Complex molecular mechanism of ammonia-induced apoptosis in chicken peripheral blood lymphocytes: miR-27b-3p, heat shock proteins, immunosuppression, death receptor pathway, and mitochondrial pathway

Ammonia gas, a toxic environmental pollutant, is a vital component of PM2.5 aerosols, and can decrease human and animal immunity. Peripheral blood lymphocytes (PBLs) are main immune cells. Nevertheless, poisoning mechanism of PBLs under ammonia exposure remains unclear. Here, we established an ammonia poisoning model of chicken PBLs to explore poisoning mechanism of ammonia-caused apoptosis in chicken PBLs. Cell viability and apoptosis rate were detected using CCK8 assay and flow cytometry, respectively. Mitochondrial membrane potential (MMP) was observed using fluorescent staining. In addition, qRT-PCR was performed to measure mRNA levels of apoptosis-related genes (tumor necrosis factor-α (TNF-α), tumor necrosis factor receptor 1 (TNFR1), TNF receptor-associated death domain (TRADD), Fas-associated death domain (FADD), Caspase-8, BH3-interacting domain death agonist (Bid), Bcl-2-associated X protein (Bax), Bcl-2 homologous antagonist/killer (Bak), B-cell lymphoma-2 (Bcl-2), Cytochrome-c (Cytc), apoptotic protease activating factor-1 (APAF1), Caspase-9, and Caspase-3), immune-related genes (interferon-γ (IFN-γ), interleukin-2 (IL-2), IL-4, IL-6, IL-1β, IL-10, transforming growth factor-β1 (TGF-β1), IL-17, IL-21, and IL-22), heat shock protein (HSP) genes (HSP25, HSP40, HSP60, HSP70, HSP90, and HSP110), as well as miR-27b-3p. Western blot was used to determine protein levels of apoptosis-related factors (TNF-α, Caspase-8, Bcl-2, Caspase-9, and Caspase-3), as well as HSPs (HSP40, HSP60, HSP70, and HSP90). The results indicated that TRADD, FADD, and APAF1 were target genes of miR-27b-3p, as well as miR-27b-3p participated in molecular mechanism of apoptosis through targeting TNF-α/TNFR1/Caspase-8 death receptor pathway-triggered Bid/Cytc/Caspase-9 mitochondrial pathway in ammonia-treated chicken PBLs. In addition, our findings demonstrated that excess ammonia led to immunosuppression via Th1/Th2 imbalance and Treg/Th17 imbalance. Simultaneously, ammonia stress activated HSPs. In summary, for the first time, our data demonstrated that HSPs-triggered immunosuppression led to apoptosis under ammonia exposure. Our findings provided a new insight into molecular mechanism of ammonia poisoning and an important reference for environmental risk assessment related to ammonia.

Transcriptional activity of repair, apoptosis and cell cycle genes (TP53, MDM2, ATM, BAX, BCL-2, CDKN1A, OGG1, XPC, PADI4, MAPK8, NF-KB1, STAT3, GATA3) in chronically exposed persons with different intensity of apoptosis of peripheral blood lymphocytes.

Transcriptional activity of genes involved in maintaining genetic homeostasis (genes for repair, cell cycle and apoptosis: TP53, MDM2, ATM, BAX, BCL-2, CDKN1A, OGG1, XPC, PADI4, MAPK8, NF-KB1, STAT3, GATA3) was studied in chronically exposed persons with an increased intensity of early and late stages of apoptosis and necrosis of peripheral blood lymphocytes. The object of this study was peripheral blood mononuclear cells obtained from 132 chronically exposed residents of the Techa riverside villages. The mean accumulated dose to red bone marrow was 426.4 ± 48.2 mGy (1.3–2930.0 mGy), to thymus and peripheral immune organs, 58.9 ± 7.9 mGy (0.1–489.0 mGy). The study was performed more than 60 years after the onset of exposure, the average age of exposed persons was 68 ± 0.6 years (55–86 years). The study of apoptotic and necrotic death of peripheral blood lymphocytes was based on the presence of phosphatidylserine on the cell membrane surface, as well as on its permeability for DNA-intercalating dye. Evaluation of the relative content of mRNA genes for repair, cell cycle, and apoptosis was carried out using real-time PCR. An increased relative content of PADI4 gene mRNA was registered in the group of chronically exposed persons with the increased intensity of early apoptosis (p = 0.006). Modulation of the relative content of mRNA of the TP53 (p = 0.013) and BCL-2 (p = 0.021) genes was detected in the group of chronically exposed individuals with the increased intensity of the late stage of apoptosis. A statistically signif icant increase in the transcriptional activity of the TP53 gene was observed in the group of chronically exposed persons with the increased intensity of peripheral blood lymphocyte necrosis in the long-term period (p = 0.015). In the course of the study it was noted that exposed people with increased intensity of apoptosis, f irst of all, demonstrate changes in the transcriptional activity of apoptotic genes. These data are consistent with current views on the activation of programmed cell death.

On the Question of the Norm of some Laboratory Indicators of Homeostasis in People over 60

Currently, there are no reference values for many laboratory indicators of homeostasis for gerontological patients, which complicates the objective interpretation of their disorders.Aim of study. Based on a comparison of the reference values of some laboratory indicators of homeostasis of volunteers over 65 and people of working age, we offer their conditional norm for people of gerontological age.Materials and methods. Studies of laboratory indicators of homeostasis were carried out in 25 volunteers aged 60 to 85 years. The comparison group consisted of 50 donors aged 18–59 years. Investigated indicators: lipid peroxidation and antioxidant blood system; factors of endogenous vascular regulation; apoptosis of peripheral blood lymphocytes; blood rheology; endotoxemia, immunology. Statistical analysis of the data was performed using the Statistica 10 software package (StatSoft, Inc., USA); when comparing the indicators, the nonparametric Mann–Whitney U test (abnormal distribution) was used.Results. In elderly and senile people, due to physiological aging, as well as the influence of endogenous and exogenous factors and concomitant diseases, there are significant differences in the reference values of some laboratory parameters from the parameters of the same name for people of working age.

Controlled cell deletion of peripheral blood lymphocytes in type 2 diabetes mellitus

Background: Controlled cell deletion (apoptosis) and cellular proliferation are the main mechanisms that are responsible for shaping tissues and organs in developing embryos. During adult life, apoptosis serves a protective mechanism which eliminates old, useless, and damaged cells. In healthy organisms apoptosis and cell proliferation are in balance. Aim of the study: To detect apoptosis of peripheral blood lymphocytes in type 2 diabetes mellitus patients and its relation with the percentage of glycohemoglobin and the effect on the outcome of the disease. Method: This study was carried out on 40 volunteers, twenty of them (9 males and 11 females) were patients with type 2 diabetes and twenty (5 males and 15 females) were normal healthy subjects and was served as controls of the study. From each patient a sample of blood was aspirated for detection of peripheral blood lymphocyte apoptosis and for the measurement of the percentage of glycated hemoglobin (HbA1c). For the control subjects, blood samples were aspirated to detect peripheral blood lymphocyte apoptosis. Results: The percentage of peripheral blood lymphocytes apoptosis increased significantly (P

PD-1 Blockade Restores the Proliferation of Peripheral Blood Lymphocyte and Inhibits Lymphocyte Apoptosis in a BALB/c Mouse Model of CP BVDV Acute Infection.

Acute infection of bovine viral diarrhea virus (BVDV) is associated with immune dysfunction and can cause peripheral blood lymphopenia and lymphocyte apoptosis. Our previous study has confirmed that programmed death-1 (PD-1) blockade inhibits peripheral blood lymphocytes (PBL) apoptosis and restores proliferation and anti-viral immune functions of lymphocytes after BVDV infection in vitro. However, the situation in vivo remains to be further studied and confirmed. Therefore, in this study, we established a BALB/c mouse model of acute BVDV infection with cytopathic (CP) BVDV (strain NADL) and non-cytopathic (NCP) BVDV (strain NY-1). Then, we examined the mRNA and protein levels of PD-1 and programmed death-ligand 1 (PD-L1) in peripheral blood mononuclear cells (PBMC) from BVDV-infected mice and analyzed the effects of PD-1 blockade on the proportions of CD3+, CD4+, and CD8+ T cell subsets, the apoptosis and proliferation of PBL, and the production of IL-2 and IFN-γ. We found that leukopenia, lymphocytopenia, and thrombocytopenia were developed in both CP and NCP BVDV-infected mice at day 7 of post-infection. The mRNA and protein expression of PD-1 and PD-L1 were significantly upregulated in CP and NCP BVDV-infected mice. Moreover, PD-1/PD-L1 upregulation was accompanied by leukopenia and lymphopenia. Additionally, PD-1 blockade inhibited PBL apoptosis and virus replication, restored the proportions of CD3+, CD4+, and CD8+ T cell subsets, and increased IFN-γ production and p-ERK expression in BVDV-infected mice. However, blocking PD-1 did not significantly affect PBL proliferation and IL-2 production in NCP BVDV-infected mice. Our findings further confirmed the immunomodulatory role of PD-1 in peripheral blood lymphocytopenia in vivo and provided a scientific basis for exploring the molecular mechanism of immune dysfunction caused by acute BVDV infection.

Effects of sevoflurane exposure on apoptosis and cell cycle of peripheral blood lymphocytes, and immunologic function

BackgroundWaste anesthetic gases (WAGs) leaked from new-type halogenated inhalational anesthetics such as sevoflurane have been were reported to pose a risk for the health of operating room personnel. The effects of WAGs on peripheral blood lymphocytes, however, remain yet controversial. The present study was undertaken to examine the effects of occupational sevoflurane exposure on the peripheral blood lymphocytes of medical personnel who work in the operating room.MethodsA cohort of 56 medical residents were divided into exposed group (n = 28) and control group (non-exposed group) (n = 28). Gas chromatography was used to measure the concentration of sevoflurane in the medical resident’s breathing zone during surgeries under inhalation anesthesia in the exposure group. The gas collection lasted an hour. Peripheral blood lymphocytes were isolated from venous blood, and then apoptosis and cell cycle were analyzed by flow cytometry. EDTA-anticoagulated whole blood was harvested to analyze the lymphocyte subsets by flow cytometry. Immunoglobulins (IgA, IgM, IgG) were quantified by immunoturbidimetry.ResultsThe average concentration of sevoflurane in the exposed group was 1.03 ppm with a range from 0.03 ppm to 2.24 ppm. No significant effects were found on the apoptosis rates or cell cycles of peripheral blood lymphocytes in the exposed group relative to the control group (P > 0.05). Similarly, there were no significant differences in the lymphocyte subsets or the levels of immunoglobulins (IgA, IgM, IgG) between the two groups (P > 0.05).ConclusionsOccupational exposure to low-level sevoflurane has no significant effect on the peripheral blood lymphocytes of operating room staff, but this conclusion needs to be confirmed by multicenter and long-term follow-up studies with large samples.Trial registration number and date of registrationChiCTR2000040772, December 9, 2020 (Retrospective registration).

Altered VDAC-HK association and apoptosis in mouse peripheral blood lymphocytes exposed to diabetic condition: an in vitro and in vivo study

Increased apoptotic lymphocytes have been correlated to a high incidence of infection in poorly controlled diabetes. This study aimed to determine whether altered voltage-dependent anion channel (VDAC)-hexokinase (HK) association contributes to the increase in apoptosis. Mouse peripheral blood lymphocytes (PBL) exposed to high glucose (Glc)/palmitic acid (PA) were used as the in vitro model, which was compared with PBL isolated from alloxan-induced diabetic mice (in vivo model). Our results showed a significant increase in apoptosis as indicated by the apoptotic index, caspase-3 activity, mitochondrial membrane potential and ultrastructural study. HK and glucose-6-phosphate dehydrogenase (G6PDH) activities were markedly reduced with a profound increase in glucose-6-phosphate level. Co-immunoprecipitation confirms HK interaction with VDAC, an outer mitochondrial membrane protein. Inhibited glycolytic enzyme, i.e. HK and reduced HK-VDAC interaction in our study could contribute to increased apoptosis in lymphocytes exposed to high Glc/PA. Targeting HK-VDAC interaction may therefore provide therapeutic potential for the treatment of diabetes-associated infection.