Lymphocyte Cell Cycle Research Articles

Galectin-1-producing mesenchymal stem cells restrain the proliferation of T lymphocytes from patients with systemic lupus erythematosus

Introduction Bone marrow mesenchymal stem cell (BMMSC) transplantation is beneficial in treating Systemic lupus erythematosus (SLE); however, the underlying mechanism remains elusive. This study investigates the role of BMMSCs in regulating lymphocyte proliferation and cell cycle progression during SLE and delves into the contribution of BMMSC-produced galectin-1. Methods BMMSCs were co-cultured with T lymphocytes to assess their impact on suppressing CD4+ T cells in SLE patients. Proliferation and cell cycle distribution of CD4+ T cells were analyzed using flow cytometry. The expression of cell cycle-related proteins, including p21, p27, and cyclin-dependent kinase 2 (CDK2), was investigated through western blotting. Extracellular and intracellular galectin-1 levels were determined via ELISA and flow cytometry. The role of galectin-1 in CD4+ T cell proliferation and cell cycle was evaluated through RNAi-mediated galectin-1 expression disruption in BMMSCs. Results and discussion BMMSCs effectively inhibited CD4+ T cell proliferation and impeded their cell cycle progression in SLE patients, concurrently resulting in a reduction in CDK2 levels and an increase in p21 and p27 expression. Moreover, BMMSCs expressed a high level of galectin-1 in the co-culture system. Galectin-1 was found to be critical in maintaining the suppressive activity of BMMSCs and restoring the cell cycle of CD4+ T cells. Conclusion This study demonstrates that BMMSCs suppress the proliferation and influence the cell cycle of CD4+ T cells in SLE patients, an effect mediated by the upregulation of galectin-1 in BMMSCs.

Sinomenine Hydrochloride Protects IgA Nephropathy Through Regulating Cell Growth and Apoptosis of T and B Lymphocytes.

Sinomenine hydrochloride (SH) is used to treat chronic inflammatory diseases such as rheumatoid arthritis and may also be efficacious against Immunoglobulin A nephropathy (IgAN). However, no trial has investigated the molecular mechanism of SH on IgAN. Therefore, this study aims to investigate the effect and mechanism of SH on IgAN. The pathological changes and IgA and C3 depositions in the kidney of an IgAN rat model were detected by periodic acid-Schiff (PAS) and direct immunofluorescence staining. After extracting T and B cells using immunomagnetic beads, we assessed their purity, cell cycle phase, and apoptosis stage through flow cytometry. Furthermore, we quantified cell cycle-related and apoptosis-associated proteins by Western blotting. SH reduced IgA and C3 depositions in stage 4 IgAN, thereby decreasing inflammatory cellular infiltration and mesangial injury in an IgAN model induced using heteroproteins. Furthermore, SH arrested the cell cycle of lymphocytes T and B from the spleen of IgAN rats. Regarding the mechanism, our results demonstrated that SH regulated the Cyclin D1 and Cyclin E1 protein levels for arresting the cell cycle and it also regulated Bax and Bcl-2 protein levels, thus increasing Cleaved caspase-3 protein levels in Jurkat T and Ramos B cells. SH exerts a dual regulation on the cell cycle and apoptosis of T and B cells by controlling cell cycle-related and apoptosis-associated proteins; it also reduces inflammatory cellular infiltration and mesangial proliferation. These are the major mechanisms of SH in IgAN.

Homozygous DBF4 mutation as a cause of severe congenital neutropenia

Severe congenital neutropenia presents with recurrent infections early in life as a result of arrested granulopoiesis. Multiple genetic defects are known to block granulocyte differentiation; however, a genetic cause remains unknown in approximately 40% of cases. We aimed to characterize a patient with severe congenital neutropenia and syndromic features without a genetic diagnosis. Whole exome sequencing results were validated using flow cytometry, Western blotting, coimmunoprecipitation, quantitative PCR, cell cycle and proliferation analysis of lymphocytes and fibroblasts and granulocytic differentiation of primary CD34+ and HL-60 cells. We identified a homozygous missense mutation in DBF4 in a patient with mild extra-uterine growth retardation, facial dysmorphism and severe congenital neutropenia. DBF4 is the regulatory subunit of the CDC7 kinase, together known as DBF4-dependent kinase (DDK), the complex essential for DNA replication initiation. The DBF4 variant demonstrated impaired ability to bind CDC7, resulting in decreased DDK-mediated phosphorylation, defective S-phase entry and progression and impaired differentiation of granulocytes associated with activation of the p53-p21 pathway. The introduction of wild-type DBF4 into patient CD34+ cells rescued the promyelocyte differentiation arrest. Hypomorphic DBF4 mutation causes autosomal-recessive severe congenital neutropenia with syndromic features.

Assessment of the functions of cultured lymphocytes when exposed to drugs used in cosmetology

The aim of the study was to determine the number of lymphocytes, intracellular cytokines produced by lymphocytes, and the cell cycle of lymphocytes isolated from the blood of patients when exposed to various drugs, as well as to assess the functions of cultured lymphocytes when exposed to drugs in vivo and in vitro.Materials and methods. The study involved lymphocytes isolated from the blood of healthy women under various conditions. At the first stage of the study, T-lymphocytes were isolated from the blood of patients before exposure to the drug. The absolute and relative lymphocyte count, the number of intracellular cytokines, and the cell cycle were determined.At the second stage, the drugs were added to the nutrient medium, where lymphocytes isolated from the blood of patients who did not receive systemic drugs were cultured. The placental extract preparation was added to the lymphocytes isolated from the first group of patients, while the hyaluronic acid preparation was added to the lymphocytes isolated from the second group of patients.At the third stage, the lymphocytes isolated from the blood of patients after systemic exposure to the placental extract preparation or hyaluronic acid preparation were isolated and cultured, after which the same lymphocyte parameters were determined.Results. The number of T-lymphocytes increased with the systemic use of the placental extract and hyaluronic acid preparations and practically did not change compared with the baseline data, when these drugs were added to the nutrient medium. Placental extract and hyaluronic acid had a positive effect on the mitotic activity of cells; it is worth noting that the effect of placental extract was greater than that of hyaluronic acid. Both drugs did not have a negative effect on apoptosis of T-lymphocytes. Under the effect of placental extract, lymphocytes secreted more interleukins, which contributed to proliferation of keratinocytes.Conclusion. The placental extract and hyaluronic acid preparations have a stimulating effect on keratinocytes. The placental extract preparation has a stimulating effect on T-lymphocytes after systemic exposure of the body to it.

Traditional Chinese medicine CFF-1 exerts a potent anti-tumor immunity to hinder tumor growth and metastasis in prostate cancer through EGFR/JAK1/STAT3 pathway to inhibit PD-1/PD-L1 checkpoint signaling

BackgroundTraditional Chinese Medicine (TCM) CFF-1 has been used in clinic for prostate cancer therapy in China. We reported before CFF-1 induced cell apoptosis via suppressing EGFR-related pathways, reminding us its potential role associated with antitumor immunity. PurposeThe study was aimed to investigate the regulatory mechanism of CFF-1 on PD-L1/PD-1-mediated tumor immune escape. MethodsProstate-specific antigen (PSA) test and the functional assessment of cancer therapy-prostate (FACT-P) and karnosky performance status (KPS) questionnaires were carried out to evaluate patient’ condition before and after therapy. Flow cytometry (FCM) was used for analyzing cell apoptosis, T lymphocyte subsets and cell cycle. Western blotting and Immunohistochemistry (IHC) were performed to measure protein expressions. The synergy of drug combination was assessed by calculating combination index (CI). ResultsCFF-1 obviously decreased PSA and improved the quality of life in patients with advanced prostate cancer. PD-L1 was highly expressed in prostate cancer cells including LNCaP, 22Rv1, PC-3, DU145 and RM-1. PD-1/PD-L1 was upregulated in tumorigenesis and tumor progression of subcutaneous homograft mouse model with immune response, where CD3+ T cell subsets were declined. CFF-1 inhibited PD-L1 expression in prostate cancer cells in a time/dose-dependent manner and blocked tumor growth by suppressing PD-1/PD-L1 upregulation to promote the recovery of CD3+ T lymphocytes, especially CD4+ T cell subset, accompanied by the downregulation of CD4+ FOXP3+ T cell subset. CFF-1 also prolonged the survival and inhibited lung metastasis in tail vein prostate cancer mouse model while repressing PD-1/PD-L1. CFF-1 in combination with docetaxol (DTX) produced a synergistic effects by sensitizing the inhibitory effect of DTX on JAK1/STAT3 pathway targeting PD-L1 blockade. ConclusionCFF-1 inhibited tumor growth and lung metastasis by blocking PD-1/PD-L1 to ameliorate T lymphocyte immune response through EGFR/JAK1/STAT3 pathway, suggesting that CFF-1 might be a promising treatment to resist tumor immunosuppression for prostate cancer patients.

Human myeloma RPMI8226 cells and normal cells have different sensitivity to bisilicate silver nanoparticles in vitro

Introduction. Silver nanoparticles due to its pronounced cytotoxicity are regarded as promising agent for anticancer therapy. Determination of normal and transformed cells sensitivity to silver nanoparticles can be the basis for the application as an adjuvant cancer treatment. The objective of the study was to investigate influence of atomic clusters of Argentum (ACA) in the form of silver bisilicate nanoparticles colloid solution on viability and proliferation of human myeloma cell line, mesenchymal stromal cells and blood lymphocytes. Material and methods. Cell viability was evaluated by MTT and LDH assay. Cell proliferation was evaluated by flow cytometry. Results. It was found that ACA had dose-depending cytotoxicity toward all investigated cell types, but normal and transformed cells varied significantly in the sensitivity to nanoparticles. IC50 for myeloma cell line RPMI8226 was 1,75 µg/ml. For MSCs of different origin IC50 was in the range of 12 to 16 µg/ml. ACA in concentration from 2 to 3 µg/ml induced RPMI8226 cells metabolic disruption and death without influence on viability and cell cycle of mesenchymal stromal cells and blood lymphocytes. Conclusion. Results of work has shown distinct differences in sensitivity to ACA between myeloma cells, mesenchymal stromal cells and blood lymphocytes. The optimal range of ACA concentration with anticancer effect without cytotoxic influence on normal cells has been determined in vitro.

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).

СHARACTERISTICS OF CYCLIN D1-MEDIATED REGULATION OF CELL CYCLE OF PERIPHERAL BLOOD LIMPHOCYTES OF CHORNOBYL CLEAN-UP WORKERS AND PERSONS WITH MALIGNANT NEOPLASMS OF THE ORAL CAVITY, OROPHARYNX AND LARYNGOPHARYNX.

to explore proliferative potential of peripheral blood lymphocytes of Chornobyl clean-up workers and persons with malignant neoplasms of the oral cavity, oropharynx and laryngopharynx by level of expression of cyclin D1 and quantitative parameters of cell cycle. A total of 294 men aged (58.47 ± 7.32) were surveyed, 215 of them were Chornobyl clean"up workers (1986-1987), exposed at the dose range 10.43-3623.31 mSv; 49 persons of the control group and 30persons with malignant neoplasms of the oral cavity, oropharynx and laryngopharynx at III, IVА and IVВ stages ofthe disease. The analysis of parameters of cell cycle and proliferative activity of peripheral blood (PB) lymphocyteswas performed using the flow cytometry. The evaluation of distribution of cells by G0/G1, S, G2/M cell cycle phaseswas done in vivo and in in vitro. Proliferative potential was analyzed by level of expression of cytoplasmic protein ofcyclin D1. Proliferative potential of PB lymphocytes of Chornobyl clean"up workers and persons with malignant neo"plasms of the oral cavity, oropharynx and laryngopharynx was assessed. An increase in the level of spontaneousсyclin D1 expression and disturbance of сyclin D1-dependent regulation of cell cycle of PB lymphocytes after mito"gen activation were determined in the Chornobyl clean-up workers. An increase in pool of cells in the S" and G2/M"phases of cell cycle was detected, which characterizes high proliferative potential of PB lymphocytes. These changesare most pronounced in the subgroup of persons with a radiation dose of D > 500 mSv, and in persons with oncolo"gical pathology. A positive linear dependence has been established between the radiation dose and the number of cellsin the S"phase of cell cycle in the subgroup of Chornobyl clean"up workers with a radiation dose of D > 500 mSv. The detected changes of cyclin D1-dependent regulation of cell cycle and proliferative status of lymphocytes depend on the radiation dose, can be a manifestation of genome instability and be a cause for risks of oncogenesis, in a remote period after radiation exposure.

Effects of flavonoids from Allium mongolicum Regel on cell apoptosis, cell cycle and STAT1 mRNA expression in peripheral blood lymphocytes of sheep

ABSTRACT Flavonoids from Allium mongolicum Regel (AMR) have immunoregulatory and anti-inflammatory properties. However, the effect of flavonoids from A. mongolicum Regel on sheep peripheral blood lymphocytes is still unknown. In this study, we found that total flavonoids from A. mongolicum Regel reduced the apoptosis of peripheral blood lymphocytes in sheep, increased the cell cycle progression to the G2/M phase, and promoted mitosis. These molecules inhibited mRNA expression of the apoptosis-related gene STAT1, indicating that the flavonoids alter JAK-STAT signalling and through this classical pathway inhibit cell apoptosis. The bioactive effects are predominantly related to the structure of flavonoids from A. mongolicum Regel, and the 55% elution group, which contained rutin, quercetin, and mignonette element -5-0-glucose-4-hydroxybenzoic acid, compounds found in plants, was the most effective. The effects were the strongest at a flavonoid concentration of 100μg/ml. In conclusion, the present study showed that flavonoids from A. mongolicum Regel inhibited the expression of STAT1 mRNA, reduced the apoptosis of sheep peripheral blood lymphocytes and promoted cell cycle progression. They also increased the proportion of cells in the G2/M phase. Our findings confirmed a novel role of flavonoids from A. mongolicum Regel in regulating immune function.

Integrated Transcriptome and Histone Modification Analysis Reveals NDV Infection Under Heat Stress Affects Bursa Development and Proliferation in Susceptible Chicken Line.

Two environmental factors, Newcastle disease and heat stress, are concurrently negatively impacting poultry worldwide and warrant greater attention into developing genetic resistance within chickens. Using two genetically distinct and highly inbred layer lines, Fayoumi and Leghorn, we explored how different genetic backgrounds affect the bursal response to a treatment of simultaneous Newcastle disease virus (NDV) infection at 6 days postinfection (dpi) while under chronic heat stress. The bursa is a primary lymphoid organ within birds and is crucial for the development of B cells. We performed RNA-seq and ChIP-seq targeting histone modifications on bursa tissue. Differential gene expression revealed that Leghorn, compared to Fayoumi, had significant down-regulation in genes involved in cell proliferation, cell cycle, and cell division. Interestingly, we also found greater differences in histone modification levels in response to treatment in Leghorns than Fayoumis, and biological processes enriched in associated target genes of H3K27ac and H3K4me1 were similarly associated with cell cycle and receptor signaling of lymphocytes. Lastly, we found candidate variants between the two genetic lines within exons of differentially expressed genes and regulatory elements with differential histone modification enrichment between the lines, which provides a strong foundation for understanding the effects of genetic variation on NDV resistance under heat stress. This study provides further understanding of the cellular mechanisms affected by NDV infection under heat stress in chicken bursa and identified potential genes and regulatory regions that may be targets for developing genetic resistance within chickens.

CHANGES OF CYCLIN D1-DEPENDENT REGULATION OF CELL CYCLE IN PERIPHERAL BLOOD LYMPHOCYTES OF CHORNOBYL CLEAN-UP WORKERS AT A REMOTE PERIOD AFTER RADIATION EXPOSURE.

To study proliferative potential of peripheral blood lymphocytes of Chornobyl clean-up workers by levelof expression of cyclin D1 and quantitative parameters of cell cycle at a remote period after radiation exposure. The research subject was the peripheral blood lymphocytes (PB) of Chornobyl clean-upworkers 30-33 years after radiation exposure. A total of 207 men were surveyed, 164 of them were clean-up workers exposed in the dose range 10.43-3623.31 mSv and 43 persons of the control group. Analysis of proliferationpotential (cell cycle initiation) and cyclin D1 expression in PB lymphocytes were performed in vitro by a micro methodof whole blood leukocytes culture with phytohemagglutinine-P (PHA). Sample preparation was performed by a standard immunofluorescent assay for intracellular proteins using the FITC labelled Mouse Anti-Human Cyclin D1Antibody Set. Cell distribution by cell cycle phases studied by propidium iodide DNA staining and analysis onFACSCalibur laser flow cytometer in histogram mode with separation of G0/G1-, S- and G2/M-regions and Sub-G0/G1-region (apoptotic cells). An increase in the level of spontaneous сyclin D1 expression and disturbance of сyclinD1-dependent regulation of cell cycle of PB lymphocytes after mitogen activation were determined in a remote period after radiation exposure. An increase in the level of cyclin D1 expression was accompanied by increase in pool ofcells in the S- and G2/M-phases of cell cycle which characterizes the high proliferative potential of PB lymphocytes.Mitogen-induced delay of cell cycle of lymphocytes in G1/S check point and reduction of S-phase was revealed.These changes are a manifestation of genomic instability caused by the effect of radiation and depend on the radiation dose. The results confirm the hypothesis about the significance of levels of cyclin D1 expression, as a criterion for manifestations of genome instability and risks of oncogenesis in a remote period after irradiation.

A Conserved Requirement for Fbxo7 During Male Germ Cell Cytoplasmic Remodeling.

Fbxo7 is the substrate-recognition subunit of an SCF-type ubiquitin E3 ligase complex. It has physiologically important functions in regulating mitophagy, proteasome activity and the cell cycle in multiple cell types, like neurons, lymphocytes and erythrocytes. Here, we show that in addition to the previously known Parkinsonian and hematopoietic phenotypes, male mice with reduced Fbxo7 expression are sterile. In these males, despite successful meiosis, nuclear elongation and eviction of histones from chromatin, the developing spermatids are phagocytosed by Sertoli cells during late spermiogenesis, as the spermatids undergo cytoplasmic remodeling. Surprisingly, despite the loss of all germ cells, there was no evidence of the symplast formation and cell sloughing that is typically associated with spermatid death in other mouse sterility models, suggesting that novel cell death and/or cell disposal mechanisms may be engaged in Fbxo7 mutant males. Mutation of the Drosophila Fbxo7 ortholog, nutcracker (ntc) also leads to sterility with germ cell death during cytoplasmic remodeling, indicating that the requirement for Fbxo7 at this stage is conserved. The ntc phenotype was attributed to decreased levels of the proteasome regulator, DmPI31 and reduced proteasome activity. Consistent with the fly model, we observe a reduction in PI31 levels in mutant mice; however, there is no alteration in proteasome activity in whole mouse testes. Our results are consistent with findings that Fbxo7 regulates PI31 protein levels, and indicates that a defect at the late stages of spermiogenesis, possibly due to faulty spatial dynamics of proteasomes during cytoplasmic remodeling, may underlie the fertility phenotype in mice.

Intrinsic function of STAT3 in B cells is to promote B cell proliferation and maturation

Abstract We previously showed that STAT3 promotes T cell survival by restraining unbridled activation and proliferation of T cells (J Biol Chem. 286:30888–97). We also showed that STAT3 regulates Th17 differentiation and the loss of STAT3 in CD4+ T cells prevents development of experimental autoimmune diseases (J Immunol. 180:6070–6). However, the intrinsic and extrinsic functions of STAT3 in B cells are not well understood. In this study, we have generated mice lacking STAT3 in CD19+ B cells and investigated the functions of STAT3 in B cells. Targeted deletion of stat3 in the conditional STAT3KO mouse strain (CD19-CRE-STAT3KO) was verified by Western blot analysis by showing loss of STAT3 expression in CD19+ B cells. We further show by lymphocyte proliferation assays and cell cycle analysis that the CD19-STAT3KO mice proliferated less compared to WT counterparts. Moreover, compared to the development of WT B cells, the STAT3KO mice contained a significantly higher percentage of immature B cells. These results suggest that STAT3 might exhibit diametrically opposite effects in B and T cells; Whereas STAT3 promotes B cell proliferation, STAT3 promotes T-cell survival and inhibits T cell activation/proliferation through up-regulation of Class O Forkhead Transcription Factors. In summary, our study reveals for the first time that STAT3 is required for B cell maturation.

The immunomodulatory function of human amniotic fluid stromal cells on B lymphocytes

Current treatments for B cell-mediated disease are mainly based on global B cell depletion, thereby eliminating pathogenic B cells as well as Breg subsets. A more refined modulation of B cell activity could prove beneficial for patient treatment.Objective:To investigate the immunomodulatory function of human amniotic fluid stromal cells (hAFSCs) on different subpopulation of B lymphocytes.Methods:hAFSCs were isolated and cultured and identified by characteristic phenotypic markers. After coculture of B lymphocytes with hAFSCs, the activation, proliferation, differentiation, as well as apoptosis, cell cycle, and expression of the inhibitory costimulatory molecules B7H1, B7H3, and B7H4 of B lymphocytes were examined in vitro.Results:Coculture with hAFSCs significantly decreased the expression of CD80/CD86, Ki-67 and CFSE expression, on activated B lymphocytes. These might be due to the inhibition of B lymphocyte apoptosis and cell cycle arrest. In activated B lymphocytes, coculture with hAFSCs resulted in a reduced proportion of memory B and plasma cells, reduced amounts of immunoglobulins. hAFSCs could balance the B1 to B2 cell subpopulation ratio. hAFSCs could inhibit the expression of the negative co-inhibitory molecule B7H4 and PD-L1 on the activated B lymphocytes.Conclusion:hAFSCs could inhibit B cell activation, proliferation, and subpopulation differentiation. These might be due to their affect on B cell apoptosis, cell cycle and the expression of costimulatory molecules of human B lymphocytes. Our experiment provided the evidence for hAFSCs as ideal seed cells with therapeutic potential for treating humoral immunity disorders, which were mainly mediated by B lymphocytes.

Liposomes encapsulated dimethyl curcumin regulates dipeptidyl peptidase I activity, gelatinase release and cell cycle of spleen lymphocytes in-vivo to attenuate collagen induced arthritis in rats

Rheumatoid arthritis (RA) is an autoimmune disease and characterized by the excessive cell proliferation, abnormal cell cycle of lymphocytes and synovial cells. The therapeutic effects of curcumin in active RA patients were reported, but limited by its insolubility and rapid systemic elimination. Dimethyl curcumin (DiMC) is a metabolically stable analogue of curcum with anti-inflammatory property. In this study, liposomes encapsulated dimethyl curcumin (Lipo-DiMC) was prepared to improve the bioavailability and metabolic-stability; collagen induced arthritis (CIA) rat model was employed to investigate the effects of Lipo-DiMC treatments during CIA progress. Physical assessments and routine-blood-test were performed. Fresh spleen lymphocytes were isolated from normal, CIA and Lipo-DiMC-treated CIA rats; flow-cytometry for cell-cycle analysis, western-blotting for intracellular signal pathway protein expressions, gelatin-zymography for matrix-metalloproteases 2/9 (MMP-2/9) and GF-AFC for dipeptidyl-peptidase I (DPPI) activity assay.Compared with untreated CIA rats, Lipo-DiMC treatments relieved paw-swellings, suppressed the increments of immunocytes numbers and inhibited DPPI and MMP-2/9 over-activity in blood. Lipo-DiMC adjusted CIA-induced cell cycle dysfunction at G0/G1-phase and S-phase of spleen lymphocytes for CIA rats. The intracellular expression-trends of P38, P21, Bcl-2, JNK-1 and DPPI of spleen lymphocytes were observed during CIA progress with and without Lipo-DiMC administrations. Lipo-DiMC exhibited its therapeutic functions by attenuating CIA development in rats, associated with down-regulating CIA-induced lymphocytes numbers, inhibiting over-expressed of DPPI and MMP-2/9, and adjusting cell cycles. These findings provide a new insight into the mechanism of Lipo-DiMC treatment in CIA rat model and suggest that Lipo-DiMC could be considered as a potential drug for RA treatment.

Effect of Dietary Advanced Glycation end Products on Cell Cycle of Peripheral Blood Lymphocytes

Advanced glycation end products (AGEs) have been linked with autoimmune diseases. AGEs present in diet are known to be absorbed intestinally. We aimed to study effects of dietary AGEs present in repeatedly heated cooking oil on cell cycle profile of lymphocytes derived from healthy volunteers. AGEs were extracted by using Aqueous-TCAChloroform method and incubated with lymphocytes for 24 hours. All extracts showed ASF (15.42-75.18) between 355-440 nm wavelength. On incubation with AGEs, an increase in S-phase fraction of treated lymphocytes (7.67 ± 3.25) was observed in 10/10 samples analysed as compared to untreated lymphocytes (1.5 ± 1.6). The results suggest that dietary AGEs may have a role in triggering the proliferation of lymphocytes. Larger studies are required to explore role of dietary AGEs on properties of lymphocytes which may contribute to etiopathogenesis of autoimmune diseases. Such studies may provide us with a modifiable risk factor which may be altered to prevent autoimmune diseases.

Effects of Polysaccharides from Platycodon grandiflorum on Immunity-Enhancing Activity In Vitro.

The study is aimed at investigating the immunoenhancement activity of polysaccharides from Platycodon grandiflorum polysaccharides (PGPSs) in vitro. In this study, some research on lymphocyte proliferation, cell cycle, and the levels of CD4+ and CD8+ T cells were performed. Four different concentrations of PGPSs (PGPStc, PGPS60c, PGPS80c, and PGPStp) were harvested and added to peripheral blood T lymphocytes. We observed significant increases in T lymphocyte proliferation at PGPStc groups individually or synergistically with phytohemagglutinin (PHA) at most concentrations, and their lymphocyte proliferation rates were the highest. The active sites of PGPStc and PGPS60c were subsequently chosen. Then, we utilized flow cytometry to determine lymphocyte cell cycle distribution and levels of CD4+ and CD8+ T cells. At most time points, PGPStc could facilitate lymphocyte cell cycle progression from the G0/G1 phase to the S and G2/M phases and, simultaneously, increase the levels of CD4+ and CD8+ T cells. These results indicate that PGPStc enhances the immune functions, suggesting that PGPStc could be a potential immunopotentiator for further in vivo and clinical trial experiments.

Effects of Medical Diagnostic Low-dose X Rays on Human Lymphocytes: Mitochondrial Membrane Potential, Apoptosis and Cell Cycle.

Low-dose radiation is widely used across the world for the diagnosis of many diseases by means of a variety of imaging technologies. However, the harmful effects of exposure to low-dose radiation during medical examination remain controversial. The authors studied the effects of medical diagnostic low-dose x rays (i.e., 0.03, 0.05, or 0.1 mGy) after an in vitro exposure of human lymphocytes. Cells with no irradiation served as the non-irradiated control group. Three biological indicators were used to determine the effects of medical diagnostic low-dose x rays at 4, 8, 24, 48, and 72 h post-irradiation. These biological endpoints were mitochondrial membrane potential (ΔΨm), cell cycle, and apoptosis. Results indicated no changes in the ΔΨm, number of apoptotic cells, and cell cycle in lymphocytes exposed to these low doses of radiation, as compared to the corresponding non-irradiated lymphocytes at all harvest time-points. These results suggested that there were no harmful effects of the diagnostic low-dose x rays when human lymphocytes were exposed in an in vitro condition.

Immunotoxic effects of single and combined pharmaceuticals exposure on a harbor seal (Phoca vitulina) B lymphoma cell line

The potential risk of pharmaceuticals in the environment to top-predators is still largely unknown. In this study, we assessed the immunotoxic effects of ten pharmaceuticals individually and as mixtures on a harbor seal (Phoca vitulina) B lymphoma cell line. A significant reduction in lymphocyte transformation was observed following an exposure to 12,500μg/L 17α-ethinyl estradiol and 25,000μg/L naproxen. Exposure to 12,500μg/L 17α-ethinyl estradiol decreased the percentage of cell in the G0/G1 phase of the cell cycle while increasing the percentage of cells in the S phase. Carbamazepine exposure increased the amount of cells in the G2/M phase. Binary mixtures showed synergistic effects in lymphocyte transformation, cell cycle and apoptosis assays. Concentrations inducing toxic effects in the cell line were similar to those affecting fish in previous studies. A reduction of functional activities of the immune system may lead to altered host resistance to pathogens in free-ranging pinnipeds.

Effect of melatonin on the peripheral T lymphocyte cell cycle and levels of reactive oxygen species in patients with premature ovarian failure.

The objective of the present study was to observe the curative effect and mechanism of melatonin for suppression of premature ovarian failure (POF). From December 2014 to June 2015, 128 patients were consecutively diagnosed with POF in the Department of Gynaecology and Obstetrics. The patients were randomly divided into the experimental and control groups. The experimental group received melatonin tablets (1–3 mg/day), while the control group received placebo tablets. The levels of six sex hormones, percentage of T lymphocytes in the G1/M phase, and levels of reactive oxygen species (ROS) were determined at four different time-points (1 day before treatment, and at 1, 3 and 6 months after treatment) in both groups. After 6 months of treatment, the levels of luteinizing hormone and follicle-stimulating hormone were significantly decreased in the experimental group compared with the control group (P<0.05). Compared with the control group, the levels of ROS in plasma were significantly decreased in the experimental group (P<0.05). Correlation analysis showed that the levels of melatonin in peripheral blood were negatively related with the levels of ROS (rs=−0.481, P<0.05). One-year follow-up study showed that the normal excretion of ovarian hormones in the experimental group was significantly higher than that of the control group (P<0.05). In conclusion, treatment with melatonin is an effective approach to suppress POF. The potential mechanism of melatonin is inhibition of ROS production and protection of the process of normal follicle development.