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The Journal of Infection Jul 2020Distinguishing between active tuberculosis (ATB) and latent tuberculosis infection (LTBI) remains challenging.
Combination of mean spot sizes of ESAT-6 spot-forming cells and modified tuberculosis-specific antigen/phytohemagglutinin ratio of T-SPOT.TB assay in distinguishing between active tuberculosis and latent tuberculosis infection.
OBJECTIVES
Distinguishing between active tuberculosis (ATB) and latent tuberculosis infection (LTBI) remains challenging.
METHODS
The modified T-SPOT.TB assay was performed in 499 participants (243 ATB and 256 LTBI) and another 322 participants (162 ATB and 160 LTBI) who were diagnosed in Qiaokou (training) and Caidian (validation) cohort respectively.
RESULTS
The mean spot sizes (MSS) of early secreted antigenic target 6 (ESAT-6) spot-forming cells (SFC) of T-SPOT.TB assay in ATB patients was significantly higher than that in LTBI individuals. 1.0 × 10 was the optimal number of cells added to phytohaemagglutinin (PHA) well for obtaining more accurate TB-specific antigen to phytohaemagglutinin (TBAg/PHA) ratio. The area under the curve of the diagnostic model by combination of ESAT-6 SFC MSS and modified TBAg/PHA ratio in distinguishing ATB from LTBI was 0.959 in training cohort, with a sensitivity of 90.12% and a specificity of 91.02% when a cutoff value of 0.46 was used. This diagnostic model showed similar performance in the validation cohort. The area under the curve, sensitivity, and specificity were 0.962, 93.21%, and 90.00%, respectively. Further flow cytometry analysis showed that ESAT-6 stimulation induced a significantly higher mean fluorescence intensity of IFN-γ cells in lymphocytes compared with culture filtrate protein 10 (CFP-10) stimulation. In contrast, CFP-10 stimulation induced a significantly higher percentage of IFN-γ cells in lymphocytes compared with ESAT-6 stimulation.
CONCLUSIONS
The combination of the MSS of ESAT-6 SFC and the modified TBAg/PHA ratio of T-SPOT.TB assay showed great value in discriminating ATB from LTBI.
Topics: Antigens, Bacterial; Bacterial Proteins; Humans; Latent Tuberculosis; Mycobacterium tuberculosis; Phytohemagglutinins; Sensitivity and Specificity; Tuberculosis
PubMed: 32360883
DOI: 10.1016/j.jinf.2020.04.038 -
Cell Transplantation 2020Activated T lymphocytes play an important role in the pathogenesis of rheumatic diseases (RD). Mesenchymal stem cells (MSCs) possess immunoregulatory activities but such...
BACKGROUND
Activated T lymphocytes play an important role in the pathogenesis of rheumatic diseases (RD). Mesenchymal stem cells (MSCs) possess immunoregulatory activities but such functions of MSCs from bone marrow of systemic lupus erythematosus (SLE), systemic sclerosis (SSc), and ankylosing spondylitis (AS) patients are impaired. Adipose tissue-derived MSCs (ASCs) are an optional pool of therapeutically useful MSCs, but biology of these cells in RD is poorly known. This study aimed at investigating the effect of ASCs from RD patients and healthy donors (HD) on the expression of the key T-cell activation markers.
METHODS
ASCs were isolated from subcutaneous abdominal fat from SLE ( = 16), SSc ( = 18), and AS ( = 16) patients, while five human ASCs lines from HD were used as a control. Untreated and cytokine (tumor necrosis factor α + interferon γ)-treated ASCs were co-cultured with allogenic, mitogen (phytohemagglutinin)-stimulated peripheral blood mononuclear cells (PBMCs) or purified anti-CD3/CD28-activated CD4 T lymphocytes. Contacting and noncontacting ASCs-PBMCs co-cultures were performed. RD/ASCs were analyzed in co-cultures with both allogeneic and autologous PBMCs. Flow cytometry analysis was used to evaluate expression of CD25, HLA-DR, and CD69 molecules on CD4 and CD8 cells.
RESULTS
In co-cultures with allogeneic, activated CD4 T cells and PBMCs, HD/ASCs and RD/ASCs downregulated CD25 and HLA-DR, while upregulated CD69 molecules expression on both CD4 and CD8 cells with comparable potency. This modulatory effect was similar in contacting and noncontacting co-cultures. RD/ASCs exerted weaker inhibitory effect on CD25 expression on autologous than allogeneic CD4 and CD8 T cells.
CONCLUSION
RD/ASCs retain normal capability to regulate expression of activation markers on allogeneic T cells. Both HD/ASCs and RD/ASCs exert this effect independently of their activation status, mostly through the indirect pathway and soluble factors. However, autologous CD4 and CD8 T cells are partially resistant to RD/ASCs inhibition of CD25 expression, suggesting weaker control of T-cell activation .
Topics: Adult; Aged; Cells, Cultured; Female; Humans; Lymphocyte Activation; Male; Mesenchymal Stem Cells; Middle Aged; Rheumatic Diseases; T-Lymphocytes; Young Adult
PubMed: 32878464
DOI: 10.1177/0963689720945682 -
Asian Pacific Journal of Cancer... May 2022Natural killer (NK) cell receptors affect the NK cell-mediated elimination of malignant cells. In this experimental study the effect of Zoledronic acid (ZOL) was...
Reduced Expression of Natural Killer Cell-Related Activating Receptors by Peripheral Blood Mononuclear Cells from Patients with Breast Cancer and Their Improvement by Zoledronic Acid.
BACKGROUND/AIM
Natural killer (NK) cell receptors affect the NK cell-mediated elimination of malignant cells. In this experimental study the effect of Zoledronic acid (ZOL) was investigated on the expression of NK activating- (NKP46 and NKG2D) and inhibitory (KIR2DL1) receptors by Phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMCs) from breast cancer (BC) patients.
MATERIALS AND METHODS
Peripheral blood mononuclear cell-extracted RNA from thirty breast cancer women and twenty-five healthy subjects was analyzed for gene expression of NKP46, NKG2D and KIR2DL1 using real time-PCR. Then, the PBMCs from BC patients were cultured in the presence of PHA with 5 μg/ml, 10 or 20 μg/ml of ZOL for 32 hours and expression of the aforementioned receptors was determined.
RESULTS
Expression of NKP46, NKG2D and NKP46/KIR2DL1 ratio in BC women were lower than healthy group (P<0.01, P<0.04 and P<0.05, respectively). NKP46 expression was up-regulated by PHA-stimulated PBMCs treated with 10 μg/ml and 20 μg/ml of ZOL compared with PHA-stimulated cultures (P<0.01 and P<0.05, respectively). NKG2D expression remarkably increased by PHA-stimulated cultures treated with 5 μg/ml, 10 μg/ml and 20 μg/ml of ZOL compared with PHA-stimulated cultures (P<0.05 and P<0.02 and P<0.04, respectively).
CONCLUSION
Expression of NK cell-related activating receptors decreased in BC patients. ZOL can improve the expression of NK activating receptors.
Topics: Breast Neoplasms; Female; Humans; Leukocytes, Mononuclear; NK Cell Lectin-Like Receptor Subfamily K; Natural Cytotoxicity Triggering Receptor 1; Phytohemagglutinins; Receptors, KIR2DL1; Receptors, Natural Killer Cell; Zoledronic Acid
PubMed: 35633551
DOI: 10.31557/APJCP.2022.23.5.1661 -
Journal of Clinical Immunology Feb 2023Hypoparathyroidism-retardation-dysmorphism (HRD) syndrome is a disease composed of hypoparathyroidism, growth retardation, severe developmental delay, and typical...
BACKGROUND
Hypoparathyroidism-retardation-dysmorphism (HRD) syndrome is a disease composed of hypoparathyroidism, growth retardation, severe developmental delay, and typical dysmorphic features caused by the tubulin-specific chaperone E gene variant. Many patients succumb in infancy to HRD due to overwhelming infections mainly caused by Pneumococcus spp. Knowledge related to the immune system in these patients is scarce.
PURPOSE
To define the immune phenotype of a cohort of HRD patients including their cellular, humoral, and neutrophil functions.
METHODS
The study included HRD patients followed at Soroka University Medical Center. Clinical and immunological data were obtained, including immunoglobulin concentrations, specific antibody titers, lymphocyte subpopulations, lymphocyte proliferation, and neutrophil functions.
RESULTS
Nine patients (5 females and 4 males) were enrolled, aged 6 months to 15 years. All received amoxicillin prophylaxis as part of a routine established previously. Three patients had bacteremia with Klebsiella, Shigella spp., and Candida. Three patients had confirmed coronavirus disease 19 (COVID-19), and two of them died from this infection. All patients had normal blood counts. Patients showed high total IgA and IgE levels, low anti-pneumococcal antibodies in spite of a routine vaccination schedule, and reduced frequency of naive B cells with increased frequency of CD21lowCD27- B cells. All patients had abnormal T-cell population distributions, including reduced terminally differentiated effector memory CD8, inverted CD4/CD8 ratios, and impaired phytohemagglutinin (PHA)-induced lymphocyte proliferation. Neutrophil superoxide production and chemotaxis were normal in all patients tested.
CONCLUSION
HRD is a combined immunodeficiency disease with syndromic features, manifesting in severe invasive bacterial and viral infections.
Topics: Male; Female; Humans; Tubulin; COVID-19; Growth Disorders; Hypoparathyroidism
PubMed: 36258138
DOI: 10.1007/s10875-022-01380-9 -
Frontiers in Pediatrics 2022Patients with T cell deficiency <10% of normal proliferation are indicated to receive immune reconstruction by hematopoietic stem cell transplantation (HSCT). This study...
BACKGROUND
Patients with T cell deficiency <10% of normal proliferation are indicated to receive immune reconstruction by hematopoietic stem cell transplantation (HSCT). This study aimed to investigate whether non-radioactive assays can be used to quantitatively detect the lymphocyte proliferation <10% of normal as radioactive [H]-thymidine."
METHODS
Radioactive [H]-thymidine, non-radioactive carboxyfluorescein diacetate succinimidyl ester (CFSE), and Ki-67 protein expressions were used to measure the lymphocyte proliferation as calculated using the stimulation index (SI), subtraction percentage, and proliferation index (FlowJo software). Normal references were established for comparison in the absence of parallel healthy controls.
RESULTS
Normal ranges of mitogen-stimulated lymphocyte proliferation were established as a SI of 15-267 (CSFE 47-92%, Ki-67 42-79%) with phytohemagglutinin (PHA) 5 μg/ml stimulation; 19-139 (CFSE 62-83%, 45-74% Ki-67) with concanavalin-A (ConA) 5 μg/ml stimulation; 7-53 (CFSE 6-23%, Ki-67 10-24%) with pokeweed mitogen (PWM) 0.1 ug/ml stimulation; 3-28 (CFSE 4-10%, Ki-67 5-14%) with candida 10 ug/ml stimulation; and 2-27 (CFSE 6-41%, Ki-67 6-30%) with bacille Calmette-Guerin (BCG) 0.02 ng/ml stimulation. The normalized CFSE-proliferation index was between 2.1 and 3.0. Although there was no significant correlation between these three assays in the healthy controls, the SI value for <10% [H]-thymidine proliferation in those with T cell deficiency was compatible with CFSE- and Ki-67-stained lymphocyte percentages, and validated in patients with , and mutations. When calculating [H]-thymidine <10% of normal lymphocyte proliferation, the threshold of parallel controls was more reliable than previously established normal references.
CONCLUSION
The large quantitative value of radioactive [H]-thymidine was more easily recognizable than that for non-radioactive CFSE and Ki-67. Even though the correlation was not significant, those identified to have <10% of normal proliferation by [H]-thymidine could be consistently detected by CFSE and Ki-67, and consequently indicated for HSCT.
PubMed: 35547552
DOI: 10.3389/fped.2022.638549 -
Frontiers in Immunology 2021The small molecule cyclotriazadisulfonamide (CADA) down-modulates the human CD4 receptor, an important factor in T cell activation. Here, we addressed the...
Small Molecule Cyclotriazadisulfonamide Abrogates the Upregulation of the Human Receptors CD4 and 4-1BB and Suppresses In Vitro Activation and Proliferation of T Lymphocytes.
The small molecule cyclotriazadisulfonamide (CADA) down-modulates the human CD4 receptor, an important factor in T cell activation. Here, we addressed the immunosuppressive potential of CADA using different activation models. CADA inhibited lymphocyte proliferation with low cellular toxicity in a mixed lymphocyte reaction, and when human PBMCs were stimulated with CD3/CD28 beads, phytohemagglutinin or anti-CD3 antibodies. The immunosuppressive effect of CADA involved both CD4 and CD8 T cells but was, surprisingly, most prominent in the CD8 T cell subpopulation where it inhibited cell-mediated lympholysis. Immunosuppression by CADA was characterized by suppressed secretion of various cytokines, and reduced CD25, phosphoSTAT5 and CTPS-1 levels. We discovered a direct down-modulatory effect of CADA on 4-1BB (CD137) expression, a survival factor for activated CD8 T cells. More specifically, CADA blocked 4‑1BB protein biosynthesis by inhibition of its co-translational translocation into the ER in a signal peptide-dependent way. Taken together, this study demonstrates that CADA, as potent down-modulator of human CD4 and 4‑1BB receptor, has promising immunomodulatory characteristics. This would open up new avenues toward chemotherapeutics that act as selective protein down-modulators to treat various human immunological disorders.
Topics: CD4 Antigens; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Proliferation; Cells, Cultured; Cytokines; Endoplasmic Reticulum; HEK293 Cells; Humans; Jurkat Cells; Leukocytes, Mononuclear; Lymphocyte Activation; Protein Transport; STAT5 Transcription Factor; Small Molecule Libraries; Sulfonamides; Tumor Necrosis Factor Receptor Superfamily, Member 9; Up-Regulation
PubMed: 33968048
DOI: 10.3389/fimmu.2021.650731 -
Scientific Reports Apr 2021This experiment aimed to investigate the effect of dietary Nigella sativa on the cell-mediated immune response. Eighteen male Wistar rats were divided equally into a...
This experiment aimed to investigate the effect of dietary Nigella sativa on the cell-mediated immune response. Eighteen male Wistar rats were divided equally into a control group and treated groups that received black seeds at rates of 30 and 50 g/kg in the diet (Sa30 and Sa50 groups, respectively, for 30 days. The weight gain, feed intake, feed conversion ratio (FCR), and cell-mediated immune response were monitored after the injection of 0.1 mL of 10% phytohemagglutinin (PHA). The intumesce index, serum total antioxidant capacity (TAC), catalase (CAT), interleukin-12 (IL-12), gamma interferon (γ-IF) and tumor necrosis factor alpha (TNF-α) were determined. Histopathological examination and an immunohistochemistry analysis of splenic caspase-3 and CD8 were performed. Nigella sativa significantly improved the weight gain and FCR. Intumesce index of Sa50 group was significantly increased. Nigella sativa significantly increased TAC, CAT, IL-12, γ-IF and TNF-α. A histological examination of PHA-stimulated foot pads showed increased leukocyte infiltration and edema in a dose-dependent pattern. Splenic caspase-3 and CD8 showed significant decreases and increases, respectively, in the Sa30 and Sa50 groups. The results indicate that Nigella sativa seeds exhibit immunostimulatory function through their antioxidant potential, induction of cytokine production, promotion of CD8 expression and reduction of splenic apoptosis.
Topics: Adjuvants, Immunologic; Animals; Antioxidants; Cytokines; Diet; Dietary Supplements; Immunity; Male; Nigella sativa; Plant Extracts; Rats; Rats, Wistar; Seeds; Spleen
PubMed: 33824353
DOI: 10.1038/s41598-021-86721-1 -
Frontiers in Immunology 2022Thermal injury induces concurrent inflammatory and immune dysfunction, which is associated with adverse clinical outcomes. However, these effects in the pediatric... (Observational Study)
Observational Study
Thermal injury induces concurrent inflammatory and immune dysfunction, which is associated with adverse clinical outcomes. However, these effects in the pediatric population are less studied and there is no standard method to identify those at risk for developing infections. Our goal was to better understand immune dysfunction and identify soluble protein markers following pediatric thermal injury. Further we wanted to determine which early inflammatory, soluble, or immune function markers are most predictive of the development of nosocomial infections (NI) after burn injury. We performed a prospective observational study at a single American Burn Association-verified Pediatric Burn Center. A total of 94 pediatric burn subjects were enrolled and twenty-three of those subjects developed a NI with a median time to diagnosis of 8 days. Whole blood samples, collected within the first 72 hours after injury, were used to compare various markers of inflammation, immune function, and soluble proteins between those who recovered without developing an infection and those who developed a NI after burn injury. Within the first three days of burn injury, innate and adaptive immune function markers (ex vivo lipopolysaccharide-induced tumor necrosis factor alpha production capacity, and ex vivo phytohemagglutinin-induced interleukin-10 production capacity, respectively) were decreased for those subjects who developed a subsequent NI. Further analysis of soluble protein targets associated with these pathways displayed significant increases in soluble CD27, BTLA, and TIM-3 for those who developed a NI. Our findings indicate that suppression of both the innate and adaptive immune function occurs concurrently within the first 72 hours following pediatric thermal injury. At the same time, subjects who developed NI have increased soluble protein biomarkers. Soluble CD27, BTLA, and TIM-3 were highly predictive of the development of subsequent infectious complications. This study identifies early soluble protein makers that are predictive of infection in pediatric burn subjects. These findings should inform future immunomodulatory therapeutic studies.
Topics: Biomarkers; CD27 Ligand; Child; Cross Infection; Hepatitis A Virus Cellular Receptor 2; Humans; Lipopolysaccharides; Phytohemagglutinins; Prospective Studies; Receptors, Immunologic
PubMed: 35958579
DOI: 10.3389/fimmu.2022.940835 -
Journal of Ethnopharmacology Jan 2020The fruit of Astrocaryum aculeatum G.Mey. (tucumã) is highly consumed by riverside communities in the Amazonian region. These communities have recently been shown to...
ETHNOPHARMACOLOGICAL RELEVANCE
The fruit of Astrocaryum aculeatum G.Mey. (tucumã) is highly consumed by riverside communities in the Amazonian region. These communities have recently been shown to have increased longevity and reduced prevalence of age-related morbidity. Tucumã, which is locally used in their diet and traditional medicine may contribute to these features.
AIM OF THE STUDY
To investigate the anti-inflammatory and antioxidant properties of A. aculeatum extract against phytohemagglutinin-induced inflammation in cell cultures.
MATERIALS AND METHODS
Cell viability and cytotoxicity assays, gene expression of interleukins IL-1β, IL-6, IL-10, levels of reactive oxygen species (ROS), nitric oxide (NO) and thiols were employed, as well as the activities of antioxidant enzymes in RAW 264.7 cells stimulated with phytohemagglutinin to mimic inflammation.
RESULTS
The extract of A. aculeatum fruit inhibited macrophage proliferation (P < 0.05), arrested the cell cycle in G0/G1 phase (P < 0.001), increased antioxidant defenses (P < 0.01), reduced oxidative stress (P < 0.01), and modulated genes related to the inflammatory response (P < 0.001).
CONCLUSION
Our results demonstrate that A. aculeatum fruit has anti-inflammatory and antioxidant capacities. These beneficial effects of tucumã on cells are also likely to be seen in vivo, thereby suggesting that its extract is a suitable therapeutic adjuvant in the prevention or treatment of inflammatory diseases.
Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Arecaceae; Cell Survival; Drug Evaluation, Preclinical; Ethnopharmacology; Fruit; Inflammation; Medicine, Traditional; Mice; Oxidation-Reduction; Oxidative Stress; Phytohemagglutinins; Plant Extracts; Plants, Edible; RAW 264.7 Cells; South America
PubMed: 31589969
DOI: 10.1016/j.jep.2019.112274 -
Aging Nov 2019Aging is a universal and complex process that affects all tissues and cells types, including immune cells, in a process known as immunosenescence. However, many aspects...
Aging is a universal and complex process that affects all tissues and cells types, including immune cells, in a process known as immunosenescence. However, many aspects of immunosenescence are not completely understood, as the characteristics of the immune cells of nonagenarians and centenarians or the features and implications of extracellular vesicles (EVs). In this study, we analyzed blood samples from 51 individuals aged 20-49 and 70-104 years. We found that senescent CD8 cells accumulate with age, while there is a partial reduction of senescent CD4 cells in nonagenarians and centenarians. Moreover, plasma EVs carry T cell specific markers, but no accumulation of "senescent-like EVs" was found within any of analyzed age groups. Our functional studies of cocultures of peripheral blood mononuclear cells and EVs showed that EVs enhance T cell viability and, under phytohemagglutinin stimulation, they influence cytokine secretion and cell activation in an age-dependent manner. These results underline the importance of EVs on the immune system functioning, and open new perspectives to further study their implication in human aging.
Topics: Adult; Aged; Aged, 80 and over; Extracellular Vesicles; Female; Humans; Immunosenescence; Lymphocyte Activation; Male; Middle Aged; T-Lymphocytes
PubMed: 31785146
DOI: 10.18632/aging.102517