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Animal Biotechnology Nov 2024The current study was performed to determine the effect of dietary vitamin E, sesamin and thymoquinone bioactive lignans derived from sesame and black seed on...
Evaluation the effect of dietary vitamin E, sesamin and thymoquinone bioactive compounds on immunological response, intestinal traits and MUC-2 gene expression in broiler Japanese quails ().
The current study was performed to determine the effect of dietary vitamin E, sesamin and thymoquinone bioactive lignans derived from sesame and black seed on immunological response, intestinal traits and Mucin gene expression in broiler quails. Three hundred and fifty (one days-old) quails were allotted to seven dietary treatments with five replicates as an experimental randomized design study. Treatments were basal diet as a control, control +100 and +200 mg of vitamin E, sesamin and thymoquinone per each kg of diet respectively. At 35 d of age, two quails from each pen were chosen, weighted, slaughtered, eviscerated and lymphoid organ relative weights were measured. Anti-body titers against Newcastle disease (ND), Sheep red blood cell (SRBC), and infectious bronchitis virus (IBV) and Avian influenza (AI) vaccination were determined. The serum activities of alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and serum antioxidant activates such as superoxide dismutase (SOD),glutathione peroxidase(GPX), catalase (CAT) and total antioxidant capacity (TAC) were examined. The cell mediated immunity by dinitrochlorobenzene (DNCB) and phytohemagglutinin (PHA) challenges were assessed. The microflora populations of ileum, morphological traits of jejunum and mucin gene expression were analyzed. Data showed that the lymphoid organ (thymus, spleen and Bursa) relative weights and antibody titer against HI, AI, SRBC and IB vaccination were increased compared to the control ( ≤ 0.05). Serum activities of ALP, ALT and AST were decreased under influences of dietary treatments ( ≤ 0.05). The serum antioxidant activates of GPX,SOD,CAT and TAC were increased and Increasing in mean skin thickness after DNCB challenge and decrease wing web swelling response to PHA mitojen injection were observed ( ≤ 0.05). and colonies were decrease and colonies increased instead ( ≤ 0.05). The villus height and surface, crypt depth and goblet cells density were increased compared to the control ( ≤ 0.05). The expression of MUC gene increased under influnces of vitamin E, sesamin and thymoquinone supplemented diets ( ≤ 0.05).
Topics: Animals; Sheep; Coturnix; Vitamin E; Antioxidants; Dinitrochlorobenzene; Chickens; Diet; Dietary Supplements; Lignans; Superoxide Dismutase; Gene Expression; Mucins; Animal Feed; Dioxoles; Benzoquinones
PubMed: 37729462
DOI: 10.1080/10495398.2023.2259437 -
Fish & Shellfish Immunology Jul 2024The interleukin-12 (IL-12) family is a class of heterodimeric cytokines that play crucial roles in pro-inflammatory and pro-stimulatory responses. Although some IL-12...
The interleukin-12 (IL-12) family is a class of heterodimeric cytokines that play crucial roles in pro-inflammatory and pro-stimulatory responses. Although some IL-12 and IL-23 paralogues have been found in fish, their functional activity in fish remains poorly understood. In this study, Pf_IL-12p35a/b, Pf_IL-23p19 and Pf_IL-12p40a/b/c genes were cloned from yellow catfish (Pelteobagrus fulvidraco), four α-helices were found in Pf_IL-12p35a/b and Pf_IL-23p19. The transcripts of these six genes were relatively high in mucus and immune tissues of healthy individuals, and in gill leukocytes. Following Edwardsiella ictaluri infection, Pf_IL-12p35a/b and Pf_IL-23p19 mRNAs were induced in brain and kidney (or head kidney), Pf_IL-12p40a mRNA was induced in gill, and Pf_IL-12p40b/c mRNAs were induced in brain and liver (or skin). The mRNA expression of these genes in PBLs was induced by phytohaemagglutinin (PHA) and polyinosinic-polycytidylic acid (poly I:C), while lipopolysaccharides (LPS) induced the mRNA expression of Pf_IL-12p35a and Pf_IL-12p40b/c in PBLs. After stimulation with recombinant (r) Pf_IL-12 and rPf_IL-23 subunit proteins, either alone or in combination, mRNA expression patterns of genes related to T helper cell development exhibited distinct differences. The results suggest that Pf_IL-12 and Pf_IL-23 subunits may play important roles in regulating immune responses to pathogens and T helper cell development.
Topics: Animals; Catfishes; Fish Proteins; Enterobacteriaceae Infections; Fish Diseases; Gene Expression Regulation; Interleukin-12 Subunit p40; Gene Expression Profiling; Immunity, Innate; Edwardsiella ictaluri; Interleukin-12 Subunit p35; Phylogeny; Amino Acid Sequence; Sequence Alignment; Interleukin-23 Subunit p19; Poly I-C
PubMed: 38750705
DOI: 10.1016/j.fsi.2024.109623 -
Frontiers in Immunology 2023Considering the similarities between swine and humans, it is a logical consequence to use swine as a translational model in research and drug development, including...
Considering the similarities between swine and humans, it is a logical consequence to use swine as a translational model in research and drug development, including non-clinical safety. Here, we compared the reactivity of peripheral blood mononuclear cells (PBMCs) from humans and minipigs under the influence of different compounds . We conducted a flow cytometry-based proliferation assay that focused on the T-cell response to three different stimuli: concanavalin A (ConA), phytohemagglutinin-L (PHA-L), and staphylococcal Enterotoxin B (SEB). Furthermore, four approved immunosuppressive drugs-abatacept, belatacept, rapamycin, and tofacitinib-which are used for the treatment of rheumatoid arthritis or rejection in transplant recipients, were combined with the different stimuli. This allowed us to study the effect of suppressive drugs in comparison with the different stimuli in both species. We examined proliferating T cells (CD3) and investigated the presence of TCR-αβ and TCR-γδ T cells. Differences in the response of T cells of the two species under these various conditions were evident. CD4 T cells were more activated within humans, whereas CD8 T cells were generally more abundant in swine. The effectiveness of the used humanized antibodies is most likely related to the conserved structure of CTLA-4 as abatacept induced a much stronger reduction in swine compared with belatacept. The reduction of proliferation of rapamycin and tofacitinib was highly dependent on the used stimuli. We further investigated the effect of the immunosuppressive compounds on antigen-specific restimulation of pigs immunized against porcine circovirus 2 (PCV2). Treatment with all four compounds resulted in a clear reduction of the proliferative response, with rapamycin showing the strongest effect. In conclusion, our findings indicate that the effectiveness of suppressive compounds is highly dependent on the stimuli used and must be carefully selected to ensure accurate results. The results highlight the importance of considering the response of T cells in different species when evaluating the potential of an immunomodulatory drug.
Topics: Humans; Swine; Animals; Swine, Miniature; Abatacept; Leukocytes, Mononuclear; CD8-Positive T-Lymphocytes; Immunosuppressive Agents; Sirolimus; Receptors, Antigen, T-Cell
PubMed: 38264655
DOI: 10.3389/fimmu.2023.1327776 -
Proteome Science Jan 2024Tepary bean (Phaseolus acutifolius A. Gray) is one of the five species domesticated from the genus Phaseolus with genetic resistance to biotic and abiotic stress. To...
BACKGROUND
Tepary bean (Phaseolus acutifolius A. Gray) is one of the five species domesticated from the genus Phaseolus with genetic resistance to biotic and abiotic stress. To understand the mechanisms underlying drought responses in seed storage proteins germinated on water and polyethylene glycol (PEG-6000) at -0.49 MPa, we used a proteomics approach to identify potential molecular target proteins associated with the low water potential stress response.
METHODS
Storage proteins from cotyledons of Tepary bean seeds germinated at 24, 48 and 72 h on water and PEG-6000 at -0.49 MPa were analyzed by one-dimensional electrophoresis (DE) with 2-DE analysis and shotgun mass spectrometry. Using computational database searching and bioinformatics analyses, we performed Gene Ontology (GO) and protein interactome (functional protein association network) String analyses.
RESULTS
Comparative analysis showed that the effect of PEG-6000 on root growth was parallel to that on germination. Based on the SDS‒PAGE protein banding patterns and 2-DE analysis, ten differentially abundant seed storage proteins showed changes in storage proteins, principally in the phaseolin and lectin fractions. We found many proteins that are recognized as drought stress-responsive proteins, and several of them are predicted to be intrinsically related to abiotic stress. The shotgun analysis searched against UniProt's legume database, and Gene Ontology (GO) analysis indicated that most of the seed proteins were cytosolic, with catalytic activity and associated with carbohydrate metabolism. The protein‒protein interaction networks from functional enrichment analysis showed that phytohemagglutinin interacts with proteins associated with the degradation of storage proteins in the cotyledons of common bean during germination.
CONCLUSION
These findings suggest that Tepary bean seed proteins provide valuable information with the potential to be used in genetic improvement and are part of the drought stress response, making our approach a potentially useful strategy for discovering novel drought-responsive proteins in other plant models.
PubMed: 38195472
DOI: 10.1186/s12953-023-00225-6 -
International Journal of Pharmaceutics Jan 2024Immunological safety of nanofibers remains poorly reported within the scientific literature and lacks specific in vitro testing models distinct from those used to test...
Immunological safety of nanofibers remains poorly reported within the scientific literature and lacks specific in vitro testing models distinct from those used to test nanoparticles. To address the challenges of currently used conventional setups being described in the literature, we developed a novel in vitro model for nanofiber mats immunogenicity testing, which enables standardization of tested surface area, excludes nanofiber mat edges, and ensures stable contacts of cells with nanofibers during the experiment. The effect of nanofibers was assessed on peripheral blood mononuclear cells (PBMCs) by measuring their metabolic activity using MTS cell proliferation assay, where key performance parameters, i.e. cell number, phytohemagglutinin-L (PHA-L) concentration, incubation time and cell lysis were optimized. Repeatability of results obtained with non-activated and PHA-L-activated PBMCs in contact with differently thick polycaprolactone nanofiber mats was compared using both models. Our model provided more reproducible results with lower variability, exhibiting its higher reliability and accuracy than the conventional one. Furthermore, results showed the presence of thicker mats resulted in reduced metabolic activity and PBMC proliferation without any observed cytotoxicity, providing additional insights into their non-immunogenic characteristics. The developed model enables more accurate biological assessment that can support new guidelines for in vitro nanofiber testing and formulation.
Topics: Nanofibers; Leukocytes, Mononuclear; Reproducibility of Results; Nanoparticles
PubMed: 38086494
DOI: 10.1016/j.ijpharm.2023.123696 -
Heliyon Feb 2024The seaweeds are in focus for their immunity and gut health-stimulating potentials in humans and farm animals, but their potential as a gut health-promoting agent and...
The seaweeds are in focus for their immunity and gut health-stimulating potentials in humans and farm animals, but their potential as a gut health-promoting agent and performance booster to replace antibiotic growth promoters (AGP) in broiler chicken-feed remains to be evaluated. feeding experiments were conducted on commercial broiler chickens (1-42 days post-hatch) to evaluate dried aqueous exact of red seaweed (referred to as PBD 5). Each of the three test diets (basal diet with three dosing regimens of PBD5, 0.25 g kg for 0-6 weeks, 0.25 g kg for 0-4 weeks or 1.0 g kg for 0-2 weeks), along with an AGP supplemented diet (Virginiamycin (V), 20 ppm in basal diet), and a control diet was fed to 13 pen replicates of five chicks in each. PBD5 at 1.0 g kg diet for 0-2 weeks improved (P < 0.05) cumulative feed efficiency (4.65 % improvement at 28 d, and 3.74 % at 35 d) than the control and comparable to the V group and the trend in improvement persisted up to 42 d. The group fed with PBD5 @ 1.0 g kg for 0-2 weeks had significantly (P < 0.05) higher serum IgG level, glutathione peroxidase levels, fat digestibility, and expression of occludin and avian beta-defensin 4 gene in the gut and a trend of increased expression of growth hormone receptor gene in the liver as compared to the control with no significant effect on body weight, phytohemagglutinin response or haemagglutination inhibition titer. At d 25 of age, fecal count was significantly (P < 0.01) lower in the seaweed extract groups and the V group as compared to the control. It can be concluded that dried aqueous extract of at 1 g kg diet for 0-2 weeks can be used as an alternative to antibiotic growth promoter in broiler chickens to improve feed efficiency and reduce gut pathogen load, and the improved performance was associated with increased expression of gut immunity and growth hormone receptor genes.
PubMed: 38333794
DOI: 10.1016/j.heliyon.2024.e25219 -
Journal of the Chinese Medical... Oct 2023Progesterone-stimulated rapid suppression of phytohemagglutinin (PHA)-activated sustained membrane Ca 2+ influx is revealed by Mn 2+ quenching fura-2 fluorescence. Ca 2+...
BACKGROUND
Progesterone-stimulated rapid suppression of phytohemagglutinin (PHA)-activated sustained membrane Ca 2+ influx is revealed by Mn 2+ quenching fura-2 fluorescence. Ca 2+ influx suppression results in immunosuppression of T-cell proliferation. Downregulation of protein kinase C (PKC) activity by phorbol 12-myristate 13-acetate (PMA) enhances the PHA-activated increase in sustained intracellular Ca 2+ concentration ([Ca 2+ ] i ) via Ca 2+ influx in T cells. Conventional PKC (cPKC) inhibitors also enhance the [Ca 2+ ] i increase in resting T cells caused by progesterone. This study explores whether cPKC activation by progesterone results in suppression of Ca 2+ influx in resting T cells.
METHODS
Progesterone, its analogs (R5020/Org OD 02-0), and plasma membrane-impermeable progesterone-bovine serum albumin conjugate were used to stimulate human resting T cells. Inhibitors and PKC downregulation by PMA were used to investigate whether cPKC affects Ca 2+ influx.
RESULTS
Progesterone and analogs dose-dependently suppressed Ca 2+ influx in T cells. One cPKC inhibitor, Ro318220, attenuated Ca 2+ influx suppression, and enhanced the increase in [Ca 2+ ] i caused by progesterone and analogs. U73122 did not affect Ca 2+ influx suppression but did decrease the [Ca 2+ ] i increase. Ca 2+ influx suppression was not attenuated by the cPKCα/βI isoform-selective inhibitor, Go6976, nevertheless, a cPKCβI/βII isoform-selective inhibitor, LY333531 did. Ca 2+ influx suppression was attenuated by the cPKCβII-specific inhibitor CGP53353. After PKC downregulated by PMA, Ca 2+ influx suppression by progesterone and analogs was almost abolished in parallel with a massive reduction in cPKCβII expression. This suggests cPKCβII activation by progesterone and analogs mediate Ca 2+ influx suppression in resting T cells.
CONCLUSION
Nongenomic membrane activation of cPKCβII by progesterone causes immunosuppression via negative regulation of Ca 2+ influx into human resting T cells. This prevents resting T-cell activation and proliferation, which protects the fetus from maternal immune attack while decreasing maternal autoimmune disease flare-ups during pregnancy. Thus, cPKCβII modulators might provide a new therapeutic approach to balancing T-cell tolerance and immunity.
PubMed: 37496123
DOI: 10.1097/JCMA.0000000000000970 -
Cureus Dec 2023In the last couple of decades, much progress has been made in studying bacteria living in humans. However, there is much more to learn about bacteria immune cell...
In the last couple of decades, much progress has been made in studying bacteria living in humans. However, there is much more to learn about bacteria immune cell interactions. Here, we show that anaerobic bacteria do not grow when cultured overnight with human cells under atmospheric air. Air contains about 18% oxygen, which inhibits the growth of these bacteria while supporting the cultivation of human cells. The bacteria cultured with human peripheral blood mononuclear cells (PBMCs) inflamed with phytohemagglutinin (PHA) greatly increased the production of proinflammatory cytokines like tumor necrosis factor-alpha (TNFα) while inhibiting the production of monocyte chemoattractant protein-1 (MCP-1), an important chemokine.
PubMed: 38222203
DOI: 10.7759/cureus.50586 -
Bioengineering (Basel, Switzerland) Aug 2023Mesenchymal stromal cell (MSC)-derived products, such as trophic factors (MTFs), have anti-inflammatory properties that make them attractive for cell-free treatment....
Mesenchymal stromal cell (MSC)-derived products, such as trophic factors (MTFs), have anti-inflammatory properties that make them attractive for cell-free treatment. Three-dimensional (3D) culture can enhance these properties, and large-scale expansion using a bioreactor can reduce manufacturing costs. Three lots of MTFs were obtained from umbilical cord MSCs produced by either monolayer culture (Monol MTF) or using a 3D microcarrier in a spinner flask dynamic system (Bioreactor MTF). The resulting MTFs were tested and compared using anti-inflammatory potency assays in two different systems: (1) a phytohemagglutinin-activated peripheral blood mononuclear cell (PBMNC) system and (2) a lipopolysaccharide (LPS)-activated macrophage system. Cytokine expression by macrophages was measured via RT-PCR. The production costs of hypothetical units of anti-inflammatory effects were calculated using the percentage of TNF-α inhibition by MTF exposure. Bioreactor MTFs had a higher inhibitory effect on TNF ( < 0.01) than monolayer MTFs ( < 0.05). The anti-inflammatory effect of Bioreactor MTFs on IL-1β, TNF-α, IL-8, IL-6, and MIP-1 was significantly higher than that of monolayer MTFs. The production cost of 1% inhibition of TNF-α was 11-40% higher using monolayer culture compared to bioreactor-derived MTFs. A 3D dynamic culture was, therefore, able to produce high-quality MTFs, with robust anti-inflammatory properties, more efficiently than monolayer static systems.
PubMed: 37627815
DOI: 10.3390/bioengineering10080930 -
Journal of Natural Medicines Mar 2024Chemotherapy is still a prevalent strategy for clinical lung cancer treatment. However, the inevitable emerged drug resistance has become a great hurdle to therapeutic...
Chemotherapy is still a prevalent strategy for clinical lung cancer treatment. However, the inevitable emerged drug resistance has become a great hurdle to therapeutic effect. Studies have demonstrated that the primary cause of drug resistance is a decrease in the chemotherapeutic medicine concentration. Several lectins have been confirmed to be effective as chemotherapy adjuvants, enhancing the anti-tumor effects of chemotherapy drugs. Here, we combined phytohemagglutinin (PHA), which has been reported possess anti-tumor effects, with chemotherapy drugs Cisplatin (DDP) and Adriamycin (ADM) on lung cancer cells to detect the sensitivities of PHA as a chemotherapy adjuvant. Our results demonstrated that the PHA significantly enhanced the sensitivity of lung cancer cells to DDP and ADM, and Western blot showed that PHA combined with DDP or ADM enhance cytotoxic effects by inhibiting autophagy and promoting apoptosis. More importantly, we found PHA enhanced the chemotherapeutic drugs cytotoxicity by changing the cell membrane to increase the intracellular chemotherapeutic drugs concentration. Besides, the combination of PHA and ADM increased the ADM concentration in the multidrug-resistant strain A549-R cells and achieved the drug sensitization effect. Our results suggest that PHA combined with chemotherapy can be applied in the treatment of lung cancer cells and lung cancer multidrug-resistant strains, and provide a novel strategy for clinical tumor chemotherapy and a new idea to solve the problem of drug resistance in clinical lung cancer.
Topics: Humans; Lung Neoplasms; Phytohemagglutinins; Phaseolus; Cell Membrane Permeability; Drug Resistance, Neoplasm; Cell Line, Tumor; Antineoplastic Agents; Doxorubicin; Apoptosis; Cell Proliferation
PubMed: 38265611
DOI: 10.1007/s11418-023-01772-0