Evaluation of the Immunoregulatory Capacities of Feed Microbial Materials in Porcine Intestinal Immune and Epithelial Cells

The establishment of drug-free feeding systems has been required for secure and healthy livestock production. Although functional feed materials containing microorganisms as alternatives to enhance intestinal immunity are expected to be beneficial for reducing diarrhoea caused by pathogens in weaned piglets, the effects of such materials on porcine intestinal cells have not been investigated in detail. Therefore, this work evaluated the immunoregulatory functions of microbial feed materials in porcine intestinal immune and epithelial cells. Porcine immune cells isolated from Peyer’s patches and mesenteric lymph nodes were stimulated with six different feed materials containing microorganisms, and evaluated for lymphocyte mitogenicity and cytokine inductions. In addition, porcine intestinal epithelial cells were stimulated with the materials before treatment with heat-killed enterotoxigenic Escherichia coli (ETEC), and analyzed for the proinflammatory cytokine expressions. The material containing Bifidobacterium thermophilum significantly augmented lymphocytes’ mitogenicity and also induced a high expression of IL-2, IL-6 and IFN-γ in immune cells, and inhibited ETEC-induced overexpression of IL-6 and IL-8 via regulation of Toll-like receptor signaling. These results suggest that this feed material stimulates intestinal epithelial and immune cells to exert immunoregulation, suggesting that this feed is expected to contribute to promoting the health of piglets without using antimicrobial feed materials.

Human menstrual blood-derived stem cells as immunoregulatory therapy in COVID-19:A case report and review of the literature

BACKGROUND The coronavirus disease 2019(COVID-19)caused by novel coronavirus 2019 in December 2019 has spread all around the globe and has caused a pandemic.There is still no current effective guidance on the clinical management of COVID-19.Mesenchymal stem cell therapy has been shown to be one of the therapeutic approaches to alleviate pneumonia and symptoms through their immunomodulatory effect in COVID-19 patients.CASE SUMMARY We describe the first confirmed case of COVID-19 in Hangzhou to explore the role of human menstrual blood-derived stem cells(MenSCs)in the treatment of COVID-19.Moreover,we review the immunomodulation effect including nonspecific and specific immune functions of MenSCs for the therapy of COVID-19.CONCLUSION MenSCs can be helpful to find a promising therapeutic approach for COVID-19.

Immunoregulatory Effects of Nuhuang Fuzheng Granules on Mice with Cytoxan-induced Immunosuppression

[Objectives]To study immunoregulatory effects of Nuhuang Fuzheng Granules on mice with Cytoxan-induced immunosuppression.[Methods]Kunming variety mice were randomly divided into the control group,model group,positive medicine group,and low,medium,and high dosage groups of Nuhuang Fuzheng Granules. Except the control group,the rest groups of mice were injected with Cytoxan at60 mg/kg to reproduce the immunosuppressive mice model,and were fed through stomach at 0. 5g/kg,1. 0 g/kg and 2. 0 g/kg Nuhuang Fuzheng Granules for seven days( twice a day); the positive control group was fed with Astragalus Polysacharin( 500 mg/L). The carbon clearance index,immune organ index,serum hemolysin were determined. IL-2,IFN-γ,and IL-4 levels in mouse serum were measured by ELISA. [Results] Nuhuang Fuzheng Granules could significantly incease the carbon granular expurgation index of mice with Cytoxan-induced immunosuppression,and the 2. 0 g/kg and 1. 0 g/kg dosage groups had better effect than 0. 5 g/kg dosage group( P < 0. 05); the spleen index of mice with Cytoxan-induced immunosuppression increased significantly and there was dose-effect relationship in a certain range of concentration; the thymus index and serum hemolysin HC_(50) were significantly higher than 0. 5 g/kg dosage group( P < 0. 05); IL-4 and IFN-γ in the serum was significantly higher than the model group( P < 0. 01),the increase of IL-2 was not significantly( P > 0. 05). [Conclusions] Nuhuang Fuzheng Granules can promote growth of immune organs and secretion of cytokines through improving the carbon granular expurgation ability of mice with Cytoxan-induced immunosuppression.

Structure and immunomodulatory activity of Lentinus edodes polysaccharides modified by probiotic fermentation

Plant-based fermentations provide an untapped source for novel biotechnological applications.In this study,a probiotic named Lactobacillus fermentum 21828 was introduced to ferment Lentinus edodes.Polysaccharides were extracted from fermented and non-fermented L.edodes and purified via DEAE-52 and Sephadex G-100.The components designated F-LEP-2a and NF-LEP-2a were analyzed by FT-IR,HPGPC,HPAEC,SEM,GC-MS and NMR.The results revealed that probiotic fermentation increased the molecular weight from 1.16×10^(4) Da to 1.87×10^(4) Da and altered the proportions of glucose,galactose and mannose,in which glucose increased from 45.94%to 48.16%.Methylation analysis and NMR spectra indicated that F-LEP-2a and NF-LEP-2a had similar linkage patterns.Furthermore,their immunomodulatory activities were evaluated with immunosuppressive mice.NF-LEP and F-LEP improved immune organ indices,immunoglobulin(Ig G and Ig M)and cytokines concentrations;restored the antioxidation capacity of liver;and maintained the balance of gut microbiota.F-LEP displayed better moderating effects on the spleen index,immunoglobulin,cytokines and the diversity of gut microbiota than NF-LEP(200,400 mg/kg).Our study provides an efficient and environment-friendly way for the structural modification of polysaccharides,which helps to enhance their biological activity and promote their wide application in food,medicine and other fields.

Anti-Inflammatory and Immunoregulatory Effects of Yupingfeng(玉屏风) Powder on Chronic Bronchitis Rats

Objective： To investigate the anti-inflammatory and immunoregulatory effects of Yupingfeng , Powder and its components in rats. Methods： A rat chronic bronchitis （CB） model was developed using lipopolysaccharide （LPS） combined with bacillus Calmette Guerin （BCG）. YPF, simple recipe Astragalus membranaceus （Fisch.） Bge （AM） and Astragalus membranaceus （Fisch.） Bge plus rhizome of Atractylodes macrocepha/a Koidz （AM＋RA） decoction were administered （intragastric administration, once a day for 21 days） to rats, to prevent and treat CB. Immunoregulatory and anti-inflammatory effects of YPF, AM and AM＋RA were tested by serum pharmacology in vitro on splenic lymphocytes of normal rats and alveolar macrophages of CB rats. Results： Inflammation in the pulmonary tissue and the bronchus of CB rats was significantly reduced in the YPF-treatment groups, AM and AM＋RA groups demonstrating the efficacy of YPF. Serum samples collected at different times from rats after administration of YPF, AM and AM＋RA demonstrated increased proliferation of splenic lymphocytes with area under the effect curve （AUE） of 552.6%, 336.3% and 452.0%, respectively. Treatment of alveolar macrophages with serum samples in YPF, AM or AM＋RA group inhibited interleukin-8 （IL-8） in the cell culture media, and the effect was much better in the YPF group compared with AM or AM＋RA group, with a higher maximal effect （Emax, P〈0.05） and larger AUE （P〈0.01 and P〈0.05）. Moreover, serum from rats treated with AM or AM＋RA had similar efficacy, while the efficiency was lower than that treated with YPF. Conclusion： YPF demonstrated anti-inflammatory and immunoregulatory effects in a rat model of CB, and time- dependent relationships were demonstrated in vitro.

Anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways act as potent immunoregulatory cells in vitro and vivo

Background This study was to evaluate whether anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro and prolong cardiac allograft survival after adoptive transfer KH*2/5DMethods Anergic cells were induced in vitro by the addition of anti-CD154 and anti-CD80 monoclonal antibodies (mAbs) to primary MLR (mixed lymphocyte reaction) consisting of BALB/c as responder and C3H as stimulator Anergic cells were added to a newly formed MLR in assessing the regulatory capacity and antigen specificity of anergic cells The ability of anergic cells to respond to antigen and/or exogenous recombinant mouse interleukin-2 (rmIL-2) was tested For in vivo studies, anergic cells were intravenously injected into 3 0-Gy γ-irradiated BALB/c mice immediately after heterotopic abdominal cardiac transplantation To prolong allograft survival, recipient mice injected with anergic cells received rapamycin therapy (1 mg·day -1 ·kg -1 ) KH*2/5DResults Anergic cells strongly suppressed the proliferation of naǐve BALB/c splenocytes against the original (C3H) stimulator in a dose-dependent manner, but they failed to suppress the proliferation of naǐve BALB/c splenocytes against the third-party (C57BL/6J) stimulator The anergic state was reversed by both original (C3H) stimulator and additional exogenous IL-2 In in vivo studies, untreated irradiated BALB/c mice rejected C3H cardiac allografts with a mean survival time of (8 6±1 1) days, whereas those injected with the anergic cells rejected the allografts with a mean survival time of (11 8±1 9) days, which was slightly longer than that of the untreated mice The protocol based on anergic cells injection plus rapamycin therapy could prolong allograft survival significantly [(29 6±4 4) days] Conclusions Anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro , and prolong cardiac allograft survival after adoptive transfer in the presence of rapamycin therapy This procedure might be clinically useful for prolonging allograft survival if optimal protocols are developed

Exploration of Bioactive Constituents and Immunoregulatory Mechanisms of a Hanshi-Yufei Formulation for Treating COVID-19

Objective:The objective of this study was to characterize the chemical compounds of a Hanshi-Yufei formulation(HSYF;a modified formulation of a traditional Chinese medicine used for treating COVID-19)to elucidate the mechanism of action and to evaluate potential anti-inflammatory effects of HSYF.Materials and Methods:The chemical constituents of HSYF extract were characterized using UPLC-Q-TOF/MS.Subsequently,a set of TCM network pharmacology methods was applied to identify disease-associated genes and to predict target profiles and pharmacological actions associated with the constituents of HSYF.Then,the antiviral effects of HSYF on H1N1 were assessed in RAW264.7 cells using MTT assays.Expression levels of pro-inflammatory cytokines interleukin(IL)-6 and tumor necrosis factor(TNF)-αfollowing infection of RAW264.7 cells with H1N1 were measured using an enzyme-linked immune sorbent assay(ELISA),and expression levels of inflammatory-related factors were detected using western blotting.Results:In total,165 chemical constituents(including glycosides,tannins,volatile oils,amino acids,triterpenoids,polyphenols,phenylpropanoids,sesquiterpenes,alkaloids,and flavonoids,among others)were tentatively identified in HSYF.Network pharmacology demonstrated that HSYF can regulate immunomodulatory-and anti-inflammatory-related targets of multiple pathways through its active ingredients,suggesting potential anti-COVID-19 effects.Furthermore,cell viability assays and ELISA showed that HSYF significantly inhibited H1N1 replication in RAW64.7 cells and markedly reduced expression of pro-inflammatory cytokines TNF-αand IL-6 at the proteins level.Conclusions:The results of the present study help improve our understanding of the therapeutic effects of HSYF in COVID-19 treatment from multi-level perspectives.

Structure modifications based on KRN7000 and their SARs in activating NKT cells

α-Galactosylceramides, which can be recognized by natural killer T （NKT） cells, are now attracting more and more attention due to their therapeutic potential in cancer, infection and autoimmune diseases. Advances have been achieved in discovering compounds with better activities and efforts have been made to understand the structure-activity relationships （SARs） of these NKT cell ligands. In this review, we discuss the structure modifications based on KRN7000, the principal glycolipid used in the study of NKT cell stimulation, and the SARs based on these modified structures.

Cytoprotective role of heme oxygenase-1 and heme degradation derived end products in liver injury

The activation of heme oxygenase-1(HO-1) appears to be an endogenous defensive mechanism used by cells to reduce inflammation and tissue damage in a number of injury models. HO-1, a stress-responsive enzyme that catabolizes heme into carbon monoxide(CO), biliverdin and iron, has previously been shown to protect grafts from ischemia/reperfusion and rejection.In addition, the products of the HO-catalyzed reaction, particularly CO and biliverdin/bilirubin, have been shown to exert protective effects in the liver against a number of stimuli, as in chronic hepatitis C and in transplanted liver grafts. Furthermore, the induction of HO-1 expression can protect the liver against damage caused by a number of chemical compounds. More specifically, the CO derived from HO-1-mediated heme catabolism has been shown to be involved in the regulation of inflammation; furthermore, administration of low concentrations of exogenous CO has a protective effect against inflammation. Both murine and human HO-1 deficiencies have systemic manifestations associated with iron metabolism, such as hepatic overload(with signs of a chronic hepatitis) and iron deficiency anemia(with paradoxical increased levels of ferritin).Hypoxia induces HO-1 expression in multiple rodent,bovine and monkey cell lines, but interestingly, hypoxia represses expression of the human HO-1 gene in a variety of human cell types(endothelial cells, epithelial cells, T cells). These data suggest that HO-1 and CO are promising novel therapeutic molecules for patients with inflammatory diseases. In this review, we present what is currently known regarding the role of HO-1 in liver injuries and in particular, we focus on the implications of targeted induction of HO-1 as a potential therapeutic strategy to protect the liver against chemically induced injury.

The Research of Treg Cells: Progress and Challenge

T regulatory cells (Tregs) play an important role in the induction and maintenance of immunological tolerance to self and alloantigens. Recent findings in experimental transplant models have demonstrated that Treg could control and delay allograft rejection. Induction of immune tolerance decreases the risk of acute and chronic graft rejection after solid organ transplantation and can improve transplanted organ survival. Tregs are being tested in trials as a potential therapy in cell and solid organ transplantation. However, as we know, regulatory T cells (Tregs) are crucial for peripheral tolerance and are intimately involved in cancer. The influence of Tregs on cancer progression has been demonstrated in a large number of preclinical models and confirmed in several types of malignancies. Neoplastic processes trigger an increase of Treg numbers in draining lymph nodes, spleen, blood, and tumors, leading to the suppression of anti-tumor responses. In this review, we summarize some of the critical aspects of the immunoregulatory function of Treg cells in cancer and transplantation and discuss their potential research progress and challenge.

肝癌致门静脉胆总管内瘘1例报告

肝癌致门静脉胆总管内瘘１例报告王德荣患者，男，４８岁，住院号３６７７。因右上腹不适、隐痛３年，加重１月，全身皮肤及巩膜进行性黄疽半月，于１９９４年５月１７日收住我院。病人于人院前８年患“乙型肝炎”，入院前五年诊断为“肝硬变”。入院查体：神清、消瘦、皮...

A two-pronged strategy utilizing exosomes extracted from antigenpresenting cells to combat hepatitis B

Chronic hepatitis B(CHB)is consistently challenging to conquer with numerous complications and fatalities.Despite the effectiveness of antiviral therapies in suppressing hepatitis B virus(HBV)replication,there is an urgent need for novel and more effective treatment modalities.Current strategies predominantly emphasize immune activation but still face challenges in sufficiently eliciting T cell responses.Taking into account the targeted delivery of nanoparticles to the liver and spleen via intravenous injection,we have proposed a dual-pronged therapeutic strategy based on antigen-presenting cells(APCs)-derived exosomes.Specifically,exosomes targeted to the spleen can activate specific immune responses,while those targeted to the liver can modulate or reverse the liver’s immunosuppressive microenvironment.After immunization,exosome formulations exhibit the remarkable ability to effectively activate APCs,thereby triggering the proliferation of CD8^(+)T cells.Simultaneously,they also play an immunoregulatory role by converting M2 macrophages into M1 macrophages.This two-pronged therapeutic strategy precisely addresses the issues of T cell dysfunction and immune suppression,both characteristic features of CHB patients.When combined with Aluminum(Alum)-adjuvanted vaccine,these exosome formulations not only demonstrate a high level of cellular immune response but also secrete specific antibodies comparable to those induced by Alum adjuvant.This combined approach effectively enhances both cellular and humoral immunity,offering a promising avenue for the development of therapeutic hepatitis B vaccines based on exosome formulations.

Application of immune checkpoint targets in the anti-tumor novel drugs and traditional Chinese medicine development

Immune checkpoints are the crucial regulators of immune system and play essential roles in maintaining self-tolerance,preventing autoimmune responses,and minimizing tissue damage by regulating the duration and intensity of the immune response.Furthermore,immune checkpoints are usually overexpressed in cancer cells or noninvasive cells in tumor tissues and are capable of suppressing the antitumor response.Based on substantial physiological analyses as well as preclinical and clinical studies,checkpoint molecules have been evaluated as potential therapeutic targets for the treatment of multiple types of cancers.In the last few years,extensive evidence has supported the immunoregulatory effects of traditional Chinese medicines(TCMs).The main advantage of TCMs and natural medicine is that they usually contain multiple active components,which can act on multiple targets at the same time,resulting in additive or synergistic effects.The strong immune regulation function of traditional Chinese medicine on immune checkpoints has also been of great interest.For example,Astragalus membranaceus polysaccharides can induce anti-PD-1 antibody responses in animals,and these antibodies can overcome the exhaustion of immune cells under tumor immune evasion.Furthermore,many other TCM molecules could also be novel and effective drug candidates for the treatment of cancers.Therefore,it is essential to assess the application of immune checkpoints in the development of new drugs and TCMs.In this review,we focus on research progress in the field of immune checkpoints based on three topics:(1)immune checkpoint targets and pathways,(2)development of novel immune checkpoint-based drugs,and(3)application of immune checkpoints in the development of TCMs.

Association analysis of cytokine polymorphisms and plasma level in Northern Chinese Han patients with paroxysmal nocturnal hemoglobinuria

Background While the incidence of paroxysmal nocturnal hemoglobinuria （PNH） is relatively high in Northern China, the exact mechanism of the disease remains unknown. Immunoregulatory cytokine polymorphisms can directly regulate the expression levels of cytokines, which play a crucial role in many diseases. The purpose of this study was to study cytokine gene single nucleotide polymorphisms （SNPs） and the correlated cytokine expression levels in relationship to the PNH pathogenesis.Methods Peripheral blood samples were collected from 30 PNH patients and 40 healthy donors; all of the samples were collected from the Han people of Northern China. Eight SNP loci in five cytokine genes, including tumor necrosis factor-αlpha （TNF-α）, interferon-γamma （IFN-γ）, transforming growth factor-βeta （TGF-β）, interleukin-6 （IL-6）, and IL-10, and aplastic anemia （AA） were assessed. TNF-α, TGF-β, IFN-γ, IL-6, and IL-10 were analyzed by sequence-specific primer polymerase chain reaction （PCR-SSP）. The plasma protein levels of TNF-α, TGF-β, and IFN-γ were assessed by an ELISA.Results The PNH patients had a lower frequency of the TC/GG genotype of the TGF-β gene （P 〈0.01） and a higher frequency of the C allele in the TGF-β gene （＋10） compared to the controls （P 〈0.05）. The predominant genotype of the ＋874 locus of the IFN-γ gene was TA in the PNH patients, while that in the predominant genotype was AA in the control group and was statistically significant （P 〈0.001）. The frequency of the T allele in the IFN-γ gene was dramatically higher in the PNH patients than in the controls （P 〈0.05）. The PNH patients had a reduced frequency of the GC and CC genotypes, as well as the C allele at locus –174 of the IL-6 gene compared to the controls （P 〈0.01）. In addition, the plasma concentrations of TNF-α, TGF-β, and IFN-γ were significantly higher in the PNH group compared to the control group （P 〈0.01）.Conclusions Expression levels of the TNF-α, TGF-β, and IFN-γ cytokines play an important role in PNH. The GC and CC genotypes, as well as the C allele of the IL-6 gene may protect the Han people of Northern China against PNH. Additionally, the TC/GG genotype of the TGF-β gene may be the protective allele. In contrast, the TA genotype and the T allele for the IFN-γ gene, as well as the C allele of TGF-β may be susceptible to PNH. However, SNPs in the TNF-α and IL-10 genes did not correlate with PNH development. Alternatively, the increased plasma concentrations of TNF-α, TGF-β, and IFN-γ in PNH patients may also be related to PNH development.

Programming CAR T cells to enhance anti-tumor efficacy through remodeling of the immune system

Chimeric antigen receptor(CAR)T cells have been indicated effective in treating B cell acute lymphoblastic leukemia and non-Hodgkin lymphoma and have shown encouraging results in preclinical and clinical studies.However,CAR T cells have achieved minimal success against solid malignancies because of the additional obstacles of their insufficient migration into tumors and poor amplification and persistence,in addition to antigen-negative relapse and an immunosuppressive microenvironment.Various preclinical studies are exploring strategies to overcome the above challenges.Mobilization of endogenous immune cells is also necessary for CAR T cells to obtain their optimal therapeutic effect given the importance of the innate immune responses in the elimination of malignant tumors.In this review,we focus on the recent advances in the engineering of CAR T cell therapies to restore the immune response in solid malignancies,especially with CAR T cells acting as cellular carriers to deliver immunomodulators to tumors to mobilize the endogenous immune response.We also explored the sensitizing effects of conventional treatment approaches,such as chemotherapy and radiotherapy,on CAR T cell therapy.Finally,we discuss the combination of CAR T cells with biomaterials or oncolytic viruses to enhance the anti-tumor outcomes of CAR T cell therapies in solid tumors.