Similar Immunomodulatory Effects Research Articles

Immunomodulatory effects of antiangiogenic tyrosine kinase inhibitors in renal cell carcinoma models: Impact on following anti-PD-1 treatments

The approval of immune checkpoint inhibitors (ICIs) has revolutionized the management of metastatic renal cell carcinoma (RCC), introducing several ICI-based combinations as the new standard of care for affected patients. Nonetheless, monotherapy with antiangiogenic tyrosine kinase inhibitors (TKIs), such as pazopanib or sunitinib, still represents a first-line treatment option for selected patients belonging to the favorable risk group according to the International mRCC Database Consortium (IMDC) model. After TKI monotherapy, the main second-line option is represented by ICI monotherapy with the anti-Programmed Death Receptor 1(PD-1) nivolumab.To date, the expected clinical outcomes are similar with pazopanib or sunitinib and there is no clear indication for selecting one TKI over the other. Moreover, their impact on subsequent ICI treatment outcomes is not well defined, yet.Based on these premises, we investigated the immunomodulatory activity of these drugs in vitro and in vivo.Both TKIs induced Programmed Cell Death Ligand-1 (PD-L1) expression and soluble PD-L1 release in RCC cells, and hampered T cell activation, reducing cytokine production and the proportion of activated T cells. Nevertheless, in a syngeneic co-culture system with peripheral blood mononuclear cells (PBMCs) and tumor cells, incubation with anti-PD-1 antibody following TKIs treatment significantly restored T cell function, potentiating the cytotoxic effects against tumor cells. Pazopanib and sunitinib followed by anti-PD-1 antibody produced a comparable inhibition of tumor growth in a RCC syngeneic mouse model. Our findings suggest that pazopanib and sunitinib, showing similar immunomodulatory effects, may have a comparable impact on the subsequent effectiveness of PD-1/PD-L1 blockade.

Mesaconate is synthesized from itaconate and exerts immunomodulatory effects in macrophages.

Since its discovery in inflammatory macrophages, itaconate has attracted much attention due to its antimicrobial and immunomodulatory activity1-3. However, instead of investigating itaconate itself, most studies used derivatized forms of itaconate and thus the role of non-derivatized itaconate needs to be scrutinized. Mesaconate, a metabolite structurally very close to itaconate, has never been implicated in mammalian cells. Here we show that mesaconate is synthesized in inflammatory macrophages from itaconate. We find that both, non-derivatized itaconate and mesaconate dampen the glycolytic activity to a similar extent, whereas only itaconate is able to repress tricarboxylic acid cycle activity and cellular respiration. In contrast to itaconate, mesaconate does not inhibit succinate dehydrogenase. Despite their distinct impact on metabolism, both metabolites exert similar immunomodulatory effects in pro-inflammatory macrophages, specifically a reduction of interleukin (IL)-6 and IL-12 secretion and an increase of CXCL10 production in a manner that is independent of NRF2 and ATF3. We show that a treatment with neither mesaconate nor itaconate impairs IL-1β secretion and inflammasome activation. In summary, our results identify mesaconate as an immunomodulatory metabolite in macrophages, which interferes to a lesser extent with cellular metabolism than itaconate.

The Unitary Micro-Immunotherapy Medicine Interferon-γ (4 CH) Displays Similar Immunostimulatory and Immunomodulatory Effects than Those of Biologically Active Human Interferon-γ on Various Cell Types.

As a cytokine, gamma-interferon (IFN-γ) is considered a key player in the fine-tuned orchestration of immune responses. The extreme cellular sensitivity to cytokines is attested by the fact that very few of these bioactive molecules per cell are enough to trigger cellular functions. These findings can, at least partially, explain how/why homeopathically-prepared cytokines, and especially micro-immunotherapy (MI) medicines, are able to drive cellular responses. We focused our fundamental research on a unitary MI preparation of IFN-γ, specifically employed at 4 CH, manufactured and impregnated onto sucrose-lactose pillules as all other MI medicines. We assessed the IFN-γ concentration in the medium after dilution of the IFN-γ (4 CH)-bearing pillules and we evaluated in vitro drug responses in a wide range of immune cells, and in endothelial cells. Our results showed that IFN-γ (4 CH) stimulated the proliferation, the activation and the phagocytic capabilities of primary immune cells, as well as modulated their cytokine-secretion and immunity-related markers’ expression in a trend that is quite comparable with the well-recognized biological effects induced by IFN-γ. Altogether, these data provide novel and additional evidences on MI medicines, and specifically when active substances are prepared at 4 CH, thus suggesting the need for more investigations.

Altered Offspring Immunity in Maternal Parasitic Infections.

Maternal infection during pregnancy is known to alter the development and function of offspring's immune system, leading to inappropriate immune responses to common childhood infections and immunizations. Although this is an expanding field, maternal parasitic infections remain understudied. Millions of women of reproductive age are currently at risk for parasitic infection, whereas many pregnant, chronically infected women are excluded from mass drug administration due partially to a lack of resources, as well as fear of unknown adverse fetal developmental outcomes. In areas endemic for multiple parasitic infections, such as sub-Saharan Africa, there are increased rates of morbidity and mortality for various infections during early childhood in comparison with nonendemic areas. Despite evidence supporting similar immunomodulatory effects between various parasite species, there is no clear mechanistic understanding of how maternal infection reprograms offspring immunity. This brief review will compare the effects of selected maternal parasitic infections on offspring immunity.

Abstract 10791: Regeneration-Associated Cells Derived Exosomes Beneficial Than Mesenchymal Stromal Cells Derived Exosome in the Context of Myocardial Ischemia-Injury

Introduction: Vasculogenic conditioning of pro-inflammatory cells such as macrophages type 1, T cells, and primitive EPCs shifted their phenotype to pro-regenerative cells such as vasculogenic EPCs, M2 macrophages, and reg T cells, also known as regeneration-associated cells (RACs). Here, we investigated therapeutic efficacy of RAC-derived extracellular vesicles (RACev) in comparison with MSC-derived EVs (MSCev) in the context of myocardial ischemia-reperfusion injury (IRI). Methods: RACev and MSCev were isolated via ultracentrifugation. EVs quantity and quality were characterized by nanotracking analysis and CD9, CD63, Alix expression. EVs miR was sequenced and regenerative responsible miRs were validated using TargetScan and miRBase. Allogeneic immune response to the RACev and MSCev were evaluated. The function of RACev and MSCev were evaluated using HUVECs proliferation/cell-cycle assays in vitro, and repetitive (30min, d1, and d3 after IRI) systemic infusion with either RACev or MSCev in a myocardial IRI model. Results: Nanotracking analysis showed significant differences in quantity RACev vs. MSC (P= 0.03). In vitro , RACev markedly enhanced cell viability, proliferation, and migration of HUVECs in a dose-dependent manner compared to MSCev. Systemic injection of RACs (5x10 5 ) derived EVs improved cardiac functions such as ejection fraction (P= 0.005) than MSCev treated group. In histology, RACev transplanted group showed less interstitial fibrosis and enhanced capillary densities (P =0.05) compared to MSCev. These beneficial effects are coupled with significant expression of angiogenesis, anti-inflammation, cardiomyogenesis, and anti-fibrosis-related miRs in RACev but not in MSCev. In vivo bioluminescence and labeled EVs histology tracking analyses depicted preferential accumulation of RACev into the IRI myocardium (P=0.01) than MSCev. Immune phenotyping analysis confirmed similar immunomodulatory effects of MSCev and RACev. Conclusion: Repetitive systemic transplantation of RACev is superior to MSCev in terms of cardiac function enhancements via crucial angiogenesis, anti-fibrosis, anti-inflammation, cardiomyogenesis, and anti-apoptosis-related miRs delivery to the ischemic tissue.

MSCs Derived from Human Amnion Possess a Higher Proliferation Activity and Clearer Stem Cell Properties Than from Bone Marrows and They Exhibit Similar Immunosuppressive Ability In Vitro

Objectives: Human bone marrow mesenchymal stem cells (hBMSCs) have been used for the prevention and treatment of acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cells transplantation (allo-HSCT). In this study, we compared the biological characteristics and immunosuppressive activity of human amniotic mesenchymal stem cells (hAMSCs) and hBMSCs to provide experimental evidence for the potential use of hAMSCs for treating aGVHD, thus solving the problem of insufficient hBMSCs sources. Methods: hAMSCs and hBMSCs were isolated by using enzymatic digestion and Ficoll-Hypaque density gradient centrifugation, respectively, were culture-expanded in vitro to obtain the 4th-generation cells. The biological characteristics of both stem cell types were compared by morphological analysis, analysis of cell growth, cell cycle profiling, immunophenotyping, and immunofluorescence assays. An in vitro co-culture of MSCs and peripheral blood mononuclear cells (PBMCs) stimulated with phytohemagglutinin (PHA-PBMSC) was performed. The proliferation and changes of T lymphocyte subsets were measured using the Cell Counting Kit-8(CCK-8) assay and flow cytomety, respectively. The production of Interferon-γ (IFN-γ), interleukin-2 (IL-2) and IL-10 in the co-culture supernatant was determined using enzyme-linked immunosorbent assay (ELISA). Results: Both hAMSCs and hBMSCs had fibroblast-like morphology. hAMSCs could be maintained for at least 15 culture passages, but hBMSCs started to show signs of aging and remarkably reduced proliferation at 6-7 passages. There was no significant difference in the proportion of cells in G2/M phase between hAMSCs and hBMSCs (P > 0.05). Immunophenotyping revealed positive expression of CD105, CD90, and CD73 and negative expression of CD34, CD45, CD11b, CD19, and HLA-DR on the surface of both hAMSCs and hBMSCs. hAMSCs were positive for Oct-3/4, but hBMSCs were not. Both hAMSCs and hBMSCs expressed vimentin. PHA-stimulated PBMCs proliferation was inhibited by hAMSCs and hBMSCs. There were not significant differences between the inhibitory effects of the two MSCs types on PBMCs proliferation (P > 0.05). Co-culture with either hAMSCs or hBMSCs significantly increased the proportions of Treg, Th2 and Tc2 and decreased Th1 and Tc1 cell subsets in the PBMCs as compared with the PBMCs cultured alone (P < 0.05), and the changes in the PBMCs were similar between the two co-culture systems (P>0.05). In both of the two co-culture systems, IFN-γ, IL-2 production by the lymphocytes was significantly lowered (P < 0.05) and IL-10 production was significantly increased (P < 0.05) as compared with their levels in the PBMCs cultured alone; no significant difference was found in IFN-γ, IL-2 or IL-10 levels between the two co-culture systems (P > 0.05).Conclusion: The results of this study demonstrated that MSCs derived from human amnion and bone marrows have similar immunomodulatory effects on the T lymphocytes, however, hAMSCs have higher proliferation activity and clearer stem cell properties than hBMSCs, suggesting the possibility of using hAMSCs in the treatment of aGVHD after allo-HSCT. DisclosuresNo relevant conflicts of interest to declare.

The immunoregulatory functions of exosome derived from mesenchymal stem cells that are genetically modified by adeno-associated virus

目的验证天然状态间充质干细胞来源的外泌体（MSC-exosome）治疗小鼠急性移植物抗宿主病（GVHD）的效果和可能机制，探索并建立定向基因修饰MSC-exosome的方法并验证修饰后MSC-exosome的功能。方法构建小鼠急性GVHD模型，观察比较不同剂量MSC-exosome组和MSC组的生理指标评分、生存和体重减低程度；进而通过体外T细胞活化实验和体内OVA抗原特异性T细胞活化实验检测比较组间活化T细胞的增殖水平。构建重组表达载体，获得携带PD-L1和PD-L1-ITGB1的AAV2YF3突变体，进而感染人MSC，分离获得其外泌体。检测比较天然状态和修饰后MSC-exosome在体外和体内对活化T细胞的增殖水平和调节性T细胞（Treg）比例的影响。结果①MSC-exosome（300 µg×3次）和小鼠MSC（1×106×3次）均能够有效改善急性GVHD小鼠的生理指标评分、生存和体重减低程度。②相比IL-2对照组，10、25、50 µg人MSC-exosome和1×106人MSC体外共孵育处理均能够抑制活化T细胞的增殖，增殖比例分别为86.0％（IL-2）、40.0％、39.6％、42.8％和41.0％；相比PBS对照组，50、100、200 µg小鼠MSC-exosome和1×106小鼠MSC在体内均能够抑制抗原特异性活化的OT-1细胞的增殖，增殖比例分别为42.6％、33.1％、14.2％、10.6％和14.6％。③携带PD-L1和PD-L1-ITGB1的AAV2YF3突变体定向修饰人MSC-exosome的表达率分别超过40％和60％。④相比天然状态，PD-L1和PD-L1-ITGB1定向修饰后MSC-exosome在体内对抗原特异性活化的OT-1细胞具有更好的增殖抑制效果；在体外亦能明显抑制活化T细胞的增殖，并诱导提高Treg的比例。结论MSC-exosome具有与MSC相似的免疫调节作用。经过PD-L1和PD-L1-ITGB1修饰后的MSC-exosome能够有效抑制活化T细胞的增殖，并且能够诱导提高Treg的比例。

Immunomodulatory mechanism of Bacillus subtilis R0179 in RAW 264.7 cells against Candida albicans challenge

This study was aimed to explore the immunomodulatory and anti-Candida mechanisms of Bacillus subtilis (B. subtilis) R0179 in macrophages. RAW 264.7 cells were first challenged with B. subtilis R0179. B. subtilis R0179 was found to down-regulate the signals of Dectin-1, Card9, P-Iκ-Bα, Iκ-Bα, and NF-κB. Meanwhile, it reduced the levels of cytokines interleukin (IL)-1β, IL-6, IL-12, and tumor necrosis factor (TNF)-α, but increased the level of cytokine IL-10. Then RAW 264.7 cells were pretreated with B. subtilis R0179 before challenged with Candida albicans (C. albicans) or RAW 264.7 cells were co-treated with B. subtilis R0179 and C. albicans. In the presence of C. albicans, B. subtilis R0179 also showed the similar immunomodulatory effects on RAW 264.7 cells. Hence, this study provides the first insight into the immunomodulatory mechanisms of B. subtilis R0179 on the Dectin-1-related downstream signaling pathways in macrophages, which may prevent tissue damage caused by excessive pro-inflammatory response during the infection of C. albicans.

Modulatory Impact of Adipose-Derived Mesenchymal Stem Cells of Ankylosing Spondylitis Patients on T Helper Cell Differentiation.

The domination of pro-inflammatory Th subsets (Th1, Th17) is characteristic of ankylosing spondylitis (AS). Mesenchymal stem cells (MSC) were reported to normalize Th imbalance, but whether MSCs from AS adipose tissue (AS/ASCs) possess such properties is unknown. We examined AS/ASCs’ impact on Th-cell differentiation, using healthy donors ASCs (HD/ASCs) as a control. The assessment of the expression of transcription factors defining Th1 (T-bet), Th2 (GATA3), Th17 (RORc), and Treg (FoxP3) subsets by quantitative RT-PCR, the concentrations of subset-specific cytokines by ELISA, and Treg (CD4+CD25highFoxP3+) formation by flow cytometry, were performed in the co-cultures of ASCs with activated CD4+ T cells or peripheral blood mononuclear cells (PBMCs). AS/ASCs and HD/ASCs exerted similar immunomodulatory effects. Acting directly on CD4+ T cells, ASCs decreased the T-bet/GATA3 and RORc/FoxP3 ratios, diminished Treg formation, but increase IFNγ and IL-17AF production, while ASCs co-cultured with PBMCs enhanced Treg generation and reduced IFNγ release. ASCs failed to up-regulate the anti-inflammatory IL-10 and TGFβ. AS/ASCs’ impact on allogeneic and autologous PBMCs was similar. In conclusion, to shift Th differentiation to a functional anti-inflammatory direction, ASCs require accessory cell support, whereas their direct effect may be pro-inflammatory. Because ASCs neither inhibit IL-17AF nor up-regulate anti-inflammatory cytokines, their usefulness for AS patients’ treatment remains uncertain.

The novel polyclonal Ab preparation trimodulin attenuates ex vivo endotoxin-induced immune reactions in early hyperinflammation.

Sepsis is a syndrome associated with excessive inflammation. Since mortality from sepsis remains high, more laboratory research is needed to provide insight into more effective ways to use novel, potentially more beneficial agents in sepsis. We investigated the ex vivo immunomodulatory effect of a novel polyclonal Ab preparation, trimodulin, containing IgM (∼23%), IgA (∼21%) and IgG (∼56%). Using whole blood and purified PBMCs from healthy volunteers and patients with sepsis, various ex vivo investigations upon endotoxin challenge and pre- and post-trimodulin treatment were performed. Endotoxin-induced TNF-α secretion was noticeably lower with than without trimodulin, implying attenuation of the hyper-responsive state. Trimodulin also lowered TLR2, TLR4, CD11b and CD64 detection on LPS/lipoteichoic acid-stimulated monocytes. These responses were observed in cells from healthy volunteers only shortly after ex vivo endotoxin stimulation and in whole blood from patients with early-stage sepsis. Furthermore, trimodulin markedly reduced lymphocyte proliferation and release of pro- and anti-inflammatory cytokines, but did not affect phagocytosis or oxidative-burst activities of endoxin-stimulated cells. Thus, trimodulin mitigated monocyte and lymphocyte hyperinflammatory responses early after endotoxin exposure. Determining whether early in vivo administration of trimodulin will elicit similar positive immunomodulatory effects and offer a clinical benefit warrants investigation.

Oxyresveratrol ameliorates ethanol-induced gastric ulcer via downregulation of IL-6, TNF-α, NF-ĸB, and COX-2 levels, and upregulation of TFF-2 levels

Oxyresveratrol, an active ingredient of Artocarpus lakoocha, is known to possess anti-inflammatory and immunomodulatory properties. Current study investigates the immunomodulatory effect of oxyresveratrol in mouse model of ethanol-induced ulcer. Anti-ulcer effect was determined using histopathological evaluation (H&E staining) and different tests like, gastric ulcer scoring, ulcer index, total acid secretion, and gastric pH. The mRNA expression levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), nuclear factor-kappaB (NF-ĸB), cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2) and trefoil factor 2 (TFF-2) were evaluated using reverse transcription polymerase chain reaction (RT-PCR). The data showed marked percentage inhibition of erosion, hemorrhage, fibrinoid necrosis, inflammatory infiltrate, and ulcer in low (30 mg/kg b.w.) and high dose (50 mg/kg b.w.) groups of oxyresveratrol. Treatment with oxyresveratrol inhibited ulcer score and ulcer index as compared with disease control group. Oxyresveratrol significantly increased gastric pH (P < 0.001) and attenuated total acid (P < 0.001) secretion. RT-PCR analysis showed significant suppression in the mRNA expression levels of IL-6 (P < 0.001), TNF-α (P < 0.01), NF-ĸB (P < 0.001), and COX-2 (P < 0.05) in oxyresveratrol treated groups, while COX-1 expression levels were found unaltered. Treatment with oxyresveratrol significantly elevated (P < 0.01) the expression levels of cytoprotective TFF-2 levels. Similar Immunomodulatory and anti-ulcer effects were found with ranitidine treatment, which was used as a reference drug. In conclusion, oxyresveratrol possess significant anti-ulcer property which might be attributed to attenuated expression levels of IL-6, TNF-α, NF-ĸB, and COX-2 and elevated expression levels of TFF-2.

Correction: Differential regulation of innate immune cytokine production through pharmacological activation of Nuclear Factor-Erythroid-2-Related Factor 2 (NRF2) in burn patient immune cells and monocytes.

[This corrects the article DOI: 10.1371/journal.pone.0184164.].

Comparison of the Immunomodulatory Properties of Root and Leaves of Arctium Lappa (Burdock) in Vitro

Background: The roots and leaves of Arctium lappa (burdock) have been used for different therapeutic purposes, especially for diseases linked to chronic inflammation. Objectives: The present study was designed to evaluate and compare the immunomodulatory activities of root extract of burdock and leaves extract of burdock in vitro. Methods: In this experimental study, PHA- or LPS-stimulated splenocytes were treated with different concentrations of root or leaves extract of burdock and proliferation of splenocytes measured by MTT assay. The levels of IFN-γ and IL4 in the supernatants of PHA-stimulated splenocytes determined using ELISA. We also studied the effects of root and leaves extract of burdock on Nitric Oxide production by LPS-stimulated macrophages using the Griess reagent. The data were analyzed by one-way ANOVA followed by Tukey’s post-test using GraphPad Prism software, version 5.0. Results: Our findings showed that both root and leaves extract of burdock have suppressive effects on LPS-stimulated splenocytes proliferation, IL-4 secretion from PHA-stimulated splenocytes, and NO production from LPS-stimulated macrophage and stimulatory effects on PHA-stimulated splenocytes proliferation, and IFN-γ secretion from PHA-stimulated splenocytes. Although both root and leaves extract of burdock had similar immunomodulatory effects in vitro, stronger immunomodulatory effects seen in root extract of burdock. Conclusions: According to our results, we suggest that root of burdock is better option than leaves of burdock in modulation immune responses and inflammations.

Adipose-derived cellular therapies in solid organ and vascularized-composite allotransplantation.

Controlling acute allograft rejection following vascularized composite allotransplantation requires strict adherence to courses of systemic immunosuppression. Discovering new methods to modulate the alloreactive immune response is essential for widespread application of vascularized composite allotransplantation. Here, we discuss how adipose-derived cellular therapies represent novel treatment options for immune modulation and tolerance induction in vascularized composite allotransplantation. Adipose-derived mesenchymal stromal cells are cultured from autologous or allogeneic adipose tissue and possess immunomodulatory qualities capable of prolonging allograft survival in animal models of vascularized composite allotransplantation. Similar immunosuppressive and immunomodulatory effects have been observed with noncultured adipose stromal-vascular-fraction-derived therapies, albeit publication of in-vivo stromal vascular fraction cell modulation in transplantation models is lacking. However, both stromal vascular fraction and adipose derived mesenchymal stem cell therapies have the potential to effectively modulate acute allograft rejection via recruitment and induction of regulatory immune cells. To date, most reports focus on adipose derived mesenchymal stem cells for immune modulation in transplantation despite their phenotypic plasticity and reliance upon culture expansion. Along with the capacity for immune modulation, the supplemental wound healing and vasculogenic properties of stromal vascular fraction, which are not shared by adipose derived mesenchymal stem cells, hint at the profound therapeutic impact stromal vascular fraction-derived treatments could have on controlling acute allograft rejection and tolerance induction in vascularized composite allotransplantation. Ongoing projects in the next few years will help design the best applications of these well tolerated and effective treatments that should reduce the risk/benefit ratio and allow more patients access to vascularized composite allotransplantation therapy.

Differential regulation of innate immune cytokine production through pharmacological activation of Nuclear Factor-Erythroid-2-Related Factor 2 (NRF2) in burn patient immune cells and monocytes.

Burn patients suffer from immunological dysfunction for which there are currently no successful interventions. Similar to previous observations, we find that burn shock patients (≥15% Total Burn Surface Area (TBSA) injury) have elevated levels of the innate immune cytokines Interleukin-6 (IL-6) and Monocyte Chemoattractant Protein-1 (MCP-1)/CC-motif Chemokine Ligand 2(CCL2) early after hospital admission (0–48 Hours Post-hospital Admission (HPA). Functional immune assays with patient Peripheral Blood Mononuclear Cells (PBMCs) revealed that burn shock patients (≥15% TBSA) produced elevated levels of MCP-1/CCL2 after innate immune stimulation ex vivo relative to mild burn patients. Interestingly, treatment of patient PBMCs with the Nuclear Factor-Erythroid-2-Related Factor 2 (NRF2) agonist, CDDO-Me(bardoxolone methyl), reduced MCP-1 production but not IL-6 or Interleukin-10 (IL-10) secretion. In enriched monocytes from healthy donors, CDDO-Me(bardoxolone methyl) also reduced LPS-induced MCP1/CCL2 production but did not alter IL-6 or IL-10 secretion. Similar immunomodulatory effects were observed with Compound 7, which activates the NRF2 pathway through a different and non-covalent Mechanism Of Action (MOA). Hence, our findings with CDDO-Me(bardoxolone methyl) and Compound 7 are likely to reflect a generalizable aspect of NRF2 activation. These observed effects were not specific to LPS-induced immune responses, as NRF2 activation also reduced MCP-1/CCL2 production after stimulation with IL-6. Pharmacological NRF2 activation reduced Mcp-1/Ccl2 transcript accumulation without inhibiting either Il-6 or Il-10 transcript levels. Hence, we describe a novel aspect of NRF2 activation that may contribute to the beneficial effects of NRF2 agonists during disease. Our work also demonstrates that the NRF2 pathway is retained and can be modulated to regulate important immunomodulatory functions in burn patient immune cells.

A recombinant polypeptide from velvet antler of Cervus nippon Temminck exhibits similar immunomodulatory effects as its natural counterpart

Context: Velvet antler (VA) is recognized as one of the most important Chinese traditional medical herbs. To date, the immunoactivity of the single component of VA is rarely studied though its compound extracts have been well analyzed.Objective: The current study was designed to evaluate the immunomodulatory effects of a recombinant polypeptide (rVAP32) based on the VA of the sika deer by comparison with its natural counterpart (nVAP32).Materials and methods: Splenocytes proliferation and NK-cell cytotoxicity assay was evaluated by the WST-8 colorimetric method. CD4+/CD8+ cell subpopulations regulation was screened by the flowcytometry method and the Th1 or Th2-related cytokine production was measured by ELISA.Results: In vitro tests showed that both rVAP32 and nVAP32 could significantly stimulate splenocytes proliferation and enhance the NK-cell cytotoxicity and CD4+/CD8+ cell subpopulations when compared with the irrelevant peptide and blank control groups. Also, they demonstrated a significant capacity in up- and down-regulating the expression of Th1- and Th2-related cytokines, respectively. There is no statistically significant difference found between the rVAP32 tested group and nVAP32 control group.Discussion and conclusion: The results obtained herein indicate that rVAP32 has the similar immunomodulatory effects on the immune system of mice as its counterpart nVAP32 in vitro. The further test in vivo is qualified and rVAP32 is promised for developing a new biopharmaceutical product as a substitute for nVAP32.

The immunomodulatory effect of laquinimod in CNS autoimmunity is mediated by the aryl hydrocarbon receptor

Though several functional properties of laquinimod have been identified, our understanding of the underlying mechanisms is still incomplete. Since the compound elicits similar immunomodulatory effects to ligands of the aryl hydrocarbon receptor (AhR), we compared the efficacy of laquinimod in experimental autoimmune encephalomyelitis (EAE)-afflicted wild-type and AhR-deficient mice. Laquinimod failed to ameliorate clinical symptoms and leukocyte infiltration in AhR-deficient mice; however, treatment exerted neuroprotection by elevation of brain-derived neurotrophic factor (BDNF) independent of genetic profile. Thus, our data identify the AhR pathway in these mutant mice as crucial for the immunomodulatory, but not neuroprotective, efficacy of laquinimod in EAE.

Immunomodulation of Lactobacillus rhamnosus GG (LGG)-derived soluble factors on antigen-presenting cells of healthy blood donors

Lactobacillus rhamnosus GG (LGG) cells have been shown to promote type-1 immune responsiveness; however knowledge of immunomodulation of soluble factors secreted by LGG is limited. This is the first study to investigate whether LGG soluble factors promote a comparable immune responsiveness as the bacterial cells. Both treatments − LGG conditioned medium with (CM + LGG) or without (CM) LGG cells, in this study increased expression of several toll-like receptors (TLRs) in all studied cell types and antigen presentation-associated receptor HLA-DR in macrophages and “intermediate” monocytes; but decreased that of activation markers on monocytes and macrophages and production of IL-10, IL-12 and TNFα in macrophages. In co-culture with mononuclear cells, CM increased Th1-type cytokine profile but not as pronounced as CM + LGG. This study suggests that LGG soluble factors exert similar immunomodulatory effects as the intact cells, but cells may be required for optimal type-1 immune responsiveness polarizing capacity of this probiotic strain.

In Vitro Immunomodulatory Effects of Human Corneal Stromal Cells on Human Th17 Cells

Purpose: To investigate the immunomodulatory effects of human corneal stromal cells on the function of Th17 cells, in vitro.Methods: Th17 cells were co-cultured with corneal stromal cells with or without 1-methyltryptophan, 1400w dihydrochloride, interleukin (IL)-10 inhibitor, or transforming growth factor-beta (TGF-β) inhibitor. The secretion of IL-17A by Th17 cells and the upregulation of T regulatory (Treg) cell frequencies were evaluated. Transwell assays were conducted to evaluate whether the immunomodulatory effects of stromal cells were dependent on cell-to-cell contact.Results: Corneal stromal cells suppressed the secretive function of Th17 cells and upregulated Treg cell frequencies, enhancing TGF-β. Tranwell assays showed similar immunomodulatory effects of the stromal cells on the function of Th17 cells. TGF-β inhibitors attenuated the suppressive effects of the stromal cells on the function of Th17 cells, whereas other inhibitors didn't.Conclusion: It which indicates that human corneal stromal cells suppress the function of Th17 cells through a TGF-β-dependent pathway.

Colostrum and Amniotic Fluid from Different Species Exhibit Similar Immunomodulating Effects in Bacterium-Stimulated Dendritic Cells

The fetus and newborn are immunologically immature. Bioactive compounds in amniotic fluid (AF) and maternal milk therefore play a key role in the immunological development of the infant intestine. We hypothesized that colostrum and AF exert similar immunomodulatory effects on the developing immune system. Hence, bone marrow-derived murine dendritic cells (BMDCs) were co-incubated with Clostridium perfringens A or Escherichia coli Nissle 1917 and porcine, bovine, or human AF, colostrum/milk whey fractions. Interleukin (IL) 10, IL-12, IL-6, and tumor necrosis factor-? (TNF-?) production was measured. IL-12 production was reduced with all AFs and wheys, and IL-6 and TNF-? were also reduced by porcine AF. Porcine and bovine whey both reduced TNF-? production. Overall, the reductions were most pronounced with the porcine fluids. Only bovine fluids caused strong induction of IL-10. Overall, effects of AF and whey from same species were similar. Viability of stimulated BMDCs was not significantly affected by the fluids. Neutralization of IL-10, transforming growth factor ?, and epidermal growth factor (EGF) did not remove the IL-12-inhibiting effect, but EGF neutralization increased IL-10 production. Addition of EFG to DCs enhanced the bacterium-induced cytokine production contrary to the effect of AF and colostrums, ruling out EGF as the inhibitory component in the fluids.