Immunostimulatory Potential Research Articles

Immunostimulatory and anti-allergic potential of novel heterotrimeric lectin from seeds of Zizyphus mauritiana Lam

Zizyphus mauritiana Lam. seeds (ZMS) have been used medicinally as sedative or hypnotic drugs in most of Asian countries. ZMS has significant benefits to the human health. Therefore, we have evaluated immunomodulatory effect of lectin extracted from these ZMSL in both in vitro and in vivo study. Anaphylaxis is a severe life-threatening allergic reaction and Arthus reaction is deposition of immune complex and complement system activation, so we hypothesized that if ZMSL can protect these severe allergic diseases. We have studied the effect of ZMSL on macrophages and Wistar albino rats and confirmed its protective effect against anaphylaxis and Arthus reaction. Results of this study suggest ZMSL have immunostimulatory and antiallergic activity.

Water hyacinth (Eichhornia crassipes) leaves enhances disease resistance in Channa punctata from Vibrio harveyi infection

BackgroundChanna punctata, Indian spotted snakehead, has a great economic value in south and south-east Asia being an important protein source for humans. Fish cultures are affected due to various bacterial and viral infections. Vibrio harveyi is a fish pathogenic bacteria which causes several outbreaks throughout the world and leads to huge mortalities. In this study, leaves of Eichhornia crassipes (water hyacinth) were used to investigate its immunostimulatory potential in Channa punctata.ResultsThe immunostimulatory effects of water hyacinth leaves were studied in fish fed with 2.5% and 5% supplementary feed (experimental groups) in comparison to normal feed (control groups). Gas chromatography mass spectrometry (GC-MS) analysis of E. crassipes methanol extract showed presence of various components which have immunostimulatory, antioxidant, antibacterial, and anti-inflammatory activities. The antibacterial activity, antioxidant potential, and presence of phenol and flavonoids in methanol and ethanol extracts supported its use in fish feed. The healthy acclimatized fish were challenged with V. harveyi weekly. Liver function tests, alkaline phosphatase levels, and immunoglobulin content in the experimental groups were improved with respect to those in the positive control group. The spleen and head kidney were obtained at the final day of experiment, and macrophages were isolated; higher percentage of phagocytosis and phagocytic index indicated enhanced cell-mediated immune response in fish due to supplemented feed.ConclusionPlant-infused feed with leaves of E. crassipes can be recommended as a regular feed supplement to enhance fish immunity and disease resistance against the V. harveyi infection.

Immunostimulatory role of rBmHSP60 from filarial parasite Brugia malayi

Objective: To evaluate the immunostimulatory potential of cross-reactive molecule heat shock protein 60 (HSP60) of filarial parasite Brugia malayi and Leishmania donovani. Methods: HSP60 of Brugia malayi (BmHSP60) was amplified using gene-specific primer, cloned in pTriEx4 vector, expressed in BL21-DE3 cells, and recombinant HSP60 (rHSP60) of ~65 kDa was purified by affinity chromatography using Ni-NTA column. The recombinant protein was desalted by the dialysis membrane, and the presence of endotoxin level was determined by Limulus amebocyte lysate assay. The recombinant protein was tested for cell proliferation, nitric oxide release, expression of Th1 and Th2 cytokines, and transcription factors (STATs) in vitro using murine macrophage cell line (J774A.1). Results: Higher cell proliferation indicated that BmHSP60 had immunostimulatory potential. rBmHSP60 exposure upregulated the expression of iNOS, STAT1, STAT4, Th1 cytokines (IFN–γ, TNF–α, IL-12), and nitric oxide release. In addition, no remarkable change was observed in the expression of IL-6, IL-10, and STAT3 in macrophage cell line J774A.1. The ELISA analysis showed the levels of IFN-γ, TNF-α, and IL-12 were upregulated while IL-10 level was downregulated, revealing that BmHSP60 triggered a Th1 immune response. Conclusions: Our study demonstrates that rBmHSP60 has immunogenic properties which effectively enhances the Th1 type immune responses, and can be used as an immunoprophylactic agent against leishmaniasis. Furthermore, in vivo studies are in progress to determine the protective role of rBmHSP60 against Leishmania donovani infection in a mouse model.

Oncolytic virotherapy – Forging its place in the immunomodulatory paradigm for Multiple Myeloma

The treatment focus for multiple myeloma (MM) has recently pivoted towards immune modulating strategies, with T-cell redirection therapies currently at the forefront of drug development. Yet, despite this revolution in treatment, MM remains without a sustainable cure. At the same time, tremendous advancement has been made in recombinant and gene editing techniques for oncolytic viruses (OV), which have increased their tumor specificity, improved safety, and enhanced the oncolytic and immunostimulatory potential. These breakthrough developments in oncolytic virotherapy have opened new avenues for OVs to be used in combination with other immune-based therapies such as checkpoint inhibitors, chimeric antigen receptor T-cells (CAR-T) and bispecific T-cell engagers. In this review, the authors place the spotlight on systemic oncolytic virotherapy as an adaptable immunotherapeutic for MM, highlight the unique mechanism of OVs in activating the immune-suppressive marrow microenvironment, and lastly showcase the OV platforms and the promising combination strategies in the pipeline for MM.

A review article on immunostimulatory potential in Moringa oleifera

The present review study was conducted during the year 2020 at Chandigarh Group of Colleges, Jhanjeri, Mohali. The study results indicated that Moriga is one of the nutritious source among all the food sources. The different phyto-chemicals play a vital role to improve the human immune system. The different chemical constituents present in different parts of the plant having nutraceutical property and can be utilized in different ways. Moringa may be a great alternate source of Vitamins, Protein, Fiber, Carbohydrates and different Minerals. Some of the severe diseases like Cancer, Diabetics, Cerebral ischemia etc. can be completely avoid by consuming Moringa oleifera.

Administration of Different Derivatives of Oliveria decumbens Improves Innate Immunity of Nile Tilapia (Oreochromis niloticus) Without Affecting Fish Growth and Blood Biochemical Parameters

The objectives of this study were to investigate the effects of Oliveria decumbens as a medicinal herb with reported immunostimulatory potential on the growth, immunity status and health of Nile tilapia (Oreochromis niloticus). In an eight- week trial, fish (45 ± 5 g) were randomly divided into 13 treatments as follows: in 10 treatments fish were fed on diets containing 0 (control), 0.01, 0.1 and 1% of O. decumbens extract and essential oil and their 1:1 combinations. Also, in three treatments fish received plant hydrolate at doses of 312.5, 625 and 1250 ppm as bath treatment. At the end of experiment, blood samples were taken for immunological and biochemical measurements. All treated fish showed comparable growth performance to those received control diet. The highest levels of respiratory burst activity were observed in extract 1% group. Similarly the serum lysozyme levels were significantly affected by essential oil and extract supplemented diets. Fish received plant hydrolate at dose 312.5 ppm showed the highest significant protein level. Also, the globulin levels were increased in group fed on diet containing extract 1%. Plant supplementation had no negative effects on the fish health biochemical indices including cholesterol, triglyceride, alkaline phosphatase and aminotransferase enzymes. In conclusion, the results of this study showed that application of O. decumbens derivatives improved the immunity of Nile tilapia without adverse effects on fish growth and health.

Crude polysaccharide from the milky mushroom, Calocybe indica, modulates innate immunity of macrophage cells by triggering MyD88-dependent TLR4/NF-κB pathway.

Calocybe indica is a famous nutritious food in Asian countries and one of the most widely cultivated mushrooms in the world. Here, we have isolated crude polysaccharides from this mushroom, characterized it and investigated its antioxidant and immunostimulatory potential. The polysaccharide was chemically characterized by spectrophotometry, FTIR and high-performance thin layer chromatography and tested its antioxidant potential by in vitro assays. Immunomodulatory activity and its underlying signalling process were ascertained in RAW 264.7 cells. The polysaccharide consisted of D-glucose (β-linked sugars), D-mannose and D-galactose, where backbone was organized in random coil structure. Preliminary investigation of the bioactivity of the polysaccharide revealed its antioxidant potential. The polysaccharide could noticeably induce phagocytic activity and production of immune mediators in macrophage cells. The polysaccharide was found to enhance the expression of pro-inflammatory cytokines and activate NF-κB signalling pathway by overexpressing MyD88, Iκ-Bα and NF-κB. Further studies indicated the polysaccharide binds to the toll-like receptor 4 to manifest its immunostimulatory activity in macrophage cells. Our findings indicate potential therapeutic properties of the crude polysaccharide of C. indica which might provide the means to treat various radical induced and immunodeficiency disorders in the days to come.

Evaluation of the Antibacterial Activity, Acute Toxicity and Immuno-stimulatory Potential of Adenodolichos paniculatus root Extract

Medicinal plants have been used in the treatment of illnesses from time immemorial. In Nigeria today a large population of people still depend on herbal medicines for their health needs. The medicinal plant, Adenodolichos paniculatus has been used traditionally as a remedy for mouth and throat infections in Pushit community of Plateau state, Nigeria. Five solvents (hexane, chloroform, ethyl acetate, methanol and water) were used sequentially for the isolation of bioactive compounds from the root of the plant Adenodolichos paniculatus and concentrated on rotary vacuum evaporator. Concentrated extracts were tested for their antibacterial activities against Streptococcus pyogenes, Staphylococcus aureus, Pseudomonas aeruginosa and E. coli by agar-well diffusion method. Ofloxacin was the referenced drug. Chloroform root extract was more potent than the remaining four extracts. The extract at 100 mg/ml was active against only three (3) of the four (4) pathogens tested namely Streptococcus pyogenes, Staphylococcus aureus and Pseudomonas aeruginosa with zones of inhibition measuring 25.00, 21.00 and 11.75 mm respectively. The minimum inhibitory concentration of the extract against the test organisms were 1,560, 6,250 and 25,000 µg/ml respectively. The minimum bactericidal concentration was 3,120, 12,500 and 50,000 µg/ml against the three organisms. The extract was found to be nontoxic with LD50 of above 2000 (mg//kg body weight) when administered orally in mice. The plant extract has immunostimulatory activity with a significant increase (p<0.05) in white blood cell proliferation when administered into mice at doses of 50, 25 and 100 mg/kg. Conclusion: The result of this investigation supports the use of this plant for the traditional remedy of mouth and throat infections possibly caused by these test organisms. The extract can serve as lead in phytomedicine development for the treatment of throat related infections.

O-Polysaccharide Plays a Major Role on the Virulence and Immunostimulatory Potential of Aggregatibacter actinomycetemcomitans During Periodontal Infection.

Aggregatibacter actinomycetemcomitans is a Gram-negative oral bacterium with high immunostimulatory and pathogenic potential involved in the onset and progression of periodontitis, a chronic disease characterized by aberrant immune responses followed by tooth-supporting bone resorption, which eventually leads to tooth loss. While several studies have provided evidence related to the virulence factors of A. actinomycetemcomitans involved in the host cell death and immune evasion, such as its most studied primate-specific virulence factor, leukotoxin, the role of specific lipopolysaccharide (LPS) domains remain poorly understood. Here, we analyzed the role of the immunodominant domain of the LPS of A. actinomycetemcomitans termed O-polysaccharide (O-PS), which differentiates the distinct bacterial serotypes based on its antigenicity. To determine the role of the O-PS in the immunogenicity and virulence of A. actinomycetemcomitans during periodontitis, we analyzed the in vivo and in vitro effect of an O-PS-defective transposon mutant serotype b strain, characterized by the deletion of the rmlC gene encoding the α-L-rhamnose sugar biosynthetic enzyme. Induction of experimental periodontitis using the O-PS-defective rmlC mutant strain resulted in lower tooth-supporting bone resorption, infiltration of Th1, Th17, and Th22 lymphocytes, and expression of Ahr, Il1b, Il17, Il23, Tlr4, and RANKL (Tnfsf11) in the periodontal lesions as compared with the wild-type A. actinomycetemcomitans strain. In addition, the O-PS-defective rmlC mutant strain led to impaired activation of antigen-presenting cells, with less expression of the co-stimulatory molecules CD40 and CD80 in B lymphocytes and dendritic cells, and downregulated expression of Tnfa and Il1b in splenocytes. In conclusion, these data demonstrate that the O-PS from the serotype b of A. actinomycetemcomitans plays a key role in the capacity of the bacterium to prime oral innate and adaptive immune responses, by triggering the Th1 and Th17-driven tooth-supporting bone resorption during periodontitis.

Use of human peripheral blood mononuclear cells to define immunological properties of nucleic acid nanoparticles.

This protocol assesses proinflammatory properties of nucleic acid nanoparticles (NANPs) using a validated preclinical model, peripheral blood mononuclear cells (PBMCs), that is highly predictive of cytokine responses. The experimental procedure details the preparation of pyrogen-free NANPs, isolation of PBMCs from freshly collected human blood, and analysis of characteristic biomarkers (type I and III interferons) produced by PBMCs transfected with NANPs. Although representative NANPs with high and low immunostimulatory potential are used as standards throughout the procedure, this protocol can be adapted to any NANPs or therapeutic nucleic acids, irrespective of whether they are carrier based or carrier free; additional cytokine biomarkers can also be included. We test several commercial platforms and controls broadly accessible to the research community to quantify all biomarkers in either single- or multiplex format. The continuous execution of this protocol takes <48 h; when immediate analysis is not feasible, single-use aliquots of the supernatants can be frozen and stored (-20 °C; 12 months).

The immunorecognition, subcellular compartmentalization, and physicochemical properties of nucleic acid nanoparticles can be controlled by composition modification.

Nucleic acid nanoparticles (NANPs) have become powerful new platforms as therapeutic and diagnostic tools due to the innate biological ability of nucleic acids to identify target molecules or silence genes involved in disease pathways. However, the clinical application of NANPs has been limited by factors such as chemical instability, inefficient intracellular delivery, and the triggering of detrimental inflammatory responses following innate immune recognition of nucleic acids. Here, we have studied the effects of altering the chemical composition of a circumscribed panel of NANPs that share the same connectivity, shape, size, charge and sequences. We show that replacing RNA strands with either DNA or chemical analogs increases the enzymatic and thermodynamic stability of NANPs. Furthermore, we have found that such composition changes affect delivery efficiency and determine subcellular localization, effects that could permit the targeted delivery of NANP-based therapeutics and diagnostics. Importantly, we have determined that altering NANP composition can dictate the degree and mechanisms by which cell immune responses are initiated. While RNA NANPs trigger both TLR7 and RIG-I mediated cytokine and interferon production, DNA NANPs stimulate minimal immune activation. Importantly, incorporation of 2′F modifications abrogates RNA NANP activation of TLR7 but permits RIG-I dependent immune responses. Furthermore, 2′F modifications of DNA NANPs significantly enhances RIG-I mediated production of both proinflammatory cytokines and interferons. Collectively this indicates that off-target effects may be reduced and/or desirable immune responses evoked based upon NANPs modifications. Together, our studies show that NANP composition provides a simple way of controlling the immunostimulatory potential, and physicochemical and delivery characteristics, of such platforms.

Administration of Extensive Hydrolysates From Caseins and Lactobacillus rhamnosus GG Probiotic Does Not Prevent Cow's Milk Proteins Allergy in a Mouse Model.

BackgroundEarly nutrition may influence the development of food allergies later in life. In the absence of breastfeeding, hydrolysates from cow’s milk proteins (CMP) were indicated as a prevention strategy in at risk infants, but their proof of effectiveness in clinical and pre-clinical studies is still insufficient. Thanks to a validated mouse model, we then assessed specific and nonspecific preventive effects of administration of extensive hydrolysates from caseins (eHC) on the development of food allergy to CMP. The additional nonspecific effect of the probiotic Lactobacillus GG (LGG), commonly used in infant formula, was also assessed.MethodsGroups of young BALB/cByJ female mice were pretreated by repeated gavage either with PBS (control mice), or with PBS solution containing non-hydrolyzed milk protein isolate (MPI), eHC or eHC+LGG (eq. of 10 mg of protein/gavage). All mice were then experimentally sensitized to CMP by gavage with whole CM mixed with the Th2 mucosal adjuvant Cholera toxin. All mice were further chronically exposed to cow’s milk. A group of mice was kept naïve. Sensitization to both caseins and to the non-related whey protein β-lactoglobulin (BLG) was evaluated by measuring specific antibodies in plasma and specific ex vivo Th2/Th1/Th17 cytokine secretion. Elicitation of the allergic reaction was assessed by measuring mMCP1 in plasma obtained after oral food challenge (OFC) with CMP. Th/Treg cell frequencies in gut-associated lymphoid tissue and spleen were analyzed by flow cytometry at the end of the protocol. Robust statistical procedure combining non-supervised and supervised multivariate analyses and univariate analyses, was conducted to reveal any effect of the pretreatments.ResultsPBS pretreated mice were efficiently sensitized and demonstrated elicitation of allergic reaction after OFC, whereas mice pretreated with MPI were durably protected from allergy to CMP. eHC+/-LGG pretreatments had no protective effect on sensitization to casein (specific) or BLG (non-specific), nor on CMP-induced allergic reactions. Surprisingly, eHC+LGG mice demonstrated significantly enhanced humoral and cellular immune responses after sensitization with CMP. Only some subtle changes were evidenced by flow cytometry.ConclusionNeither specific nor nonspecific preventive effects of administration of casein-derived peptides on the development of CMP food allergy were evidenced in our experimental setup. Further studies should be conducted to delineate the mechanisms involved in the immunostimulatory potential of LGG and to clarify its significance in clinical use.

In vitro cytomodulatory and immunomodulatory effects of bovine colostrum whey protein hydrolysates

SummaryBovine colostrum possesses biological compounds involved in the development of the newborn. Among them, the proteins have drawn attention as a source of bioactive peptides. This study shows the in vitro cytotoxic and immunostimulatory potential of two colostrum whey protein (CWP) hydrolysates obtained by in vitro digestion with pepsin and pancreatin. MTT cell viability, apoptosis induction, polymorphonuclear proliferation and phagocytic activity assays were performed. Treatment with the hydrolysates induced a significant decrease in the viability of MDA‐MB‐231 cell lines due to apoptosis and also a significant increase in the proliferation of blood mononuclear cells. It could also be observed that for the RM‐1 and PC‐3 prostate cancer cell lines and for the two times of exposure (24 and 48 h), the hydrolysate H1 is significantly more cytotoxic than CWP. These results showed the potential of bovine CWP and its hydrolysates for the treatment of chronic diseases such as cancer.

Targeted Theranostic Nano Vehicle Endorsed with Self‐Destruction and Immunostimulatory Features to Circumvent Drug Resistance and Wipe‐Out Tumor Reinitiating Cancer Stem Cells

The downsides of conventional cancer monotherapies are profound and enormously consequential, as drug-resistant cancer cells and cancer stem cells (CSC) are typically not eliminated. Here, a targeted theranostic nano vehicle (TTNV) is designed using manganese-doped mesoporous silica nanoparticle with an ideal surface area and pore volume for co-loading an optimized ratio of antineoplastic doxorubicin and a drug efflux inhibitor tariquidar. This strategically framed TTNV is chemically conjugated with folic acid and hyaluronic acid as a dual-targeting entity to promote folate receptor (FR) mediated cancer cells and CD44 mediated CSC uptake, respectively. Interestingly, surface-enhanced Raman spectroscopy is exploited to evaluate the molecular changes associated with therapeutic progression. Tumor microenvironment selective biodegradation and immunostimulatory potential of the MSN-Mn core are safeguarded with a chitosan coating which modulates the premature cargo release and accords biocompatibility. The superior antitumor response in FR-positive syngeneic and CSC-rich human xenograft murine models is associated with a tumor-targeted biodistribution, favorable pharmacokinetics, and an appealing bioelimination pattern of the TTNV with no palpable signs of toxicity. This dual drug-loaded nano vehicle offers a feasible approach for efficient cancer therapy by on demand cargo release in order to execute complete wipe-out of tumor reinitiating cancer stem cells.

Abstract A12: Bladder cancer-derived extracellular vesicles and immune activation

Abstract Background: Bladder tumors occur on the luminal surface of the bladder, and cancer cells and cellular components are shed in urine, which permits the isolation of an enriched population of bladder cancer-specific extracellular vesicles (EVs). EVs contain proteins, miRNA, mRNA, and cellular metabolites, which may serve as stimulators of immune cells. The objective of this study was to evaluate the immune potential of tumor-derived EVs (TEVs) produced on exposure to BCG using a murine bladder cancer cell line and immune cells. Methods: TEVs were isolated by differential high-speed centrifugation of cell culture supernatants. They were confirmed to be extracellular vesicles by electron microscopy and size measurement using nanoparticle tracking analysis. Unstimulated murine bladder cancer cells (MB49) were used to prepare TEV1. The cells were then exposed to BCG Tokyo (MOI 4) and TEVs were isolated after 2 hours (TEV2). BCG was then washed off and the cells were allowed to grow in complete media for 2 days before isolation of the third sample, TEV3. EVs from BCG (BEV) were collected to serve as a control. Bone marrow-derived cells were obtained from C57BL/6J mice to generate dendritic cells (DC) and neutrophils. These cells were untreated or treated with 1ug of TEVs or BEV or BCG (MOI 5) for 24 hours. The supernatants were analyzed for cytokine production and the DCs and neutrophils were analyzed for activation markers by flow cytometry. The institutional animal care and use committee approved the use of mice. Results: DCs treated with TEV2 had significantly higher expression of CD86 (at similar levels when DCs were treated with BCG) and MHC2, both of which enhanced the DC's capacity to present antigens to T cells, when compared to TEV1 or TEV3. While neutrophils induced by BCG expressed significantly higher levels of costimulatory molecules CD80, CD83, CD86, and MHC2, TEV2 induced significant expression of CD80 and CD86. BCG caused a massive release of proinflammatory cytokines (IL6, TNFa, and IL12p70) and the anti-inflammatory cytokine IL10. Cytokine production induced by TEV2 was not as high as that induced by direct BCG stimulation, but it was significantly higher than that produced by untreated cells or TEV1 and TEV3. Unlike BCG, TEV2 did not significantly stimulate IL10 production. Conclusion: Our results demonstrate that tumor cells treated with BCG for 2 hours produce EVs that are able to cause significant modulation of DCs and neutrophils. TEVs generated during BCG immunotherapy have immunostimulatory potential. Therefore, TEVs represent a promising source of material to increase immune activation and efficacy of BCG therapy. This could reduce the number of instillations and adverse effects associated with BCG therapy. Citation Format: Sin Mun Tham, Kesavan Esuvaranathan, Ratha Mahendran. Bladder cancer-derived extracellular vesicles and immune activation [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2019 May 18-21; Denver, CO. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(15_Suppl):Abstract nr A12.

Dietary supplementation of marine yeast Yarrowia lipolytica modulates immune response in Litopenaeus vannamei

The immunostimulatory potential of the marine yeast Yarrowia lipolytica (D1 and N6 strains) administered orally was evaluated in the white shrimp Litopenaeus vannamei. Yeasts and commercial glucans were mixed with a commercial feed to formulate diets with a 1.1% concentration of immunostimulants. The shrimp were fed daily for a period of 21 days. Weekly determinations were performed for immunological parameters in hemolymph, such as total hemocyte count (THC), lysozyme activity (LYZ), prophenoloxidase activity, antioxidant enzymatic activities (superoxide dismutase [SOD], catalase [CAT], and peroxidases), and bactericidal activity against Vibrio parahaemolyticus. Expression profiles of penaeidin (PEN), lysozyme (LYZ), and prophenoloxidase (proPO) immune genes were evaluated in hemocytes. In general, an increase in the immune parameters was observed in shrimp fed yeast diet compared to glucan and the control diets. Yarrowia lipolytica, especially strain N6, provided maximum immunostimulatory effects evidenced by the increase of immune parameters (THC, LYZ, SOD, CAT) and gene expression profile. In conclusion, this study demonstrated that Y. lipolytica had immunostimulatory effects and increased bactericidal activity in L. vannamei hemocytes against V. parahaemolyticus. These findings open the path for the potential application of Y. lipolytica-based immunostimulant for shrimp aquaculture.

Immunostimulatory Potential of Natural Compounds and Extracts: A Review

The proper functioning of human immune system is essential for organism survival against infectious, toxic and oncogenic agents. The present study aimed to describe the scientific evidence regarding the immunomodulatory properties of the main micronutrients and specific phytochemicals. Plants of food interest have the ability to dynamically affect the immune system through particular molecules. Plant species, type of compounds and biological effects were herein reviewed mainly focusing on plants which are not commonly used in food supplements. Several efficient phytoproducts showed significant advantages compared to synthetic immunomodulators, being good candidates for the development of immunotherapeutic drugs.

Wheat (Triticum aestivum L.) Breeding from 1891 to 2010 Contributed to Increasing Yield and Glutenin Contents but Decreasing Protein and Gliadin Contents.

Epidemiologic studies suggest an increasing prevalence of celiac disease and non-celiac gluten/wheat sensitivity. With wheat proteins being the main triggers, changes in wheat protein composition are discussed as a potential cause. The goals of breeding toward increased yield and resistance might have inadvertently contributed to a higher immunostimulatory potential of modern wheat cultivars compared to old wheat cultivars. Therefore, agronomic characteristics, protein content, and gluten composition of 60 German winter wheat cultivars first registered between 1891 and 2010 grown in 3 years were analyzed. While plant height and spike density decreased over time, yield and harvest index increased. The protein and gliadin contents showed a decreasing trend, whereas glutenin contents increased, but there were no changes in albumin/globulin and gluten contents. Overall, the harvest year had a more significant effect on protein composition than the cultivar. At the protein level, we found no evidence to support an increased immunostimulatory potential of modern winter wheat.

RNA Origami Nanostructures for Potent and Safe Anticancer Immunotherapy.

Rapid developments in nucleic acid nanotechnology have enabled the rational design and construction of self-assembling DNA and RNA nanostructures that are highly programmable. We recently developed a replicable single-stranded RNA origami (RNA-OG) technology that allows a long RNA molecule to be programmed to self-assemble into nanostructures of various shapes. Here, we show that such RNA-OG is highly stable in serum/plasma, and we thus exploited its immunostimulatory potential. We demonstrated that the RNA-OG stimulates a potent innate response primarily through a Toll-like receptor 3 (TLR3) pathway. In a murine peritoneal metastatic colon cancer model, intraperitoneally injected RNA-OG induced significant tumor retardation or regression by activating NK- and CD8-dependent antitumor immunity and antagonizing the peritoneal immunosuppressive environment. Unlike polyinosinic/polycytidylic acid (PolyIC), a well-known double-stranded RNA analogue, the RNA-OG treatment did not cause a high level of type-I interferons in the blood nor apparent toxicity upon its systemic administration in the animals. This work establishes the function of RNA-OG as a potent line of TLR3 agonists that are safe and effective for cancer immunotherapy.

Immunostimulatory Potential of MoS2 Nanosheets: Enhancing Dendritic Cell Maturation, Migration and T Cell Elicitation.

BackgroundDue to their extraordinary physical and chemical properties, MoS2 nanosheets (MSNs) are becoming more widely used in nanomedicine. However, their influence on immune systems remains unclear.Materials and MethodsTwo few-layered MSNs at sizes of 100–250 nm (S-MSNs) and 400–500 nm (L-MSNs) were used in this study. Bone marrow-derived dendritic cells (DCs) were exposed to both MSNs at different doses (0, 8, 16, 32, 64, 128 µg/mL) for 48 h and subjected to analyses of surface marker expression, cytokine secretion, lymphoid homing and in vivo T cell priming.ResultsDifferent-sized MSNs of all doses did not affect the viability of DCs. The expression of CD40, CD80, CD86 and CCR7 was significantly higher on both S-MSN- and L-MSN-treated DCs at a dose of 128 μg/mL. As the dose of MSN increased, the secretion of IL-12p70 remained unchanged, the secretion of IL-1β decreased, and the production of TNF-α increased. A significant increase in IL-6 was observed in the 128 µg/mL L-MSN-treated DCs. In particular, MSN treatment dramatically improved the ex vivo movement and in vivo homing ability of both the local resident and blood circulating DCs. Furthermore, the cytoskeleton rearrangement regulated by ROS elevation was responsible for the enhanced homing ability of the MSNs. More robust CD4+ and CD8+ T cell proliferation and activation (characterized by high expression of CD107a, CD69 and ICOS) was observed in mice vaccinated with MSN-treated DCs. Importantly, exposure to MSNs did not interrupt LPS-induced DC activation, homing and T cell priming.ConclusionFew-layered MSNs ranging from 100 to 500 nm in size could play an immunostimulatory role in enhancing DC maturation, migration and T cell elicitation, making them a good candidate for vaccine adjuvants. Investigation of this study will not only expand the applications of MSNs and other new transition metal dichalcogenides (TMDCs) but also shed light on the in vivo immune-risk evaluation of MSN-based nanomaterials.