Immune Stimulatory Properties Research Articles

Utilisation of dates for the formulation of functional food product

The date palm (Phoenix dactylifera L.) is one of the popular perennial fruit tree grown around the world, particularly in West Asia and North Africa. Its fruit (date) is popular all throughout the world. The tree is well-known for its beneficial health benefits and is regarded as a high-value commercial fruit crop in the worldwide market. Dates are a high-quality, low-cost source of macro- and micronutrients. Date fruits are mostly composed of carbohydrates, including soluble sugars and dietary fibre, with modest quantities of fats and proteins. Dates also poses antioxidant, anti-mutagenic, anti-microbial, hepato-protective, gastro-protective, anti-inflammatory, immune-stimulatory and anti-cancererous properties. Despite the lack of commercialization of many varieties, date palm fruits are important marketable crops. An explosion of interest in the numerous health benefits of dates in recent years has resulted in numerous in vitro and animal studies, as well as the identification and quantification of various classes of phytochemicals. Food scientists and the food industry have created a vast variety of food products made from dates, including paste, syrup, juice, sugar, jam, jelly, butter, chocolates, condiments, pickled dates, oils, coffee etc. This review includes updated information concerning the composition of several varieties of dates, as well as the possible health advantages of Phoenix dactylifera and processed items made from date fruit.

Immune-Boosting Plants Used in Turkish Folk Medicine and their Potential against COVID-19

Having suffered from a global epidemic of Covid19 mutants and variants, the entire world has turned its attention and hope to natural treatments that can keep the immune system active and strong. Recent research on some herbal preparations promises high hopes that they may be potential candidates for developing effective and safe immune-boosting drugs. Considering that Turkey has the richest flora of Europe and the Middle East and has an invaluable ethnomedical background, it offers a treasure-worthy potential for serious researchers. 128 plant taxa belonging to 41 families were determined from 99 works carried out in different regions of Turkey. Of these, only 60 (46.9%) were found to be responded to in experimental studies worldwide on immune system enhancement and other similar activities. Quercetin (10.3%), β-glucan, and catechin (5.1%) emerged as the most common active substances among 39 identified active substances. Urtica dioica (39.4%), Rosa canina (34.3%), and Punica granatum (17.2%) were recorded as the most used plants in Turkey. In addition, Phoenix dactylifera has appeared as the taxon with having more active compounds in the literature. This study presents the first national inventory of immune-boosting plants that are regularly used in traditional Turkish medicine, not only against Covid-19 but for the prevention and treatment of epidemic diseases in general. We present 68 (53.1%) plants whose effective immune-stimulating and strengthening properties have been confirmed in experimental studies in the world literature as ready-made material to the relevant pharmacological sectors.

STING activation promotes robust immune response and NK cell–mediated tumor regression in glioblastoma models

Immunotherapy has had a tremendous impact on cancer treatment in the past decade, with hitherto unseen responses at advanced and metastatic stages of the disease. However, the aggressive brain tumor glioblastoma (GBM) is highly immunosuppressive and remains largely refractory to current immunotherapeutic approaches. The stimulator of interferon genes (STING) DNA sensing pathway has emerged as a next-generation immunotherapy target with potent local immune stimulatory properties. Here, we investigated the status of the STING pathway in GBM and the modulation of the brain tumor microenvironment (TME) with the STING agonist ADU-S100. Our data reveal the presence of STING in human GBM specimens, where it stains strongly in the tumor vasculature. We show that human GBM explants can respond to STING agonist treatment by secretion of inflammatory cytokines. In murine GBM models, we show a profound shift in the tumor immune landscape after STING agonist treatment, with massive infiltration of the tumor-bearing hemisphere with innate immune cells including inflammatory macrophages, neutrophils, and natural killer (NK) populations. Treatment of established murine intracranial GL261 and CT-2A tumors by biodegradable ADU-S100-loaded intracranial implants demonstrated a significant increase in survival in both models and long-term survival with immune memory in GL261. Responses to treatment were abolished by NK cell depletion. This study reveals therapeutic potential and deep remodeling of the TME by STING activation in GBM and warrants further examination of STING agonists alone or in combination with other immunotherapies such as cancer vaccines, chimeric antigen receptor T cells, NK therapies, and immune checkpoint blockade.

Abstract 5205: VG2025: Transcriptional and translational dually regulated (TTDR) oncolytic virus for cancer immunotherapy

Abstract VG2025 is a genetically modified HSV-1 oncolytic virus (OV) wherein the transcription and translation of critical viral genes are regulated to enhance virus safety and promote tumor specific virus replication without attenuating the virus. The TTDR design is based on transcriptional control of the essential HSV-1 immediate-early protein ICP27 using a tumor specific CEA promoter, coupled with translational control of the key neurovirulence factor ICP34.5 accomplished by introducing multiple tandem copies of miRNA binding sites for miR-124 and miR-143 which are downregulated in tumors but abundantly expressed in normal brain tissues. The anti-tumor and immune stimulatory properties of VG2025 are also enhanced by expressing a pair of cytokines (IL-12 and IL-15/IL-15Rα complex) that are linked by 2A cleavage peptides and controlled by the tumor specific promoter CXCR4. The dual regulation strategy was verified in vitro and shown to be highly selective. Growth of VG2025 virus correlated with CEA expression in a panel of cell lines including normal cells and a selection of tumor cells with varying CEA levels. miRNA-mediated regulation of ICP34.5 expression was validated by transfecting HEK293t cells with miR-124 and miR-143 precursors, followed by infection with VG2025. RT-qPCR testing revealed a significant decrease in the levels of viral ICP34.5 mRNA when the miRNA was present. VG2025 virus replication was evaluated in a variety of human tumor cell lines, and two cell lines representing NSCLC and pancreatic cancer (A549 and BxPC3, respectively) were selected for testing in a humanized mouse model, where VG2025 displayed strong in vivo anti-tumor efficacy. The immune stimulatory characteristics of VG2025, and the bioactivity of expressed cytokines, were demonstrated in a syngeneic mouse colorectal tumor model (CT26) and lymphoma model (A20) using VG2026, which is an identical mouse specific variant of VG2025 that expresses murine IL-12 in place of human IL-12. In addition to a significant reduction in the growth of injected tumors, we observed a clear abscopal effect on contralateral tumors, indicating a robust anti-tumor immune response with an immune memory effect. Due to the tendency of HSV-1 to establish latency which may lead to neurotoxicity and potential safety concerns, we performed a safety pharmacology study by subcutaneously inoculating immunocompetent DBA mice with VG2025. Absence of neurotoxicity was further verified by more aggressive routes of administration, including via the nose and by corneal scarification. The risk of VG2025 reactivation from a latent state was evaluated in a two-step study: latent infection in the CNS was first established and confirmed, then the trigeminal ganglia were removed and co-cultured with Vero cells to detect reactivation-related cytopathic effects. All studies showed that VG2025 was safe with no reactivation events detected. Citation Format: Yanal Murad, Guoyu Liu, I-Fang Lee, Zahid Delwar, Dmitry Chouljenko, Will Liu, Jun Ding, Luke Bu, Ronghua Zhao, William Jia. VG2025: Transcriptional and translational dually regulated (TTDR) oncolytic virus for cancer immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5205.

Extracellular vimentin mimics VEGF and is a target for anti-angiogenic immunotherapy

Anti-angiogenic cancer therapies possess immune-stimulatory properties by counteracting pro-angiogenic molecular mechanisms. We report that tumor endothelial cells ubiquitously overexpress and secrete the intermediate filament protein vimentin through type III unconventional secretion mechanisms. Extracellular vimentin is pro-angiogenic and functionally mimics VEGF action, while concomitantly acting as inhibitor of leukocyte-endothelial interactions. Antibody targeting of extracellular vimentin shows inhibition of angiogenesis in vitro and in vivo. Effective and safe inhibition of angiogenesis and tumor growth in several preclinical and clinical studies is demonstrated using a vaccination strategy against extracellular vimentin. Targeting vimentin induces a pro-inflammatory condition in the tumor, exemplified by induction of the endothelial adhesion molecule ICAM1, suppression of PD-L1, and altered immune cell profiles. Our findings show that extracellular vimentin contributes to immune suppression and functions as a vascular immune checkpoint molecule. Targeting of extracellular vimentin presents therefore an anti-angiogenic immunotherapy strategy against cancer.

The HLA-G immune checkpoint: a new immuno-stimulatory role for the α1-domain-deleted isoform.

The heterogeneity of cancer cells, in part maintained via the expression of multiple isoforms, introduces significant challenges in designing effective therapeutic approaches. In this regard, isoforms of the immune checkpoint HLA-G have been found in most of the tumors analyzed, such as ccRCC, the most common human renal malignancy. In particular, HLA-G∆α1, which is the only HLA-G isoform described that lacks the α1 extracellular domain, has been newly identified in ccRCC and now here in trophoblasts. Using a cellular model expressing HLA-G∆α1, we have uncovered its specific and overlapping functional roles, relative to the main HLA-G isoform, i.e., the full-length HLA-G1. We found that HLA-G∆α1 has several particular features: (i) although possessing the α3 domain, it does not associate with β2-microglobulin; (ii) it may not present peptides to T cells due to absence of the peptide-binding groove; and (iii) it exerts immune-stimulatory activity towards peripheral blood NK and T cells, while all known isoforms of HLA-G are immune-inhibitory checkpoint molecules. Such immune-stimulatory properties of HLA-G∆α1 on the cytotoxic function of peripheral blood NK cells are individual dependent and are not exerted through the interaction with the known HLA-G receptor, ILT2. Importantly, we are faced here with a potential antitumor effect of an HLA-G isoform, opposed to the pro-tumor properties described for all other HLA-G isoforms, which should be taken into account in future therapeutic designs aimed at blocking this immune checkpoint.

The Sulfiredoxin‐Peroxiredoxin axis promotes urethane‐induced lung adenocarcinoma through the regulation of the tumor microenvironment

Lung adenocarcinoma is the most common type of lung cancer and has a strong association with tobacco smoking. Smoking leads to the accumulation of reactive oxygen species (ROS) and oxidative stress damages to macromolecules, leading to abnormal cell growth & proliferation, transformation, changes in cell metabolism and a variety of other physiological functions. Accumulation of ROS only leads to increased expression of cellular antioxidants that contribute to tumorigenesis and cancer progression. Sulfiredoxin (Srx) is the only enzyme that reduces over‐oxidized forms of typical 2‐Cys Peroxiredoxins (Prxs). Compared to other Prx family members, Srx preferentially interacts with and has a higher binding affinity to Prx4. Our previously work demonstrated that the Srx‐Prx4 axis enhances the RAS/RAF‐MEK‐ERK signaling cascade to promote lung cancer cell proliferation and metastasis. However, the role of the Srx‐Prx axis in de novo lung tumorigenesis has not been explored.In this study, Prx4‐/‐ mice and Srx‐/‐Prx4‐/‐ mice were generated and mouse lung carcinogenesis was induced by a well‐established urethane protocol. Briefly, wildtype and knockout mice at 8‐week of age were injected intraperitoneally with 1g/kg of urethane once per week for three successive weeks. Ten weeks after the administration of urethane, all mice were euthanized and examined for tumors. Our results indicate that both knockout mice have significantly reduced rates on tumor incidence, multiplicity, and size. Compared with tumors from wildtype mice, the rate of cell proliferation in tumors from either knockout mice is significantly decreased.Macrophages are the most abundant immune cells identified in the tumor microenvironment of solid tumors and their presence correlates with reduced survival in most cancers. In addition to all other markers, we also compared the presence of two distinct states of polarized macrophages including the classically activated macrophage (M1) and the alternatively activated macrophage subsets (M2) in urethane‐induced tumors using specific surface marker staining. We found that there are significantly increased numbers of M1 cells in tumors of either Prx4‐/‐ or Srx‐/‐Prx4‐/‐ mice compared with those of wildtype mice. In contrast, there are significantly few numbers of M2 cells in tumors from either knockout mice.It has been shown that M1 macrophages have immune‐stimulatory properties through the expression of a series of pro‐inflammatory cytokines, chemokines, and effector molecules, whereas M2 cells express a wide array of anti‐inflammatory molecules. Therefore, the difference in tumor‐associated macrophages caused by the loss of Prx4 or Srx and Prx4 may contribute to the reduced lung tumorigenesis observed in the mouse model. In addition, bioinformatic analysis shows that Prx4 is overexpressed in the majority of cancers, and the increased expression of Srx and Prx4 both show a strong correlation with poor prognosis in lung adenocarcinoma patients.

Characterization of a Novel Oncolytic Virus for Cancer Therapy

BackgroundOncolytic viruses (OVs) that selectively replicate (oncoselectivity) and lyse (oncotoxicity) cancer cells are emerging as promising cancer therapeutics. A range of DNA and RNA OVs are in different stages of human clinical trials. The presence of preexisting immunity against OVs and their potential pathogenic effects are significant limitations. Using animal viruses that are non‐pathogenic to humans is considered an attractive strategy. We identified a naturally occurring animal virus with potent oncolytic ability, hereafter referred to as oncolytic virus 1 (OV1). We tested the oncoselectivity and oncotoxicity of OV1 using a range of human cancer cells, immortalized cells, and normal non‐cancerous cells. We further evaluated the oncolytic properties of OV1 in vivo using the murine triple‐negative breast cancer (TNBC) model, 4T1.2.MethodsHuman renal (769P), mammary (HCC 1500), colorectal (HRT‐18G), lung (A549), and pancreatic (BxPC‐3) cancer cells, and leukemia (THP‐1) cells, human immortalized embryonic kidney cells (293T), and human primary bronchial epithelial cells (PBEC) were infected with OV1 at a range of multiplicity of infection (MOI) from 0.01 to 5. After 72h of infection, MTS assay to determine cell viability and Caspase‐Glo 3/7 assay to quantify apoptosis were performed. OV1 replication in cultured 4T1.2 cells was assessed by infecting the cells at an MOI of 0.1 and quantifying viral nucleic acid in the supernatants at 72h. Female BALB/c mice were injected with 4T1.2 cells into the fourth mammary gland on the left side. At 19 days post‐tumor implantation, 105TCID50 units of OV1 in 50µl was injected half intratumorally and half intraperitoneally. Mice in the mock treatment group received 50µl of vehicle control. Tumor sizes were measured regularly and at day 35, mice were sacrificed and the number of live cells in the tumor were quantified by flow cytometry.ResultsThere was a significant decrease in the viability of OV1 infected 769P cells (MOI ≥ 0.1), A549 (MOI ≥ 1), THP‐1 (MOI ≥ 2.5), and HCC 1500 (MOI ≥ 2.5) at 72h compared with controls. Notably, there was no significant change in the viability of primary cells (PBEC) and immortalized 293T infected with OV1 at MOI of 0.1 to 2.5 compared to controls. There was a significant increase in caspase activity in 769P cells (at MOI ≥ 0.1), A549 (at MOI ≥ 1), THP‐1 (at MOI ≥ 1), BxPC3 (at MOI ≥ 2.5), HRT‐18G (at MOI ≥ 2.5) and HCC 1500 (at MOI ≥ 0.1). There was no increase in the caspase activity of PBEC at all the tested MOI. OV1 viral replication was observed in 4T1.2 cells in vitro, as evidenced by increasing viral nucleic acid in culture supernatants. BALB/c mice injected with OV1 showed a significant decrease in tumor size and a decrease in the live cell numbers in the tumor compared with mock treated mice.Conclusions and future directionsThe results established the oncoselectivity and oncotoxicity of OV1 and its oncolytic ability in vivo. Further studies are underway to investigate OV1 replication in cells, immune stimulatory properties of OV1 in vivo, and the generation of a recombinant OV1 with increased oncolytic ability.

PHARMACO-THERAPEUTIC REVIEW OF PIPPALYADI AGADA FROM YOGARATNAKAR

Agadtantra is one of the eight branches of Ayurveda which deals with the study of Visha with respect to its types, signs & symptoms, treatment as well as various Agada Kalpas (Medicinal preparations for Visha). Pippalyadi Agada is one of them explained by Yogaratnakara. It is the combination of herbs like, Pippali (Piper longum), Dhanyaka (Coriandrum sativum), Jatamansi (Nardostachys Jatamansi), Lodhra (Symplocos racemosa), Sukshmaela (Elleteria cardimomum), Bruhadela (Amomum subulatum), Maricha (Piper nigrum), Balakam (Valeriana Wallichii) & minerals like, Suwarchika (Salt petre /Potassium nitrate) & Suvarna Gairika (Red oxide of iron) mixed with Honey. It is used in Dushivisha Chikitsa which is a kind of chronic cumulative or residual toxicity. Apart from its anti-toxic nature, ingredients of Pippalyadi Agada have anti-inammatory, anti-oxidant, antifungal, antimicrobial, hepatoprotective and immune-stimulatory properties. So, in this review, pharmacological & therapeutic actions as well as anti-toxic properties of Pippalyadi Agada were discussed.

Molecular Mechanisms of Anti-Neoplastic and Immune Stimulatory Properties of Oncolytic Newcastle Disease Virus.

Oncolytic viruses represent interesting anti-cancer agents with high tumor selectivity and immune stimulatory potential. The present review provides an update of the molecular mechanisms of the anti-neoplastic and immune stimulatory properties of the avian paramyxovirus, Newcastle Disease Virus (NDV). The anti-neoplastic activities of NDV include (i) the endocytic targeting of the GTPase Rac1 in Ras-transformed human tumorigenic cells; (ii) the switch from cellular protein to viral protein synthesis and the induction of autophagy mediated by viral nucleoprotein NP; (iii) the virus replication mediated by viral RNA polymerase (large protein (L), associated with phosphoprotein (P)); (iv) the facilitation of NDV spread in tumors via the membrane budding of the virus progeny with the help of matrix protein (M) and fusion protein (F); and (v) the oncolysis via apoptosis, necroptosis, pyroptosis, or ferroptosis associated with immunogenic cell death. A special property of this oncolytic virus consists of its potential for breaking therapy resistance in human cancer cells. Eight examples of this important property are presented and explained. In healthy human cells, NDV infection activates the RIG-MAVs immune signaling pathway and establishes an anti-viral state based on a strong and uninhibited interferon α,ß response. The review also describes the molecular determinants and mechanisms of the NDV-mediated immune stimulatory effects, in which the viral hemagglutinin-neuraminidase (HN) protein plays a prominent role. The six viral proteins provide oncolytic NDV with a special profile in the treatment of cancer.

PLA/Hydroxyapatite scaffolds exhibit in vitro immunological inertness and promote robust osteogenic differentiation of human mesenchymal stem cells without osteogenic stimuli

Bone defects stand out as one of the greatest challenges of reconstructive surgery. Fused deposition modelling (FDM) allows for the printing of 3D scaffolds tailored to the morphology and size of bone damage in a patient-specific and high-precision manner. However, FDM still suffers from the lack of materials capable of efficiently supporting osteogenesis. In this study, we developed 3D-printed porous scaffolds composed of polylactic acid/hydroxyapatite (PLA/HA) composites with high ceramic contents (above 20%, w/w) by FDM. The mechanical properties of the PLA/HA scaffolds were compatible with those of trabecular bone. In vitro degradation tests revealed that HA can neutralize the acidification effect caused by PLA degradation, while simultaneously releasing calcium and phosphate ions. Importantly, 3D-printed PLA/HA did not induce the upregulation of activation markers nor the expression of inflammatory cytokines in dendritic cells thus exhibiting no immune-stimulatory properties in vitro. Evaluations using human mesenchymal stem cells (MSC) showed that pure PLA scaffolds exerted an osteoconductive effect, whereas PLA/HA scaffolds efficiently induced osteogenic differentiation of MSC even in the absence of any classical osteogenic stimuli. Our findings indicate that 3D-printed PLA scaffolds loaded with high concentrations of HA are most suitable for future applications in bone tissue engineering.

Identification And Characterization Of Phenolic Compounds In Root Extract Of Two Ethnomedicinal Plants Curculigo Orchioides And Asparagus Racemosus

Since early ancient period human being are searching for new drugs with better therapeutic potentials. Traditional medicines are very important because it provide the right direction to the researcher to discover new plant based products against some specific problem and the knowledge has been passed through generation after generation. Curculigo orchioides Gaertn.and Asparagus racemosus Willd. both plants have immune stimulatory properties and are widely used by traditional healers for the treatment of various diseases. The main aim of the present study is to identify the secondary metabolites found in methanolic root extracts of C. orchioides and A. racemosus. Extraction was carried out through standard procedures and the analysis of plant extracts was carried out by using LC-ESI-MS/MS technique in positive and/or negative ionization mode. LC MS study tentatively identified 15 and 19 secondary metabolites from Curculigo orchioides and Asparagus racemosus respectively. Amongst these some were simple phenolic acids such as caffeic acid, quinic acid, p-coumaric acid, sinapic acid, protocatechuic acid, p-hydroxybenzoic acid and vanillic acid. Some others were phenolic acid esters such as chlorogenic acid, di caffeoylquinic acid, p-coumaroyl quinic acid and some flavonoids such as quercetin, rutin, kaempferol, catechin and apigenin were also tentatively identified. Two phenolics caffeic acid and caffeoyl hexoside were further confirmed by MS MS study. This study supports the ethnobotanical claims done by traditional healers of Purulia and Midnapore districts. Characterized phytochemicals were mainly phenolic and flavonoid compounds. Assured levels of phenolics along with other plant constituents in the studied plants supports the ethnobotanical claim done by traditional healers. Further research is needed on structural analysis and bioactivity assay of the identified compounds.

Lactoferrin-Functionalized Noble Metal Nanoparticles as New Antivirals for HSV-2 Infection.

(1) Background: Lactoferrin has been recognized as a potent inhibitor of human herpetic viruses, such as herpes simplex type 1 (HSV-1) and 2 (HSV-2). In this work, we tested if silver and gold nanoparticles modified with lactoferrin (LF-Ag/AuNPs) can become novel microbicides with additional adjuvant properties to treat genital herpes infection. (2) Methods: The antiviral and cytotoxic activities of LF-Ag/AuNPs were tested in human skin HaCaT and vaginal VK-2-E6/E7 keratinocytes. Viral titers and immune responses after treatment with LF-Ag/AuNPs were tested in murine vaginal HSV-2 infection. (3) Results: LF-Ag/AuNPs inhibited attachment and entry of HSV-2 in human keratinocytes much better than lactoferrin. Furthermore, pretreatment with LF-AgNPs led to protection from infection. Infected mice treated intravaginally with LF-Ag/AuNPs showed lower virus titers in the vaginal tissues and spinal cords in comparison to treatment with lactoferrin. Following treatment, vaginal tissues showed a significant increase in CD8+/granzyme B + T cells, NK cells and dendritic cells in comparison to NaCl-treated group. LF-Ag/AuNPs-treated animals also showed significantly better expression of IFN-γ, CXCL9, CXCL10, and IL-1β in the vaginal tissues. (4) Conclusions: Our findings show that LF-Ag/AuNPs could become effective novel antiviral microbicides with immune-stimulant properties to be applied upon the mucosal tissues.

Inhibition of Autophagy by 3-MA Increases Oncolysis Effect of VSV in a Murine Model of Cancer

Vesicular stomatitis virus has been known as a potent antitumor agent because of its selective replication and lysis of tumor cells and immune-stimulating properties. In response to cellular stress and enhanced metabolism, tumor cells activate autophagy, to provide energy for the cells and preventing tumor destruction. Inhibition of autophagy can increase the therapeutic potential of many antitumor methods. This study aimed to check the efficacy of combined VSV and three-methyl adenine (3-MA) in treating a tumor model in mice. TC-1, a line of C57BL/6 mouse lung cells transformed by HPV-16 E7 and E6 oncoproteins, as well as human Ras, were used for experiments. The viability after treatment with the optimized concentration of 3-MA with or without combination with VSV was assessed by MTT. C57BL/6 male mice were injected with TC-1, and after tumor formation, 3-MA and VSV alone or in combination in two different protocols were injected into tumor mice. Tumor size, tumor-specific CTL response, and apoptosis rate were evaluated. The results showed that 3-MA combined with VSV causes more lethality in tumor cells in vitro. In vivo studies also showed that combined VSV and 3-MA treatment inhibits the progression of TC-1 cancer cells with higher efficiency, especially in daily 3-MA treatment along with four doses of VSV injection with four days' intervals. In addition, the rate of apoptosis and cytotoxic T cells activity in the groups injected with 3-MA and the virus were higher than groups receiving each agent alone. In conclusion, the association of VSV with 3-MA increases its oncolysis activity and subsequently more stimulates the immune system against the tumor. This finding suggests a combinational approach for tumor therapy with therapy. Combining oncolytic VSV with 3-MA as an autophagy inhibitor agent can improve the efficacy of tumor treatment. This combination therapy approach enhances apoptosis in tumors as well as T cell cytotoxicity against tumor cells.

A preliminary evaluation of tick cement-cone protein extract for a vaccine against Hyalomma infestation.

Vaccines have been widely exploited to prevent tick-borne infections in cattle. Most vaccines have faced failure in the field because of inconsistency in an immune response. It is presumed that the cement-cone proteins of ticks that participate in the acquisition of blood meal for ticks possess strong immune-stimulating properties and, hence, could be a useful candidate in vaccine development. We evaluated cement-cone proteins of tick Hyalomma anatolicum as a vaccine candidate against infestations of H. anatolicum and H. aegyptium in cattle. The cement-cone proteins were extracted from H. anatolicum to develop stage-reactive and immunogenic cross-reactive vaccine against the infestation of two species of ticks H. anatolicum and H. aegyptium. The immune response of the vaccine was tested against cement-cone proteins starved, partially fed, and richly fed ticks. The findings of the present study demonstrated the cross-reactivity among the two species of ticks that belonged to the same genus (Hyalomma). The antigenic similarity between the two ticks species suggests that a common antigen may possibly be suitable for a vaccine against the two different species of ticks. The results have also indicated that the 23 kDa cement-cone protein of H. anatolicum and H. aegyptium may be responsible for the induction, or elicitation of immunogenic, common stage reactive, and cross-reactive host immune responses with consistent intensity throughout the life stages of ticks. The vaccine based upon cement-cone proteins of ticks may be a useful deterrent against tick-borne infections in cattle in countries like Pakistan.

Accelerated blood clearance and hypersensitivity by PEGylated liposomes containing TLR agonists

Systemic administration of toll-like receptor (TLR) agonists have demonstrated impressive preclinical results as an anti-cancer therapy due to their potent innate immune-stimulatory properties. The clinical advancement has, however, been hindered by severe adverse effects due to systemic activation of the immune system. Liposomal drug delivery systems may modify biodistribution, cellular uptake, and extend blood circulation, and thus, potentially enable systemic administration of TLR agonists at therapeutic doses.In this study, we investigated potential barriers for the administration of TLR agonists formulated in polyethylene glycosylated (PEGylated) liposomes with regards to liposome formulation, TLR agonist, administration route, administration schedule, biodistribution, blood clearance, and anti-PEG antibodies. We found that administration of TLR agonists formulated in PEGylated liposomes led to high anti-PEG antibody titers, which upon multiple intravenous administrations, resulted in accelerated blood clearance and acute hypersensitivity reactions. The latter was found to be associated with anti-PEG IgG antibody and not anti-PEG IgM antibody opsonization. This study highlights the need to carefully design and evaluate nanoparticle delivery systems for immunotherapy as anti-nanoparticle immune responses may challenge the therapeutic application.

EXTH-81. STING ACTIVATION PROMOTES ROBUST IMMUNE RESPONSE AND TUMOR REGRESSION IN GLIOBLASTOMA MODELS

Abstract The immunosuppressive tumor microenvironment in glioblastoma presents a major barrier to effective application of immunotherapeutic approaches in this disease. Although immunotherapy has had a tremendous impact on cancer treatment in the past decade, with hitherto unseen responses at advanced and metastatic stages of the disease glioblastoma remains largely refractory to current immunotherapeutic approaches. The cGAS-STING cytoplasmic double stranded DNA sensing pathway has emerged as a next-generation immunotherapy target with potent local immune stimulatory properties, but has not been well investigated in glioblastoma to date. Here we investigated the presence of STING pathway components in glioblastoma patient specimens and cell lines by Western blotting and immunostaining. The functionality of the pathway was determined by ELISA, and immune infiltrates, and animal survival were investigated in mouse glioblastoma models after treatment with the STING agonist ADU-S100. Our data reveal the presence of STING in human GBM specimens, where it stains strongly in the tumor vasculature, and appears to be activated as evidenced by elevated phosphoTBK1 staining. We show that human GBM cells do not respond agonists, but that GBM explants can respond to STING agonist treatment by secretion of inflammatory cytokines. In murine GBM models, we show a profound shift in the tumor immune landscape after STING agonist treatment, with massive infiltration of the tumor-bearing hemisphere with innate immune cells including inflammatory macrophages, neutrophils and NK populations. Treatment of established murine intracranial GL261 and CT-2A tumors by biodegradable ADU-S100-loaded intracranial implants demonstrated a significant increase in survival in both models and long-term survival with immune memory in GL261. This study reveals therapeutic potential and deep remodeling of the TME by STING activation in GBM and warrants the further examination of STING agonists alone or in combination with other immunotherapies such as cancer vaccines, CAR T cells, or immune checkpoint blockade.

Effects of dietary quercetin on the innate immune response and resistance to white spot syndrome virus in Procambarusclarkii

In recent years, the use of natural products with immune-stimulating and antimicrobial properties has attracted increasing attention in aquaculture researches. In our study, the effect of diet supplemented with quercetin, a flavonoid commonly found in some types of plants substance on the innate immune response and disease resistance in crayfish (Procambarus clarkii) against white spot syndrome virus (WSSV) is reported. It was found that dietary 40 mg/kg quercetin significantly reduced the mortality of crayfish and WSSV copy number after WSSV challenge. Dietary quercetin increased catalase (CAT), and lysozyme (LZM) activity in crayfish. Dietary quercetin increased the expression of NF-κB, anti-lipopolysaccharide factor (ALF) and toll-like receptor (TLR) genes in crayfish. The apoptosis rate of hemocyte was increased by quercetin supplement in crayfish. Our results suggest that dietary quercetin may affect the innate immunity of crayfish and protect crayfish from WSSV infection.

Ameliorative activity of Moringa oleifera on blood parameters of commercial broilers – Mini Review

Moringa is a fast growing plant rich in protein, carbohydrate and mineral contents. This plant is renowned for its antioxidant, antibacterial, antifungal, antiulcer, non tumorous and immune stimulant properties. Leaf powder and aqueous extract can be

Strategies for controlling the innate immune activity of conventional and self-amplifying mRNA therapeutics: Getting the message across.

The recent approval of messenger RNA (mRNA)-based vaccines to combat the SARS-CoV-2 pandemic highlights the potential of both conventional mRNA and self-amplifying mRNA (saRNA) as a flexible immunotherapy platform to treat infectious diseases. Besides the antigen it encodes, mRNA itself has an immune-stimulating activity that can contribute to vaccine efficacy. This self-adjuvant effect, however, will interfere with mRNA translation and may influence the desired therapeutic outcome. To further exploit its potential as a versatile therapeutic platform, it will be crucial to control mRNA's innate immune-stimulating properties. In this regard, we describe the mechanisms behind the innate immune recognition of mRNA and provide an extensive overview of strategies to control its innate immune-stimulating activity. These strategies range from modifications to the mRNA backbone itself, optimization of production and purification processes to the combination with innate immune inhibitors. Furthermore, we discuss the delicate balance of the self-adjuvant effect in mRNA vaccination strategies, which can be both beneficial and detrimental to the therapeutic outcome.