Immunostimulatory Drug Research Articles

Dual immunostimulatory CD73 antibody-polymeric cytotoxic drug complex for triple negative breast cancer therapy

Treatment of triple-negative breast cancer (TNBC) poses significant challenges due to its propensity for metastasis. A key impediment lies in the suppressive immune microenvironment, which fosters tumor progression. This study introduces an approach employing a dual immune-stimulatory CD73 antibody-polymeric cytotoxic drug complex (αCD73-PLG-MMAE). This complex is designed for targeted eradication of TNBC while modulating tumor immunity through mechanisms such as immunogenic cell death (ICD) and interference with the adenosine signaling pathway. By enhancing antitumor immune responses, this strategy offers a highly effective means of treating TNBC and mitigating metastasis. The complex is synthesized by combining αCD73 with poly(L-glutamic acid) (PLG) grafted Fc binding peptides (Fc-III-4C) and Val-Cit-PAB-monomethyl auristatin E (MMAE), exploiting the affinity between αCD73 and Fc-III-4C. αCD73 selectively targets CD73 molecules on both tumor and immune suppressive cells, thereby inhibiting the adenosine pathway. Meanwhile, Val-Cit-PAB-MMAE, activated by cathepsin B, triggers selective release of MMAE, inducing ICD in tumor cells. In a 4T1 tumor model, αCD73-PLG-MMAE significantly enhances drug accumulation in tumors by 4.13-fold compared to IgG-PLG-MMAE, leading to suppression of tumor growth and metastasis. Furthermore, it synergistically augments the antitumor effects of αPD-1, resulting in a tumor inhibition rate of 92% as compared to 21% with αPD-1 alone. This study thus presents a pioneering therapeutic strategy for TNBC, emphasizing the potential of targeted immunomodulation in cancer treatment. Statement of SignificanceAntibody-drug conjugate (ADC) therapy holds promise for treating triple-negative breast cancer (TNBC). However, the current ADC, sacituzumab govitecan, fails to overcome the crucial role of adenosine in the suppressive immune microenvironment characteristic of this "cold tumor". Here, we present a dual immune-stimulatory complex, αCD73-PLG-MMAE, which targets TNBC specifically and modulates tumor immunity through mechanisms such as immunogenic cell death (ICD) and interference with the adenosine signaling pathway. Thus, it kills tumor cells with cytotoxic drugs, comprehensively regulates immunosuppression, and restores a durable immune response. This study proposes an antibody-polymeric drug complex with immunomodulatory and immunoagonist roles, offering new insights into TNBC treatment.

#1815 The neutrophil/lymphocyte, neutrophil/HDL-cholesterol, and monocyte/HDL-cholesterol ratios as simple markers of low-grade inflammation in CKD patients

Abstract Background and Aims Chronic kidney disease (CKD) has been associated with inflammation and oxidative stress, as well as with high cardiovascular (CV) morbidity and mortality. The neutrophil/lymphocyte (NLR), neutrophil/HDL-cholesterol (NHR), and monocyte/HDL-cholesterol (MHR) ratios have been linked both to inflammation and increased CV risk in a variety of diseases. The aim of this study was to investigate changes in these ratios in CKD patients of stage 2-5 without any clinical and laboratory signs of inflammation, and to compare these indicators with those in healthy volunteers. Method A retrospective cross-sectional study was conducted in 88 healthy volunteers and 187 CKD patients (50 of stage 2, 44 of stage 3a, 45 of stage 3b and 48 of stage 4-5). Patients with acute renal impairment, nephrotic proteinuria, infections, systemic inflammation, malignancies and hematologic diseases were excluded, as were those receiving hypolipidemic, corticosteroid, immunosuppressive or immunostimulant drugs. The whole blood samples were collected and the absolute numbers of neutrophils, lymphocytes, and monocytes were used for calculation of NLR, NHR and MHR, respectively. Serum creatinine, HDL-cholesterol, C-reactive protein (CRP) and albumin were measured and eGFR calculated according to the CKD EPI equation. Statistical analysis was performed with the Mann-Whitney, Shapiro–Wilk, Kruskal-Wallis, and Spearman's correlation tests using the SPSS software. Results The NLR, NHR and MHR were higher in CKD patients when compared to healthy volunteers (P < 0,001), but only NLR and NHR increased significantly across CKD stages (P < 0,01). The ratio differences between CKD stages were more evident for NHR which was also in best correlation with eGFR (ρ = −0,406, P < 0,001). The CRP values increased across CKD stages as well (P < 0,01) although remaining within the reference range. The NLR, NHR and MHR were significantly related to CRP with NHR providing the best correlation (ρ = 0,289, P < 0,001). Conclusion This study showed that NLR and NHR increased across CKD stages 2-5 and could be used to identify low-grade inflammation in this population even within normal CRP values.

Host-functionalization of macrin nanoparticles to enable drug loading and control tumor-associated macrophage phenotype.

Macrophages are critical regulators of the tumor microenvironment and often present an immuno-suppressive phenotype, supporting tumor growth and immune evasion. Promoting a robust pro-inflammatory macrophage phenotype has emerged as a therapeutic modality that supports tumor clearance, including through synergy with immune checkpoint therapies. Polyglucose nanoparticles (macrins), which possess high macrophage affinity, are useful vehicles for delivering drugs to macrophages, potentially altering their phenotype. Here, we examine the potential of functionalized macrins, synthesized by crosslinking carboxymethyl dextran with L-lysine, as effective carriers of immuno-stimulatory drugs to tumor-associated macrophages (TAMs). Azide groups incorporated during particle synthesis provided a handle for click-coupling of propargyl-modified β-cyclodextrin to macrins under mild conditions. Fluorescence-based competitive binding assays revealed the ability of β-cyclodextrin to non-covalently bind to hydrophobic immuno-stimulatory drug candidates (Keq ~ 103 M-1), enabling drug loading within nanoparticles. Furthermore, transcriptional profiles of macrophages indicated robust pro-inflammatory reprogramming (elevated Nos2 and Il12; suppressed Arg1 and Mrc1 expression levels) for a subset of these immuno-stimulatory agents (UNC2025 and R848). Loading of R848 into the modified macrins improved the drug's effect on primary murine macrophages by three-fold in vitro. Intravital microscopy in IL-12-eYFP reporter mice (24h post-injection) revealed a two-fold enhancement in mean YFP fluorescence intensity in macrophages targeted with R848-loaded macrins, relative to vehicle controls, validating the desired pro-inflammatory reprogramming of TAMs in vivo by cell-targeted drug delivery. Finally, in an intradermal MC38 tumor model, cyclodextrin-modified macrin NPs loaded with immunostimulatory drugs significantly reduced tumor growth. Therefore, efficient and effective repolarization of tumor-associated macrophages to an M1-like phenotype-via drug-loaded macrins-inhibits tumor growth and may be useful as an adjuvant to existing immune checkpoint therapies.

Investigation on the Efficacy of Two Food Supplements Containing a Fixed Combination of Selected Probiotics and β-Glucans or Elderberry Extract for the Immune System: Modulation on Cytokines Expression in Human THP-1 and PBMC

Several herbal and other natural products are used as ingredients in food supplements to strengthen immunity even if, very often, marketed products are proposed without a clear rationale or experimental evidence. In this study, we aimed to investigate the effect on human monocytes (THP-1) and on ex vivo human peripheral blood mononuclear cells (PBMC) of two formulations, one containing Bifidobacterium animalis subsp. lactis Bl-04® with β-glucans (for adults) and one containing Lactobacillus rhamnosus CRL1505 with elderberry extract (for children). We compared formulations with single ingredients, with bacterial lipopolysaccharide (LPS) and the drug pidotimod; cytokines expression level was evaluated testing different concentrations of samples at two exposure times. As expected, LPS caused a non-specific huge upregulation of cytokines expression both in THP-1 and in PBMC, whereas pidotimod mainly upregulated IL-2 in PBMC and IL-8 in THP-1. The two formulations showed a difference between a pro-inflammatory stimulus such as LPS, and also from an immunostimulant drug, such as pidotimod, as they mainly upregulated the expression of IL-6 and IL-10 in PBMC but not in THP-1, in a concentration-dependent mode. Probiotics were shown to play a major role, but β-glucans and elderberry extract exerted a synergistic activity. This work demonstrated that combining selected probiotics with other natural products having immunomodulatory properties is an interesting strategy to develop innovative formulations in the sector of food supplements.

The potential immunomodulatory effect of levamisole in humans and farm animals.

This study conducted a literature review to investigate the immunomodulatory effect of levamisole in both humans and farm animals. The following procedure was followed for database searching: PubMed, Google Scholar, Web of Science, and Cochrane Library. All research works were updated to September 2022. The terms used in the literature search were included: ("levamisole" OR "immunity" OR "immune system") AND ("adjuvant" OR "fish" OR "poultry" OR "farm animal" OR "cattle" OR "sheep"). The current review enlightens the extensive potential of levamisole as an adjuvant immunotherapeutic agent and explains its divergent applications beyond its antiparasitic use as an adjuvant, dietary supplement, immunostimulant, antiviral, and anti-cancer drug in humans and farm animals. In the articles examined, various mechanisms have been proposed for levamisole immunoprotective effects, but hormonal alteration and stress hormone reduction are indicated as the main mechanisms in various animal species.

Micronanoparticled risedronate exhibits potent vaccine adjuvant effects

Micro/Nano-scale particles are widely used as vaccine adjuvants to enhance immune response and improve antigen stability. While aluminum salt is one of the most common adjuvants approved for human use, its immunostimulatory capacity is suboptimal. In this study, we modified risedronate, an immunostimulant and anti-osteoporotic drug, to create zinc salt particle-based risedronate (Zn-RS), also termed particulate risedronate. Compared to soluble risedronate, micronanoparticled Zn-RS adjuvant demonstrated increased recruitment of innate cells, enhanced antigen uptake locally, and a similar antigen depot effect as aluminum salt. Furthermore, Zn-RS adjuvant directly and quickly stimulated immune cells, accelerated the formulation of germinal centers in lymph nodes, and facilitated the rapid production of antibodies. Importantly, Zn-RS adjuvant exhibited superior performance in both young and aged mice, effectively protecting against respiratory diseases such as SARS-CoV-2 challenge. Consequently, particulate risedronate showed great potential as an immune-enhancing vaccine adjuvant, particularly beneficial for vaccines targeting the susceptible elderly.

Levamisole incorporation in fish feed – Ensuring the medication dose and avoidance of leaching into the water

Levamisole, an anthelmintic and immunostimulant drug, has been studied as a promising alternative for aquaculture use. While oral administration through feeding is the main route of administration in fish farming, no studies evaluating methods of levamisole incorporation into the feed have been reported so far. Therefore, this study aimed to evaluate potential procedures for levamisole incorporation in extruded fish feed using ethyl cellulose, gelatin, or vegetable oil, to avoid drug leaching to the water during the animal’s medication. A suitable LC-MS/MS method was optimized (full factorial design), validated, and applied to evaluate the efficiency of the process, the homogeneity of the drug concentration, and the leaching rate. The method has been demonstrated to be selective, precise (RSD < 4.9%), accurate (recovery > 98.4%), and linear (r > 0.99, 125–750 mg kg−1). The incorporation procedures using the three coating agents showed high incorporation efficiency (70%) and a homogeneous drug concentration among the extruded feed pellets. A low levamisole leaching rate was verified in the feed prepared using the ethyl cellulose coating procedure (4.3% after 15 min of immersion in the water). On the other hand, fish feed coated with gelatin and oil resulted in a high leaching rate (30–35% after 15 min). Thus, this study shows that coating ethyl cellulose may be a promising procedure for levamisole incorporation in fish feed and with the potential to enhance its use in animal production while reducing environmental contamination.

Is imiquimod a promising drug to treat oral mucosa diseases? A scoping review and new perspectives.

Imiquimod (IMQ) is a chemotherapeutic and immunostimulant drug that is applied topically, demonstrating antitumor and antiviral activities. The objective of this review was to compile data on the off-label use of IMQ in oral mucosal diseases. IMQ has exhibited effectiveness in the treatment of various oral mucosal conditions, including oral carcinogenic lesions, neoplasms, HPV-related lesions and autoimmune disorders. Although IMQ holds promise as a potential strategy for addressing oral mucosal lesions, it is important to note that significant side effects have been frequently reported. Nonetheless, it is crucial to develop and test new technological systems, such as the combination of nanotechnology with innovative drug delivery platforms. These advancements aim to minimize side effects and prolong the drug's contact time with the mucosa, preventing its removal by salivary flow.

Blockade of IL-10 Signaling Ensures Mifamurtide Efficacy in Metastatic Osteosarcoma.

Osteosarcoma (OS) is the most common primary malignancy of the bone, highly aggressive and metastasizing, and it mainly affects children and adolescents. The current standard of care for OS is a combination of surgery and chemotherapy. However, these treatment options are not always successful, especially in cases of metastatic or recurrent osteosarcomas. For this reason, research into new therapeutic strategies is currently underway, and immunotherapies have received considerable attention. Mifamurtide stands out among the most studied immunostimulant drugs; nevertheless, there are very conflicting opinions on its therapeutic efficacy. Here, we aimed to investigate mifamurtide efficacy through in vitro and in vivo experiments. Our results led us to identify a new possible target useful to improve mifamurtide effectiveness on metastatic OS: the cytokine interleukin-10 (IL-10). We provide experimental evidence that the synergic use of an anti-IL-10 antibody in combination with mifamurtide causes a significantly increased mortality rate in highest-grade OS cells and lower metastasis in an in vivo model compared with mifamurtide alone. Overall, our data suggest that mifamurtide in combination with an anti-IL-10 antibody could be proposed as a new treatment protocol to be studied to improve the outcomes of OS patients.

Secretory IgA in patients with COVID-19 at different regimens of using multicomponent vaccine Immunovac-VP-4

The mucosal immunity performs an important function in prevention of respiratory infections including COVID-19. The search for approaches to activate the synthesis of post-infectious antibodies by correcting the factors of innate and adaptive immunity at mucous membranes of respiratory tract in patients with infection caused by the new coronavirus may be relevant for the treatment of patients with COVID-19. The aim of our study was to assess the concentrations of sIgA in the upper respiratory tract in patients with a confirmed diagnosis of Coronavirus infection caused by the COVID-19, and to evaluate the effect of an immunostimulating drug of bacterial origin upon the sIgA secretion.&#x0D; The patients were divided into two groups: group 1 (n = 45), received basic therapy; group 2 (n = 33), in addition to basic therapy, received the bacterial vaccine Immunovac-VP-4 according to a combined scheme. The biomaterial sampling was carried out by scraping of epithelial cells from the nasal mucosa, pharyngeal scraping and salivary gland secretion on days 1, 14 and 30 of the study. sIgA levels in all biological fluids were studied using ELISA technique (JSC Vector-Best, Russia).&#x0D; 14 days after the start of observation, the dynamics of sIgA levels in nasal scrapings in group 1showed a significant decrease relative to the baseline values (p = 0.02), whereas the level of sIgA remained unchanged during the specified period (p = 0.07) in the group of patients receiving, along with basic therapy, additional Immunovac-VP-4 treatment. The dynamics of sIgA level in pharyngeal scrapings in the group of patients receiving only basic therapy did not change throughout the study period. Menwhile, the group of patients receiving basic therapy supplemented with Immunovac-VP-4 showed a significant increase in sIgA levels by the 30th day of follow-up over the baseline values (p = 0.02). The level of sIgA in salivary gland secretions did not differ significantly between the study groups during the entire follow-up period.&#x0D; The results of our study showed that, in order to assess the state of mucosal immunity in patients with COVID-19, one may recommend determination of sIgA in nasal secretions. The Immunovac-VP-4 prescribed in complex therapy is accompanied by an increase in the sIgA levels at the mucous surfaces of the respiratory tract.

Effectiveness of the Study of Plant-Contaınıng Immunostımulants in Poultry Industry

The effect of the plant-derived preparation (Tilia cordata Mill., Thymus kotschyanus Boiss and Hohen., Matricaria chamomilla L.) Imuneo on the immunity indices in chickens vaccinated against avian infectious bronchitis was studied, and the immunostimulatory efficacy was evaluated. In the group of chickens treated with immunostimulants, a significant increase in their immune status was recorded. The immunostimulating effect of Imuneon was determined. Microbicidal and immunostimulating properties of plant-derived Hedalin preparation were studied. The expediency of using immunostimulating drugs in poultry complexes is presented.

Antiviral activity of basidial fungus Inonotus obliquus aqueous extract against SARS-CоV-2 virus (Coronaviridae: Betacoronavirus: Sarbecovirus) in vivo in BALB/c mice model

The COVID-19 pandemic combined with seasonal epidemics of respiratory viral diseases requires targeted antiviral prophylaxis with restorative and immunostimulant drugs. The compounds of natural origin are low-toxic, but active against several viruses at the same time. One of the most famous compounds is Inonotus obliquus aqueous extract. The fruit body of basidial fungus I. obliquus is called Chaga mushroom. The aim of the work ‒ was to study the antiviral activity of I. obliquus aqueous extract against the SARS-CoV-2 virus in vivo. Antiviral activity of I. obliquus aqueous extract sample (#20-17) was analyzed against strain of SARS-CoV-2 Omicron ВА.5.2 virus. The experiments were carried out in BALB/c inbred mice. The SARS-CoV-2 viral load was measured using quantitative real-time PCR combined with reverse transcription. The severity of lung tissue damage was assessed by histological methods. The peak values of the viral load in murine lung tissues were determined 72 hours after intranasal inoculation at dose of 2,85 lg TCID50. The quantitative real-time PCR testing has shown a significant decrease in the viral load compared to the control group by 4,65 lg copies/ml and 5,72 lg copies/ml in the lung tissue and nasal cavity samples, respectively. Histological methods revealed that the decrease in the number and frequency of observed pathomorphological changes in murine lung tissues depended on the introduction of the compound under study. The results obtained indicate the possibility of using basidial fungus Inonotus obliquus aqueous extract as a preventive agent against circulating variants of SARS-CoV-2 virus.

КЛИНИКО-ФАРМАКОЛОГИЧЕСКИЕ ОСОБЕННОСТИ ПРИМЕНЕНИЯ ПРЕПАРАТОВ НОРМАЛЬНОГО ИММУНОГЛОБУЛИНА ЧЕЛОВЕКА В ПРАКТИКЕ ВРАЧА

Immunostimulating (immunomodulatory) drugs belong to the group of immunotropic drugs, the therapeutic effect of which is associated with a predominant effect on the human immune system. Medicines based on human normal immunoglobulin are immunostimulatory drugs of natural origin. In the practice of a doctor, immunoglobulins are made from human plasma and are more often used in the form for intravenous and subcutaneous administration. Immunoglobulins differ in the ratio of various subclasses of immunoglobulins G (IgG), as well as immunoglobulin class A (IgA), individual immunoglobulins are enriched with antibodies of the IgM classes, and specific immunoglobulins contain a significantly larger amount of IgG class antibodies against certain pathogens than standard immunoglobulins. Approaches to the classification of immunoglobulins, the mechanism of action, indications, contraindications, as well as clinical and pharmacological features of the use of human normal immunoglobulin, drugs available on the market in the Republic of Belarus are described, recommendations are given for their use in clinical practice. The possibilities of using immunoglobulins as a replacement (for immunodeficiency states, agammaglobulinemia) and immunomodulatory (for autoimmune diseases) therapy are presented. The possibility of using intravenous immunoglobulin for the treatment of Kawasaki syndrome, idiopathic thrombocytopenic purpura, Guillain-Barré syndrome, autoimmune polyneuropathy, diseases arising in oncological, surgical and obstetric practice, as well as for complex therapy of sepsis, including in newborns, is considered. Timely appointment of immunoglobulins in optimal doses will reduce the duration of therapy in patients with immunosuppression of various origins, reduce bleeding in immune thrombocytopenia.

Modulation of immunological responses by aqueous extract of Datura stramonium L. seeds on cyclophosphamide-induced immunosuppression in Wistar rats

BackgroundDatura stramonium L. (Solanaceae) is used traditionally in west Africa to treat asthma, epilepsy, rheumatoid arthritis, filariasis microbial infections and conjunctivitis. This study investigated the immunomodulatory effects of aqueous seed extract of D. stramonium L. (ASEDS) on Wistar rats.MethodsThirty Wistar albino rats (180–200 g) were randomized into 6 groups (n = 5). Group 1 received distilled water only. Rats in groups 2–6 were pretreated with 10 mg/kg body weight (b.w.) Cyclophosphamide orally for 27-days to induce immunosuppression. Thereafter, they received treatment orally for 28 days as follows: Group 2 (distilled water), group 3 (5 mg/kg b.w. Levamisole), groups 4–6 (60, 90 and 120 mg/kg b.w. ASEDS, respectively). HPLC was used to determine major compounds in ASEDS. The effects of ASEDS on immune cells, immunoglobulins A, G and M levels, lipoproteins, and antioxidant status of rats were evaluated.ResultsASEDS indicated high content of Acutumine, Quinine, Catechin, Chlorogenic acid, Gallic acid, Quercetin, Vanillic acid, Luteolin, Formosanin C, Saponin, Cyanidin, Tannic acid, 3-Carene, Limonene and α-terpineol. Cyclophosphamide triggered significant (p < 0.05) reduction in total leucocyte count and differentials, IgA, IgG, high-density lipoproteins (HDL), catalase, superoxide dismutase, glutathione peroxidase, vitamins A, C and E levels of untreated rats. Administration of ASEDS led to significant (p < 0.05) improvement in immune cell counts, immunoglobulin synthesis, high-density lipoprotein concentration, and antioxidant status of rats in the treated groups.ConclusionsThe results obtained from the study showed the immunomodulatory activity of ASEDS, thereby indicating its potential in immunostimulatory drug discovery.

КЛИНИЧЕСКАЯ ОЦЕНКА ЭФФЕКТИВНОСТИ ТЕРАПИИ СОПРОВОЖДЕНИЯ ПРИ ОРТОДОНТИЧЕСКОМ ЛЕЧЕНИИ ПОДРОСТКОВ С ХРОНИЧЕСКИМИ ЗАБОЛЕВАНИЯМИ ВЕРХНИХ ОТДЕЛОВ ЖЕЛУДОЧНО-КИШЕЧНОГО ТРАКТА

The aim of the work is to provide a clinical assessment of the efficacy of therapy for accompanying orthodontic treatment of adolescents with chronic diseases of the upper part gastrointestinal tract. Material and methods. There were 109 adolescents at the age of 12–15 years with dental anomalies and gastroduodenal pathology (chronic gastritis, chronic gastroduodenitis, biliary dysfunction) were taken for orthodontic treatment. The state of oral hygiene, periodontal tissues and hard tooth tissues in the dynamics of orthodontic treatment was evaluated. We used the Silness-Loe and Stallard hygiene indexes, periodontal indexes – PMA, stomatorrhagia, Schiller-Pisarev, dental calculus, and calculated the reduction of caries.&#x0D; Results. Phased application of accompanying therapy (a calcium preparation with vitamin D3 and trace elements (Calcemin), magnesium (Magne B6), an immunostimulating drug of bacterial origin (Imudon) for the orthodontic treatment of adolescents with chronic pathology of the digestive tract shows an anti-inflammatory and caries static effect.

Abstract 2007: Use of an anti-IL-10 antibody to improve Mifamurtide efficacy on aggressive osteosarcoma cells

Abstract Osteosarcoma is a primary malignant bone tumor highly aggressive and metastasizing, composed of mesenchymal cells producing osteoid and immature matrix. Osteosarcoma is the most frequent bone tumor and it mainly affects children and adolescents. The main treatment option is a combination of surgery and chemotherapy. Research into new therapeutic strategies to improve the quality of life of patients is currently underway. The development of the anti-tumor immunostimulant drug Mifamurtide is part of this research. There are very conflicting opinions on the therapeutic validity of this drug. Therefore, we tested experimentally the effectiveness of Mifamurtide in three cell lines of osteosarcoma (MG-63, HOS, 143B) with different grades of malignancy, grown in co-culture with macrophage to allow the correct internalization of the drug. Through in vitro experiments, we analyzed the effects of Mifamurtide on tumor cell viability, on cellular signaling pathway and on macrophage polarization. Results show that Mifamurtide loses its effectiveness as the tumor malignity increases. The most aggressive tumor cells are not affected by the therapy, probably because they implement methods of resistance to apoptosis. Furthermore, they would be able to affect the tumor microenvironment to their own advantage. To better understand Mifamurtide impact on tumor microenvironment, we also performed a cytokine assay on the culture medium in which the cells grew. The most significant results were identified in the interleukine-10 (IL-10): IL-10 is expressed in higher quantities in the highest-grade osteosarcoma cells. This anti-inflammatory cytokine can inhibit the synthesis of pro-inflammatory cytokine and suppresses the antigen presentation capacity of APC cells. This allows cancer cells to evade immune surveillance mechanisms. In vivo and in vitro results show that the synergic use of an anti-IL-10 antibody in combination with Mifamurtide can improve its efficacy. Here, we provide experimental evidence that the addition of an anti-IL-10 antibody to Mifamurtide causes a significant death rate in more aggressive osteosarcoma cells and lower metastasis, compared to Mifamurtide only. In conclusion, considering our data, it could be proposed a new treatment protocol for metastatic osteosarcoma that includes the use of an anti-IL10 antibody integrated with the administration of Mifamurtide to increase its effectiveness in this clinical context. Citation Format: Nicoletta Nastasi, Angela Subbiani, Maria Letizia Taddei, Lucia Scala, Angela Tamburini, Claudio Favre, Maura Calvani. Use of an anti-IL-10 antibody to improve Mifamurtide efficacy on aggressive osteosarcoma cells [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 2007.

Immunological substantiation of complex therapy in patients with mild HPV-associated cervical intraepithelial neoplasia

Introduction. As is known, the development of invasive cervical cancer is preceded by cervical intraepithelial neoplasia of varying severity, which is a pathological process in which cells with varying degrees of atypia and impaired differentiation appear in the thickness of the stratified squamous epithelium of the cervix.Objective. To study the effect of complex therapy in patients with mild HPV-associated cervical intraepithelial neoplasia on the dynamics of local cytokine status and autoimmunity parameters.Materials and methods. The study included 86 patients aged 35 to 40 years with morphologically verified mild cervical intraepithelial neoplasia. The main group consisted of 52 patients who, along with radio wave destruction of the cervix, were prescribed an immunostimulating drug with antiviral activity “Groprinosin-Richter” 1000 mg 3 times a day for 10 days 10–14 days before destruction and similar 2 courses after 10–14 days after it. The comparison group included 34 patients who underwent only radio wave destruction of the cervix.Results. An analysis of the outcomes of low-grade squamous intraepithelial lesions after 6 months showed that in the main group, CIN I regression was observed in 92.3% of patients, persistence – in 7.7% of cases, while in the comparison group, CIN I regression was noted in 73,5% of cases, persistence – in 26.5% of patients.Conclusions. Conducting complex therapy, including radio wave destruction and the use of a drug with immunostimulating and antiviral activity “Groprinosin-Richter” in patients with low-grade HPV-associated cervical intraepithelial neoplasia, leads to the elimination of the initial imbalance of cytokines and normalization of autoantibody levels, helping to reduce the likelihood of HPV persistence and CIN progression to invasive cervical cancer uterus.

Inactivated Cowpea Mosaic Virus in Combination with OX40 Agonist Primes Potent Antitumor Immunity in a Bilateral Melanoma Mouse Model.

Viral immunotherapies are being recognized in cancer treatment, with several currently approved or undergoing clinical testing. While contemporary approaches have focused on oncolytic viral therapies, our efforts center on the development of plant virus-based cancer immunotherapies. In a previous work, we demonstrated the potent efficacy of the cowpea mosaic virus (CPMV), a plant virus that does not replicate in animals, applied as an in situ vaccine. CPMV is an immunostimulatory drug candidate, and intratumoral administration remodels the tumor microenvironment leading to activation of local and systemic antitumor immunity. Efficacy has been demonstrated in multiple tumor mouse models and canine cancer patients. As wild-type CPMV is infectious toward various legumes and because shedding of infectious virus from patients may be an agricultural concern, we developed UV-inactivated CPMV (termed inCPMV) which is not infectious toward plants. We report that as a monotherapy, wild-type CPMV outperforms inCPMV in mouse models of dermal melanoma or disseminated colon cancer. Efficacy of inCPMV is less than that of CPMV and similar to that of RNA-free CPMV. Immunological investigation using knockout mice shows that inCPMV does not signal through TLR7 (toll-like receptor); structure-function studies indicate that the RNA is highly cross-linked and therefore unable to activate TLR7. Wild-type CPMV signals through TLR2, -4, and -7, whereas inCPMV more closely resembles RNA-free CPMV which signals through TLR2 and -4 only. The structural features of inCPMV explain the increased potency of wild-type CPMV through the triple pronged TLR activation. Strikingly, when inCPMV is used in combination with an anti-OX40 agonist antibody (administered systemically), exceptional efficacy was demonstrated in a bilateral B16F10 dermal melanoma model. Combination therapy, with in situ vaccination applied only into the primary tumor, controlled the progression of the secondary, untreated tumors, with 10 out of 14 animals surviving for at least 100 days post tumor challenge without development of recurrence or metastatic disease. This study highlights the potential of inCPMV as an in situ vaccine candidate and demonstrates the power of combined immunotherapy approaches. Strategic immunocombination therapies are the formula for success, and the combination of in situ vaccination strategies along with therapeutic antibodies targeting the cancer immunity cycle is a particularly powerful approach.

ПРОТИВОВИРУСНЫЕ ПРЕПАРАТЫ И ИХ ЭФФЕКТИВНОСТЬ ПРИ ЛЕЧЕНИИ КАЛЬЦИВИРОЗА КОШЕК

Calicivirus disease of cats is widespread in the world. Members of the Felidae family are the only hosts susceptible to FCV infection. The prevalence of FCV is directly related to the number of cats present in the kennel, and is higher in large groups, for example, in colonies and shelters, where it can reach from 25 to 40%. Currently, it is relevant to study the frequency of manifestation and distribution areas of this animal disease, the search for antiviral drugs that are effective against the causative agent of calicivirus infection in cats to optimize therapeutic measures. The virus is able to cause lesions on its own, without the presence of bacteria, and a secondary bacterial infection enhances the effect of the virus, promotes its penetration into deeper parts of the respiratory tract, leading to the development of pneumonia, or chronic rhinitis, sinusitis or conjunctivitis. The development of effective methods for the treatment of calicivirus infection in cats is one of the promising areas of veterinary science and practice. The article presents the results of studies obtained in assessing the effectiveness of the use of antiviral and immunostimulating drugs from various manufacturers: "Fosprenil" (CJSC "Micro-plus", Russia), "Maksidin" (CJSC "Micro-plus", Russia), "Feliferon" (BIOINVEST, Russia). The use of the proposed immunomodulatory and antiviral drugs in the scheme of complex therapy of calicivirus infection in cats had a positive effect on the activation of certain stages of immune protection. Based on the conducted studies, it was found that the most effective method of treating calicivirosis in cats is the use of the drug "Feliferon" intramuscularly 200,000 IU, 1 time a day, 5 days in the complex therapeutic scheme.

Properties of the drug Trekrezan to modulate protein synthesis in cells

New factors contributing to the processes of hormesis involving the peroxisome activator receptor coactivator 1 alpha gene (PGC-1α) and linking the mechanisms of energy regulation with the functions of protein synthesis were studied. Coordinated stimulation of the expression of the tryptophanyl-tRNA synthetase gene, its activity in the synthesis of the tertiary messenger aP3A and the expression of the PGC-1α gene in in vivo and in vitro systems, under the action of the immunostimulating and adaptogenic drug Trekrezan, was shown. It has been suggested that PGC—1α is a promising target for the therapeutic effect of drugs stimulating gene activity.