Rapamycin delays growth of Wnt-1 tumors in spite of suppression of host immunity

<p>Abstract</p> <p>Background</p> <p>Rapamycin, an inhibitor of mammalian target of Rapamycin (mTOR), is an immunosuppressive agent that has anti-proliferative effects on some tumors. However, the role of Rapamycin-induced immune suppression on tumor progression has not been examined.</p> <p>Methods</p> <p>We developed a transplantation model for generation of mammary tumors in syngeneic recipients that can be used to address the role of the immune system on tumor progression. We examined the effect of Rapamycin on the immune system and growth of MMTV-driven Wnt-1 mammary tumors which were transplanted into irradiated and bone marrow-reconstituted, or naïve mice.</p> <p>Results</p> <p>Rapamycin induced severe immunosuppression and significantly delayed the growth of Wnt-1 tumors. T cell depletion in spleen and thymus and reduction in T cell cytokine secretion were evident within 7 days of therapy. By day 20, splenic but not thymic T cell counts, and cytokine secretion recovered. We determined whether adoptive T cell therapy enhances the anti-cancer effect using <it>ex vivo </it>generated Rapamycin-resistant T cells. However, T cell transfer during Rapamycin therapy did not improve the outcome relative to drug therapy alone. Thus, we could not confirm that suppression of T cell immunity contributes to tumor growth in this model. Consistent with suppression of the mTOR pathway, decreased 4E-BP1, p70 S6-kinase, and S6 protein phosphorylation correlated with a decrease in Wnt-1 tumor cell proliferation.</p> <p>Conclusion</p> <p>Rapamycin has a direct anti-tumor effect on Wnt-1 breast cancer <it>in vivo </it>that involves inhibition of the mTOR pathway at doses that also suppress host immune responses.</p

Biomaterials based on chitosan and its derivatives to prevent adhesion formation

An increase in the number and volume of surgical interventions leads to an increase in the frequency of postoperative adhesions. The development of the adhesion process in the abdominal cavity causes pain, a decrease in the quality of life of patients, a violation of the reproductive function of women as well as acute adhesion intestinal obstruction. Recently, polymer biomaterials, including those based on chitosan, have been widely used for the prevention of adhesions. Due to their biocompatibility and biodegradation ability, they do not require repeated operations to extract the material. It is believed that these materials act as barriers, physically separating the damaged surfaces. The molecular mechanism of their action is still poorly understood. In this review, the main mechanisms of adhesion formation, as well as ways to prevent them with the help of materials based on chitosan and its derivatives, are discusse

The role of complement in the susceptibility to invasive aspergillosis

National audienc

Innate immunity and the role of epithelial barrier during aspergillus fumigatus infection

Fungi are the most important eukaryotic infective agents in Europe which largely overpass parasite infections. Total number of people dying of fungal infection is increasing and this trend is likely to continue due to the increase in immunosuppressive treatments. The opportunistic pathogen Aspergillus fumigatus (Af) is a saprophytic filamentous fungus that can cause invasive pulmonary diseases in immuno-compromised hosts. In veterinary medicine aspergillosis is also a recurrent problem since it infects various species, birds are particularly susceptible. It propagates through airborne conidia (spores), which are inhaled into the small airways where they may germinate and initiate an infection. The host epithelium has permanent contact with the environment and a multitude of diverse microorganisms, resulting in a network of the host’s defense mechanisms. Pathogens use various strategies to invade epithelial barriers, to exploit eukaryotic host function to their own benefit and disseminate throughout the host using the epithelium as a reservoir. The current revue will discuss the ways how epithelial and innate immunity cells can contlol Af infection. We will focus on Af strategies for the host’s invasion, antifungal innate immune response and antimicrobial activities of the respiratory epithelial cells

Encapsulation of Allergens into Core–Shell Chitosan Microparticles for Allergen-Specific Subcutaneous Immunotherapy

IgE-mediated allergic reaction occurs in response to harmless environmental compounds, such as tree and grass pollen, fragments of household microorganisms, etc. To date, the only way to treat IgE-mediated allergy is allergen-specific immunotherapy (ASIT), which consists of a prolonged subcutaneous administration of allergen extracts or recombinant proteins. The long duration of the treatment, the cost and the risk of life-threatening adverse reactions are the main limiting factors for ASIT. The aim of this work was to develop allergen proteins encapsulated in chitosan-based microparticles that can be safely administered at high doses and in a rash protocol. The egg white allergen, Gal d 1 protein, was used as a model antigen. The protein was packed into core–shell type microparticles (MPs), in which the core was formed with succinyl chitosan conjugated to Gal d 1, subsequently coated with a shell formed by quaternized chitosan. The obtained core–shell MPs containing Gal d 1 in the core (Gal-MPs) were non-toxic to macrophage and fibroblast cell lines. At the same time, Gal-MPs were quickly engulfed by bone marrow-derived dendritic cells or RAW264.7 macrophage cells, as was visualized using flow cytometry and confocal microscopy. Encapsulated Gal d 1 was not recognized by Gal d 1-specific IgE in ELISA. Female BALB/c mice were immunized with Gal-MPs subcutaneously three times a week for 2 weeks. Immunization of mice resulted in IgG titers 1250 ± 200 without IgE production. Allergy in control and vaccinated mice was induced by low-dose Gal d 1 injections in the withers of mice. IgE was induced in control-sensitized but not in the vaccinated mice. Thus, preventive vaccination with the encapsulated allergens is safe and rapid; it significantly reduces the risk of IgE production induced by respiratory and oral allergens

Large size dna in vitro and in vivo delivery using chitosan transfection

In modern medicine, many diseases can be treated using gene therapy, which requires a DNA delivery system to prevent DNA from degradation and to transport it to the cells. Liposomal reagents are expensive for such a therapy and new inexpensive biodegradable DNA carriers are required. Chitosan (Ch) is a cationic polymer with promising potency for gene delivery. Some Ch derivatives have been shown to efficiently transfect mammalian cells in vitro. However, there are many inconsistencies in the literature concerning the effectiveness of Ch systems for in vivo gene transfer. The aim of this work was to develop a Ch-based vector for in vivo delivery of a large-size DNA fragment coding for the far-red fluorescent protein (RFP). Among several Ch derivatives, hexanoyl-Ch (HCh) with a molecular weight of 20 kDa effectively transfected cells in vitro. Intratumoural injection of polyplexes in colon and lung tumours resulted in local expression of RFP in tumours

Biodistribution of doxorubicin-loaded succinoyl chitosan nanoparticles in mice injected via intravenous or intranasal routes

Chitosan (Chi) is an extremely promising natural biopolymer with remarkable potency for the development of drug and vaccine delivery nanosystems. Various Chi derivatives are used to form nanoparticles (NPs) with unique properties. However, the efficacy of the therapy delivered by Chi NPs depends significantly on NP biodistribution in the body. The aim of this study was the analysis of biodistribution of NPs formed by succinoyl Chi and loaded with doxorubicin (SCNPDOX). We compared the distribution of free DOX and SCNP-DOX after intravenous (i.v.) and intranasal (i.n.) delivery into tumour-bearing mice. Distribution of DOX and SCNP-DOX was comparable after i.v. injection while they differed significantly after i.n. instillation

Express analysis of chitosan and its derivatives by gel electrophoresis

Chitosan is one of the most promising polymers for biomedical applications due to its unique properties, such as its biocompatibility, low toxicity, biodegradation, and the presence of reactive amino and hydroxyl groups. Analysis of physicochemical properties of chitosan and its derivatives is a time-consuming process and requires expensive equipment and large amounts of the sample. This paper proposes a method for express analysis of the molecular weight (MW), the degree of deacetylation (DD), the substitution degree (SD), and the charge of chitosan and its derivatives using agarose gel electrophoresis under acidic and neutral conditions with Coomassie staining. Positively charged chitosan samples required acidic Tris Acetate-EDTA (TAE) buffer to move in the gel. The electrophoretic mobility of chitosan depended on MW, DD, SD, and the chitosan charge. Based on the dependences obtained by the proposed method, the MW and DD of commercial chitosan samples were determined. Express analysis of chitosan and its derivatives in agarose gel can be used to monitor the reactions of chitosan modification and to analyse samples with unknown characteristics

Size-Dependent Biodistribution of Fluorescent Furano-Allocolchicinoid-Chitosan Formulations in Mice

The aim of this study was to compare the biodistribution in mice of functionalized rhodamine B (Rh) labeled colchicine derivative furano-allocolchicinoid (AC, 6) either conjugated to 40 kDa chitosan (AC-Chi, 8) or encapsulated into chitosan nanoparticles (AC-NPs). AC-NPs were formed by ionotropic gelation and were 400–450 nm in diameter as estimated in mice by dynamic light scattering and confocal microscopy. AC-Chi and AC-NPs preserved the specific colchicine activity in vitro. AC preparations were once IV injected into C75BL/6 mice; muscles, spleen, kidney, liver, lungs, blood cells and serum were collected at 30 min, 2, 5, 10, and 20 h post injection. To analyze the distribution of the furano-allocolchicinoid preparations in body liquids and tissues, Rh was measured directly in sera or extracted by acidic ethanol from tissue homogenates. Preliminary Rh extraction rate was estimated in vitro in tissue homogenates and was around 25–30% from total quantity added. After in vivo injection, AC-NPs were accumulated more in liver and spleen, while less in kidney and lungs in comparison with free AC and AC-Chi. Therefore, incorporation of colchicine derivatives as well as other hydrophobic substances into nano/micro sized carriers may help redistribute the drug to different organs and, possibly, improve antitumor accumulation

A Facile Synthetic Approach to Nonracemic Substituted Pyrrolo-allocolchicinoids Starting from Natural Colchicine

A six-step semisynthetic approach towards chiral nonracemic pyrrolo-allocolchicinoids starting from naturally occurring colchicine was developed. The synthetic scheme includes an electrocyclic tropolone ring contraction to afford allocolchicinic acid followed by the Curtius reaction, giving the corresponding aniline. The Sandmeyer reaction and copper-mediated hydrazination gave hydrazine-substituted allocolchicine. This was introduced into the Fischer indole synthesis, affording libraries of regioisomeric indole-based allocolchicine congeners