Demostraciones prácticas para la promoción de las titulaciones de Grado en Química y Grado en Ingeniería Química

Memoria ID12-0251. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2012-2013

Colaboración universidad/empresa para la promoción de las titulaciones de grado en química y grado en ingeniería química

Memoria ID-0018. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2013-2014

Modulation by Anisakis simplex antigen of inflammatory response generated in experimental autoimmune encephalomyelitis

The impact of immunization with Anisakis simplex larval antigen on the occurrence and progression of experimental autoimmune encephalomyelitis (EAE) induced in mice was studied. C57BL/6J mice were immunized with the MOG35-55 peptide and one batch was treated with A. simplex total larval antigen on days 1, 8, 10 and 12 after EAE induction. Significantly higher values were obtained in the EAE clinical parameters of the antigen-treated group. Likewise, there was a significant decrease in the weights of the animals. Anisakis-treatment produced a significant decrease in anti-MOG35-55 specific IgG1 on day 21. On day 14 there was an increase in serum IL-2, IL-6, IL-10, IL-17A, and TGF-β in the treated group. On day 21, a decrease in IL-4, IL-6, TNF-α, TGF-β was observed. All brain determinations were made on day 21. The treatment decreased values of IL-6, IL-10, IL-17A and TNF-α. A. simplex antigen caused a significantly higher incidence of EAE and an advance in the appearance of the disease manifestations. However, treatment with the antigen was able to cause a decrease in proinflammatory cytokines (IL-6, IL-17A, and TNF-α) in nervous tissue that could establish a future preventive scenario for myelin damage.SANTANDER/COMPLUTENSEDepto. de Microbiología y ParasitologíaFac. de FarmaciaTRUEpu

Rodent Models for the Study of Soil-Transmitted Helminths: A Proteomics Approach

Soil-transmitted helminths (STH) affect hundreds of millions worldwide and are some of the most important neglected tropical diseases in terms of morbidity. Due to the difficulty in studying STH human infections, rodent models have become increasingly used, mainly because of their similarities in life cycle. Ascaris suum and Trichuris muris have been proven appropriate and low maintenance models for the study of ascariasis and trichuriasis. In the case of hookworms, despite most of the murine models do not fully reproduce the life cycle of Necator americanus, their proteomic similarity makes them highly suitable for the development of novel vaccine candidates and for the study of hookworm biological features. Furthermore, these models have been helpful in elucidating some basic aspects of our immune system, and are currently being used by numerous researchers to develop novel molecules with immunomodulatory proteins. Herein we review the similarities in the proteomic composition between Nippostrongylus brasiliensis, Heligmosomoides polygyrus bakeri and Trichuris muris and their respective human counterpart with a focus on the vaccine candidates and immunomodulatory proteins being currently studied.Miguel Servet FellowInstituto de Salud Carlos IIIDepto. de Microbiología y ParasitologíaFac. de FarmaciaTRUEpu

Allergenicity of vertebrate tropomyosins: Challenging an immunological dogma

With the exception of tilapia tropomyosin, other anecdotic reports of tropomyosin recognition of vertebrate origin are generally not accompanied by clinical significance and a dogmatic idea is generally accepted about the inexistence of allergenicity of vertebrate tropomyosins, based mainly on sequence similarity evaluations with human tropomyosins. Recently, a specific work-up of a tropomyosin sensitised patient with seafood allergy, demonstrated that the IgE-recognition of tropomyosin from different fish species can be clinically relevant. We hypothesise that some vertebrate tropomyosins could be relevant allergens. The hypothesis is based on the molecular evolution of the proteins and it was tested by in silico methods. Fish, which are primitive vertebrates, could have tropomyosins similar to those of invertebrates. If the hypothesis is confirmed, tropomyosin should be included in different allergy diagnosis tools to improve the medical protocols and management of patients with digestive or cutaneous symptoms after fish intake.Universidad Complutense de MadridDepto. de Microbiología y ParasitologíaFac. de FarmaciaTRUEpu

Are fish tropomyosins allergens?

Universidad Complutense de MadridDepto. de Microbiología y ParasitologíaFac. de FarmaciaTRUEpu

Possible Allergenic Role of Tropomyosin in Patients with Adverse Reactions after Fish Intake

In a recent case report, patient's anti-fish tropomyosin IgE was associated with gastrointestinal symptoms. We aimed to demonstrate on a wider scale that the panallergen tropomyosin should not be limited to invertebrate species and that clinically relevant reactions could be elicited by vertebrate tropomyosin. On the whole, 19 patients with adverse reactions after fish intake and showing negative skin tests with commercial fish extracts were included. Fish tropomyosin was recognized by 10/19 patients' IgE by immunoblotting. All patients with gastrointestinal complaints after fish intake (6/6) showed an IgE band matching with tropomyosin. Cod, albacore, and swordfish tropomyosins were recognized by most patients although 3/10 patients did not claim adverse reactions to these fish species. Immunoblotting with a battery of antigens from different fish species have a high yield of positivity at a band matching with tropomyosin molecular weight, even if they have not been claimed to be causative agents of symptoms. Tropomyosin is therefore a good candidate to be investigated as a clinically relevant fish allergen in patients who report adverse reactions after fish intake.Depto. de Microbiología y ParasitologíaFac. de FarmaciaTRUEpu

New insights into the allergenicity of tropomyosin: a bioinformatics approach

The invertebrate panallergen tropomyosin is a protein with an extremely simple folding. This makes it a perfect target for investigating structural differences between invertebrate and vertebrate tropomyosins, which are not considered allergenic. Phylogenetic and sequence analyses were conducted in order to explore the differences in primary structure between several tropomyosins and to promote an experimental development in the field of food allergy, based on the study of tropomyosin. The phylogenetic analyses showed that tropomyosin is a useful evolutionary marker. The phylogenetic trees obtained with tropomyosin were not always phylogenetically correct, but they might be useful for allergen avoidance by tropomyosin allergic individuals. Sequence analyses revealed that the probability of alpha helix folding in invertebrate tropomyosins was lower than in all the studied vertebrate ones, except for the Atlantic bluefin tuna Thunnus thynnus tropomyosin. This suggested that the lack of alpha helix folding may be involved in the immunogenicity of tropomyosins. More specifically, the regions adjacent to the positions 133-135 and 201 of the invertebrate tropomyosins, presented lower probability of alpha helix folding than those of vertebrates and are candidates to be responsible for their allergenicity.Fundación Ramón ArecesDepto. de Microbiología y ParasitologíaFac. de FarmaciaTRUEpu

Anisakis simplex: Immunomodulatory effects of larval antigens on the activation of Toll like Receptors

Aims: The objective of this investigation is to evaluate the mechanisms Anisakis simplex employs to modify its host immune system, regarding the larval antigens interactions with Toll-Like-Receptors (TLRs). Methods and Results: In a previous study, we described that the stimulation of bone marrow derived dendritic cells (BMDCs) with A. simplex larval antigens drive an acute inflammatory response in BALB/c mice, but a more discrete and longer response in C57BL/6J. Moreover, when A. simplex larval antigens were combined with TLR agonists (TLR 1/2–9), they modified mainly TLR2, TLR4 and TLR9 agonists responses in both mice strains, and also TLR3, TLR5 and TLR7 in BALB/c. Antigen-presenting ability was analyzed by the detection of CD11c + cells expressing surface markers (CD80-86, MHC I-II), intracellular cytokines (IL-10, IL-12, TNF-α) and intracellular proteins (Myd88, NF-κβ) by Flow Cytometry. Secreted IL-10 was measured by ELISA. Conclusion: Our findings confirm not only that the host genetic basis plays a role in the development of a Th2/Th1/Treg response, but also it states A. simplex larval antigens present specific mechanisms to modify the innate response of the host. As allergies share common pathways with the immune response against this particular helminth, our results provide a better understanding into the specific mechanisms of A. simplex allergy related diseases.Fundación Ramón Areces, SpainDepto. de Microbiología y ParasitologíaFac. de FarmaciaTRUEpu