SUMMARY OF RESEARCH

Identification of antigens and immunostimultatory molecules candidates for inmunotherapy against trypanosomatid pathogens

Parasites of the Trypanosomatidae family are the etiological agent of leishmaniosis and American tripanosomiasis (Chagas? Disease) characterized by presenting unspecific clinical symptoms together with an alteration of the host immune system. The parasites are transmitted by insect vectors, producing high morbidity and mortality rates. It has been estimated that 300 million persons live in endemic areas and that there are 500.000 new cases each year. Nowadays iin development countries there has been a high increase in the rate of infection due probably to the existence of additional transmission ways, such as blood transfusions (T. cruzi). As it occurs for Leishmania infantum, there is a higher rate of canine infections and an association with immunosupression states.

Given that existing chemotherapy is toxic and not very specific, vaccines, despite the difficulties involved in their developing, are probably the most effective way of responding to the world-wide important problem presented by these infectious pathologies. Although the protective antiparasitic response mechanisms of the host are not fully understood, it is clear, for both diseases, that effective protection requires induction of a multiple immune response incorporating serialized cellular activation and antibody production with a balance towards Th1 and induction of parasite´s antigen-specific CTLs. Undoubtedly, it requires immunization with suitable antigenic protein/s associated to carrier molecules, stimulators/modulators of the immune response. The genetic vaccines or DNA vaccines, as well as recombinant proteins, may be an excellent way of inducing this type of multiple response and of generating effective protection.

The scientific objective of our laboratory is focused on identifying and studying at molecular and immunological level specific proteins from Leishmania and Trypanosoma cruzi which are antigenic molecules and candidates for vaccines, as well as their encoding genes. Likewise, we have special interest in the characterization of biological molecules with immunostimulatory capability. The final aim of the laboratory is directed to analyze the capacity of the chimeric recombinant proteins or DNA vectors to limit the pathological consequences of leishmaniosis and trypanosomiasis and to induce protection against parasite´s infection

Study at molecular level of the transposition and integration mechanism of L1Tc mobile element from T. cruzi. Analysis of its functionality

The genome of a large number of prokaryotic and eukaryotic organisms contains a surprisingly high proportion of repeated long and short interspersed nucleotide elements (LINE and SINE) DNA sequences with the potential capacity to transpose in the host genome. These mobile elements have been considered for decades as selfish elements. However, at present these sequences have been taken as important elements to generate variations in genome structure and expression associated to species evolution. Although the biological properties of these sequences (regulation, replication and interaction with their host organisms) are not well understood, the available data indicate that they represent challenging and fascinating biological elements playing crucial functional roles. In this context the resolution of these functions and mechanisms will be essential to know how the repeated elements dialogue to achieve the concrete modeling of the genetic information and consequently to fully understand the biology of the host organisms.

The genome of the parasite pathogen Trypanosoma cruzi has a large proportion of repeated and dispersed DNA sequences which share common characteristics with SINE sequences from higher eukaryotes, and a LINE type element known as L1Tc encoding for the enzymes required for its retrotransposition. These retrotransposon sequences have been linked to the high plasticity observed in the T. cruzi genome and furthermore, seem to be involved in the chromosomal organization and control of the expression of specific genes, crucial for the pathogenicity and survival of the parasite.

The objectives of our laboratory are focused on the characterization of the molecular and functional characteristics of the different polypeptides encoded by the LINE-L1Tc element from T. cruzi as well as its putative regulatory sequences. Likewise, we are interested in determining at molecular level, the retrotransposition and integration mechanisms of this mobile element and in the identification of the effect that de novo retrotransposition events have on the genome of the aforementioned parasite pathogen.

FUNDING AGENCIES LAST 5 YEARS

- Desarrollo de nuevas herramientas de prevención y diagnóstico de infecciones resistentes a antibióticos por Klebsiella pneumoniae. PROYECTO, PN2022 -Proy en colaboración público-privada - MRR, Ref: CPP2022-009987, (2023 - 2026).

- Control de la leishmaniosis: ¿Perfil funcional, fenotípico y de expresión génica de la respuesta inmunológica celular antígeno-específica asociada al control de la infección por Leishmania infantum?. PROYECTO, J.A.- Retos de la sociedad andaluza 2021, Ref: P21_00852, (2022 - 2025).

- DESARROLLO Y FABRICACIÓN SOLUCIONES Y PREPARACIÓN ANTE LA CRISIS SANITARIA MUNDIAL POR LA RESISTENCIA A ANTIBIÓTICOS - AMReady. PROYECTO, PN2021 - Proy de I+D+i líneas estratégicas (PLEC), Ref: PLEC2021-008078, (2021 - 2024).

- VALORIZACIÓN DE NUEVAS VACUNAS CONTRA PATÓGENOS RESPIRATORIOS PARA LA REDUCCIÓN DEL USO DE ANTIBIÓTICOS EN PORCINO. PROYECTO, PN2017 -RETOS COLABORACION - PRG. RETOS DE LA SOC., Ref: RTC-2017-6601-2, (2018 - 2020).

- Red de investigación colaborativa en enfermedades tropicales RICET. PROYECTO, PN2016 - REDES TEMATICAS INV. COOPERATIVA, Ref: RD16/0027/0005, (2017 - 2021).

- BIOMARCADORES INMUNOLOGICOS ANTIGENO-ESPECIFICOS (CELULAS T) ASOCIADOS AL CONTROL DE LA INFECCION POR T. CRUZI.. PROYECTO, PN2016 - PROY I+D+I - PRG. RETOS DE LA SOCIEDAD, Ref: SAF2016-81003-R, (2016 - 2020).

PUBLICATIONS LAST 5 YEARS

-Garcia-Castro A; Egui Machado A; Thomas Carazo MC; Lopez Lopez Manuel Carlos, Humoral and Cellular Immune Response in Asymptomatic Dogs with Visceral Leishmaniasis: A Review., Vaccines, 2022, Vol. 10: 947, ARTÍCULO DE REVISIÓN, Id:894194

-Alonso-Vega, C.; Urbina, J.A.; Sanz, S.; Pinazo, M.J.; Pinto, J.J.; Gonzalez, V.R.; Rojas, G.; Ortiz, L.; Garcia, W.; Lozano, D.; Soy, D.; Maldonado, R.A.; Nagarkatti, R.; Debrabant, A.; Schijman, A.; Thomas, M.C.; López, M.C.; Michael, K.; Ribeiro, I.; Gascon, J.; Torrico, F.; Almeida, I.C., New chemotherapy regimens and biomarkers for Chagas disease: The rationale and design of the TESEO study, an open-label, randomised, prospective, phase-2 clinical trial in the Plurinational State of Bolivia, BMJ Open, 2021, Vol. 11: 12-e052897, ARTÍCULO, Id:868154

-Mateus J; Nocua P; Lasso P; López MC; Thomas MC; Egui A; Cuervo C; González JM; Puerta CJ; Cuellar A., CD8+ T cell response quality is related to parasite control in an animal model of single and mixed chronic Trypanosoma cruzi infections, Frontiers in cellular and infection microbiology, 2021, Vol. 11: 723121, ARTÍCULO, Id:865569

-Gómez, I.; Thomas, M.C.; Palacios, G.; Egui, A.; Carrilero, B.; Simón, M.; Valladares, B.; Segovia, M.; Carmelo, E.; López, M.C., Differential Expression of Immune Response Genes in Asymptomatic Chronic Chagas Disease Patients Versus Healthy Subjects, Frontiers in cellular and infection microbiology, 2021, Vol. 11: 722984, ARTÍCULO, Id:854449

-Gómez, I.; López, M.C.; Rastrojo, A.; Lorenzo-Díaz, F.; Requena, J.M.; Aguado, B.; Valladares, B.; Thomas, M.C., Variability of the Pr77 sequence of L1Tc retrotransposon among six T. cruzi strains belonging to different discrete typing units (DTUs), Acta Tropica, 2021, Vol. 222: 106053, ARTÍCULO, Id:852664

-Alonso-Padilla, J.; López, M.C.; Esteva, M.; Zrein, M.; Casellas, A.; Gómez, I.; Granjon, E.; Méndez, S.; Benítez, C.; Ruiz, A.M.; Sanz, S.; Gascón, J.; Thomas, M.C.; Pinazo, M.J.; Abril, M.; de Noya, B.A.; Jorge, T.A.; Chatelain, E.; Grijalva, M.J.; Guhl, F.; Hasslocher-Moreno, A.M.; Luquetti, A.O.; Noya, O.; Ramsey, J.M.; Ribeiro, I.; Longhi, S.A.; Schijman, A.G.; Sosa-Estani, S.; Torrico, F.; Viotti, R., Serological reactivity against T. cruzi-derived antigens: Evaluation of their suitability for the assessment of response to treatment in chronic Chagas disease., Acta Tropica, 2021, Vol. 221: 105990, ARTÍCULO, Id:851634

-Pérez-Antón E; Egui A; Thomas MC; Carrilero B; Simón M; López-Ruz MÁ; Segovia M; López MC., A proportion of CD4+ T cells from patients with chronic Chagas disease undergo a dysfunctional process, which is partially reversed by benznidazole treatment., PLoS Neglected Tropical Diseases, 2021, Vol. 15: e00090592-e0009059, ARTÍCULO, Id:841082

-Cantos-Barreda, A.; Escribano, D.; Siriyasatien, P.; Cerón, J.J.; Thomas, M.C.; Afonso-Lehmann, R.N.; López, M.C.; Bernal, L.J.; Phumee, A.; Lubas, G.; Martínez-Subiela, S., Detection of Leishmania infantum DNA by real-time PCR in saliva of dogs, Comparative Immunology, Microbiology and Infectious Diseases, 2020, Vol. 73: 101542, ARTÍCULO, Id:815998

-Gómez, I.; Rastrojo, A.; Lorenzo-Diáz, F.; Sánchez-Luque, F.J.; MacIás, F.; Aguado, B.; Valladares, B.; Requena, J.M.; López, M.C.; Thomas, M.C., Trypanosoma cruzi Ikiakarora (TcIII) Draft Genome Sequence, Microbiology Resource Announcements, 2020, Vol. 9: 27-e00453-20, ARTÍCULO, Id:815969

-Gómez, I.; Rastrojo, A.; Sanchez-Luque, F.J.; Lorenzo-Díaz, F.; Macías, F.; Valladares, B.; Aguado, B.; Requena, J.M.; López, M.C.; Carmen Thomas, M., Draft Genome sequence of the trypanosoma cruzi B. M. López strain (TCIA), isolated from a Colombian patient, Microbiology Resource Announcements, 2020, Vol. 9: 18-e00031-20, ARTÍCULO, Id:809266

-Alonso-Padillaid, J.; Abril, M.; de Noya, B.A.; Almeida, I.C.; Angheben, A.; Jorge, T.A.; Chatelain, E.; Esteva, M.; Gascón, J.; Grijalva, M.J.; Guhl, F.; Hasslochermoreno, A.M.; López, M.C.; Luquetti, A.; Noya, O.; Pinazo, M.J.; Ramsey, J.M.; Ribeiro, I.; Ruiz, A.M.; Schijman, A.G.; Sosa-Estani, S.; Carmen Thomas, M.; Torrico, F.; Zreinid, M.; Picado, A., Target product profile for a test for the early assessment of treatment efficacy in chagas disease patients: An expert consensus, PLoS Neglected Tropical Diseases, 2020, Vol. 14: 1-10, ARTÍCULO, Id:808851

-Cantos-Barreda, A.; Escribano, D.; Egui, A.; López, M.C.; Cerón, J.J.; Bernal, L.J.; Martínez-Subiela, S., Evaluation of the circadian rhythm of anti-Leishmania IgG2 and IgA antibodies in serum and saliva of dogs with clinical leishmaniosis, Comparative Immunology, Microbiology and Infectious Diseases, 2020, Vol. 68: 101389, ARTÍCULO, Id:804240

-Pérez-Antón, E.; Egui, A.; Thomas, M.C.; Simón, M.; Segovia, M.; López, M.C., Immunological exhaustion and functional profile of CD8+ T lymphocytes as cellular biomarkers of therapeutic efficacy in chronic Chagas disease patients, Acta Tropica, 2020, Vol. 202: 105242, ARTÍCULO, Id:804208

-Egui, A.; López, M.C.; Gómez, I.; Simón, M.; Segovia, M.; Thomas, M.C., Differential phenotypic and functional profile of epitope-specific cytotoxic CD8+ T cells in benznidazole-treated chronic asymptomatic Chagas disease patients, Biochimica et Biophysica Acta - Molecular Basis of Disease, 2020, Vol. 1866: 3-165629, ARTÍCULO, Id:790664

-Franco-Martínez, L.; Villar, M.; Tvarijonaviciute, A.; Escribano, D.; Bernal, L.J.; Cerón, J.J.; Thomas, M.d.C.; Mateos-Hernández, L.; Tecles, F.; de la Fuente, J.; López, M.C.; Martínez-Subiela, S., Serum proteome of dogs at subclinical and clinical onset of canine leishmaniosis, Transboundary and Emerging Diseases, 2020, Vol. 97: 318-327, ARTÍCULO, Id:774803

-Cantos-Barreda, A.; Escribano, D.; Egui, A.; Thomas, M.C.; López, M.C.; Tecles, F.; Bernal, L.J.; Cerón, J.J.; Martínez-Subiela, S., One-year follow-up of anti-Leishmania antibody concentrations in serum and saliva from experimentally infected dogs, International Journal for Parasitology, 2019, Vol. 49: 893-900, ARTÍCULO, Id:771543

-Egui, A.; Carmen Thomas, M.; Fernández-Villegas, A.; Pérez-Antón, E.; Gómez, I.; Carrilero, B.; del Pozo, Á.; Ceballos, M.; Andrés-León, E.; López-Ruz, M.Á.; Gainza, E.; Oquiñena, E.; Segovia, M.; López, M.C., A Parasite Biomarker Set for Evaluating Benznidazole Treatment Efficacy in Patients with Chronic Asymptomatic Trypanosoma cruzi Infection, Antimicrobial Agents and Chemotherapy, 2019, Vol. 63: 10-e02436-18, ARTÍCULO, Id:771537

-Simón, M.; Gil-Gallardo, L.J.; Asunción Iborra, M.; Carrilero, B.; López, M.C.; Romay-Barja, M.; Murcia, L.; Carmen Thomas, M.; Benito, A.; Segovia, M., An observational longitudinal study to evaluate tools and strategies available for the diagnosis of Congenital Chagas Disease in a non-endemic country, Acta Tropica, 2019, Vol. 199: 105127, ARTÍCULO, Id:770497

-Franco-Martínez, L.; Tvarijonaviciute, A.; Horvati¿, A.; Guillemin, N.; Bernal, L.J.; Bari¿ Rafaj, R.; Cerón, J.J.; Thomas, M.d.C.; López, M.C.; Tecles, F.; Martínez-Subiela, S.; Mrljak, V., Changes in saliva of dogs with canine leishmaniosis: A proteomic approach, Veterinary Parasitology, 2019, Vol. 272: 44-52, ARTÍCULO, Id:767572

-Egui A; Lasso P; Perez-Anton E; Thomas MC; Lopez MC, Dynamics of T cells repertoire during Trypanosoma cruzi infection and its post-treatment modulation., Current Medicinal Chemistry, 2019, Vol. 26: 1-22, ARTÍCULO, Id:756060

-Risueño, J.; Spitzová, T.; Bernal, L.J.; Muñoz, C.; López, M.C.; Thomas, M.C.; Infante, J.J.; Volf, P.; Berriatua, E., Longitudinal monitoring of anti-saliva antibodies as markers of repellent efficacy against Phlebotomus perniciosus and Phlebotomus papatasi in dogs, Medical and Veterinary Entomology, 2019, Vol. 33: 99-109, ARTÍCULO, Id:743780

-Egui A; Ledesma D; Pérez-Antón E; Montoya A; Gómez I; Robledo SM; Infante JJ; Vélez ID; López MC; Thomas MC, Phenotypic and Functional Profiles of Antigen-Specific CD4+ and CD8+ T Cells Associated With Infection Control in Patients With Cutaneous Leishmaniasis., Frontiers in cellular and infection microbiology, 2018, Vol. 8: 393-393, ARTÍCULO, Id:756009

-Ontoria, E.; Hernández-Santana, Y.E.; González-García, A.C.; López, M.C.; Valladares, B.; Carmelo, E., Transcriptional profiling of immune-related genes in Leishmania infantum-infected mice: Identification of potential biomarkers of infection and progression of disease, Frontiers in cellular and infection microbiology, 2018, Vol. 8: 197, ARTÍCULO, Id:727592

-Pérez-Antón, E.; Egui, A.; Thomas, M.C.; Puerta, C.J.; González, J.M.; Cuéllar, A.; Segovia, M.; López, M.C., Impact of benznidazole treatment on the functional response of Trypanosoma cruzi antigen-specific CD4+CD8+T cells in chronic Chagas disease patients, PLoS Neglected Tropical Diseases, 2018, Vol. 12: 5-e0006480, ARTÍCULO, Id:723199

-Macías, F.; Afonso-Lehmann, R.; López, M.C.; Gómez, I.; Carmen Thomas, M., Biology of trypanosoma cruzi retrotransposons: From an enzymatic to a structural point of view, Current Genomics, 2018, Vol. 19: 110-118, ARTÍCULO DE REVISIÓN, Id:715936

DOCTORAL THESES LAST 5 YEARS