Trained immunity induction by the inactivated mucosal vaccine MV130 protects against experimental viral respiratory infections.

MV130 is an inactivated polybacterial mucosal vaccine that confers protection to patients against recurrent respiratory infections, including those of viral etiology. However, its mechanism of action remains poorly understood. Here, we find that intranasal prophylaxis with MV130 modulates the lung immune landscape and provides long-term heterologous protection against viral respiratory infections in mice. Intranasal administration of MV130 provides protection against systemic candidiasis in wild-type and Rag1-deficient mice lacking functional lymphocytes, indicative of innate immune-mediated protection. Moreover, pharmacological inhibition of trained immunity with metformin abrogates the protection conferred by MV130 against influenza A virus respiratory infection. MV130 induces reprogramming of both mouse bone marrow progenitor cells and in vitro human monocytes, promoting an enhanced cytokine production that relies on a metabolic shift. Our results unveil that the mucosal administration of a fully inactivated bacterial vaccine provides protection against viral infections by a mechanism associated with the induction of trained immunity.We are grateful to members of the D.S. laboratory for discussions and critical reading of the manuscript. We thank the CNIC facilities and personnel for assistance. P.B. is funded by grant BES-2014-069933 (‘‘Ayudas para Contratos Predoctorales para la Formacio´ n de Doctores 2014’’) from the Spanish Ministry of Economy, Industry and Competitiveness (MINECO). L.C. was a recipient of a European Respiratory Society Fellowship (RESPIRE2-2013- 3708). G.D. is supported by a European Molecular Biology Organization Long-term Fellowship (ALTF 379-2019). This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sk1odowska-Curie grant agreement No. 892965. Work in the D.S. laboratory is funded by the CNIC; by the European Research Council (ERC-2016-consolidator grant 725091); by the European Commission (635122-PROCROP H2020); by Ministerio de Ciencia e Innovacio´ n (MICINN), Agencia Estatal de Investigacio´ n (AEI), and Fondo Europeo de Desarrollo Regional (FEDER) (SAF2016-79040-R); by AEI (PID2019-108157RB); by Comunidad de Madrid (B2017/BMD-3733 Immunothercan-CM); by FIS-Instituto de Salud Carlos III, MICINN and FEDER (RD16/0015/0018-REEM); by a collaboration agreement with Inmunotek; by Atresmedia (Constantes y Vitales prize); by Fundacio´ La Marato´ de TV3 (201723); and by Fondo Solidario Juntos (Banco Santander). The CNIC is supported by the Instituto de Salud Carlos III, the MICINN, and the Pro CNIC Foundation.S

P2X7 receptor induces mitochondrial failure in monocytes and compromises NLRP3 inflammasome activation during sepsis

International audienceSepsis is characterized by a systemic inflammatory response followed by immunosuppres-sion of the host. Metabolic defects and mitochondrial failure are common in immunocom-promised patients with sepsis. The NLRP3 inflammasome is important for establishing an inflammatory response after activation by the purinergic P2X7 receptor. Here, we study a cohort of individuals with intra-abdominal origin sepsis and show that patient monocytes have impaired NLRP3 activation by the P2X7 receptor. Furthermore, most sepsis-related deaths are among patients whose NLRP3 activation is profoundly altered. In monocytes from sepsis patients, the P2X7 receptor is associated with mitochondrial dysfunction. Furthermore, activation of the P2X7 receptor results in mitochondrial damage, which in turn inhibits NLRP3 activation by HIF-1α. We show that mortality increases in a mouse model of sepsis when the P2X7 receptor is activated in vivo. These data reveal a molecular mechanism initiated by the P2X7 receptor that contributes to NLRP3 impairment during infection

Purinergic P2X7 receptor expression increases in leukocytes from intra-abdominal septic patients

Inflammation is a tightly coordinated response of the host immune system to bacterial and viral infections, triggered by the production of inflammatory cytokines. Sepsis is defined as a systemic inflammatory response followed by immunosuppression of the host and organ dysfunction. This imbalance of the immune response increases the risk of mortality of patients with sepsis, making it a major problem for critical care units worldwide. The P2X7 receptor plays a crucial role in activating the immune system by inducing the activation of peripheral blood mononuclear cells. In this study, we analyzed a cohort of abdominal origin septic patients and found that the expression of the P2X7 receptor in the plasma membrane is elevated in the different subsets of lymphocytes. We observed a direct relationship between the percentage of P2X7-expressing lymphocytes and the early inflammatory response in sepsis. Additionally, in patients whose lymphocytes presented a higher percentage of P2X7 surface expression, the total lymphocytes populations proportionally decreased. Furthermore, we found a correlation between elevated soluble P2X7 receptors in plasma and inflammasome-dependent cytokine IL-18. In summary, our work demonstrates that P2X7 expression is highly induced in lymphocytes during sepsis, and this correlates with IL-18, along with other inflammatory mediators such as IL-6, IL-8, and procalcitonin

The Bacterial Mucosal Immunotherapy MV130 Protects Against SARS-CoV-2 Infection and Improves COVID-19 Vaccines Immunogenicity

COVID-19-specific vaccines are efficient prophylactic weapons against SARS-CoV-2 virus. However, boosting innate responses may represent an innovative way to immediately fight future emerging viral infections or boost vaccines. MV130 is a mucosal immunotherapy, based on a mixture of whole heat-inactivated bacteria, that has shown clinical efficacy against recurrent viral respiratory infections. Herein, we show that the prophylactic intranasal administration of this immunotherapy confers heterologous protection against SARS-CoV-2 infection in susceptible K18-hACE2 mice. Furthermore, in C57BL/6 mice, prophylactic administration of MV130 improves the immunogenicity of two different COVID-19 vaccine formulations targeting the SARS-CoV-2 spike (S) protein, inoculated either intramuscularly or intranasally. Independently of the vaccine candidate and vaccination route used, intranasal prophylaxis with MV130 boosted S-specific responses, including CD8+-T cell activation and the production of S-specific mucosal IgA antibodies. Therefore, the bacterial mucosal immunotherapy MV130 protects against SARS-CoV-2 infection and improves COVID-19 vaccines immunogenicity.CF was supported by AECC Foundation (INVES192DELF) and is currently funded by the Miguel Servet program (ID: CP20/00106) (ISCIII). IH-M receives the support of a fellowship from la Caixa Foundation (ID 100010434, fellowship code: LCF/BQ/IN17/11620074) and from the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement no. 713673. AJ-C is a postgraduate fellow of the City Council of Madrid at the Residencia de Estudiantes (2020–2021). GD is supported by an European Molecular Biology Organization (EMBO) Long-term fellowship (ALTF 379-2019). This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No. Project number 892965. OL and JA-C acknowledge Comunidad de Madrid (Tec4Bio-CM, S2018/NMT-4443, FEDER). Work in OL laboratory was funded by CNIO with the support of the projects Y2018/BIO4747 and P2018/NMT4443 from Comunidad de Madrid and co-funded by the European Social Fund and the European Regional Development Fund. The CNIO is supported by the Instituto de Salud Carlos III (ISCIII). Work at CNB and CISA is funded by the Spanish Health Ministry, Instituto de Salud Carlos III (ISCIII), Fondo COVID-19 grant COV20/00151, and Fondo Supera COVID-19 (Crue Universidades-Banco Santander) (to JG-A). Work in the DS laboratory is funded by the CNIC; by the European Research Council (ERC-2016-Consolidator Grant 725091); by Agencia Estatal de Investigación (PID2019-108157RB); by Comunidad de Madrid (B2017/BMD-3733 Immunothercan-CM); by Fondo Solidario Juntos (Banco Santander); by a research agreement with Inmunotek S.L.; and by Fundació La Marató de TV3 (201723). The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the MICINN, and the Pro CNIC Foundation.Peer reviewe

Evaluación de estrategias diagnósticas de la agalaxia contagiosa en el ganado caprino / Joaquín Amores Iniesta; directores, Christian de la Fe Rodríguez, Antonio Sánchez López, Antonio Contreras de Vera.

Texto en español, francés e inglés.Tesis-Universidad de Murcia.Consulte la tesis en: BCA. GENERAL. ARCHIVO UNIVERSITARIO. TM 4293

Effective cancer immunotherapy by natural mouse conventional type-1 dendritic cells bearing dead tumor antigen

Background: The manipulation of dendritic cells (DCs) for cancer vaccination has not reached its full potential, despite the revolution in cancer immunotherapy. DCs are fundamental for CD8+ T cell activation, which relies on cross-presentation of exogenous antigen on MHC-I and can be fostered by immunogenic cancer cell death. Translational and clinical research has focused on in vitro-generated monocyte-derived DCs, while the vaccination efficacy of natural conventional type 1 DCs (cDC1s), which are associated with improved anti-tumor immunity and specialize on antigen cross-presentation, remains unknown. Methods: We isolated primary spleen mouse cDC1s and established a protocol for fast ex vivo activation and antigen-loading with lysates of tumor cells that underwent immunogenic cell death by UV irradiation. Natural tumor antigen-loaded cDC1s were transferred and their potential for induction of endogenous CD8+ and CD4+ T cell responses in vivo, cancer prevention and therapy were assessed in three grafted cancer models. Further, we tested the efficacy of natural cDC1 vaccination in combination and comparison with anti-PD-1 treatment in two "wildtype" tumor models not expressing exogenous antigens. Results: Herein, we reveal that primary mouse cDC1s ex vivo loaded with dead tumor cell-derived antigen are activated and induce strong CD8+ T cell responses from the endogenous repertoire upon adoptive transfer in vivo through tumor antigen cross-presentation. Notably, cDC1-based vaccines enhance tumor infiltration by cancer-reactive CD8+ and CD4+ T cells and halt progression of engrafted cancer models, including tumors that are refractory to anti-PD-1 treatment. Moreover, combined tumor antigen-loaded primary cDC1 and anti-PD-1 therapy had strong synergistic effects in a PD-1 checkpoint inhibition susceptible cancer model. Conclusions: This preclinical proof-of-principle study is first to support the therapeutic efficacy of cancer immunotherapy with syngeneic dead tumor cell antigen-loaded mouse cDC1s, the equivalents of the human dendritic cell subset that correlates with beneficial prognosis of cancer patients. Our data pave the way for translation of cDC1-based cancer treatments into the clinic when isolation of natural human cDC1s becomes feasible

Efficacy of sodium dodecyl sulfate for the inactivation of mycoplasmas in goat colostrum

En los rebaños de pequeños rumiantes crónicamente afectados de agalaxia contagiosa, los animales de reposición adquieren la infección a través de la ingestión de leche y calostro procedente de hembras infectadas que no muestran síntomas de la enfermedad. Como alternativa a los tradicionales métodos térmicos como la pasteurización, este estudio examina el efecto de diferentes concentraciones de dodecil sulfato de sodio (DSS) sobre la viabilidad de Mycoplasma agalactiae y Mycoplasma mycoides subsp. capri en calostro caprino. Pos- teriormente al tratamiento y antes de incubar las muestras de calostro contaminado, el DSS fue extraído o no de las muestras para evaluar el efecto de la prolongación del tratamiento durante el periodo de incubación. El tratamiento durante diez minutos con DSS al 0,1% fue inefectivo contra ambas especies de micoplasmas. Tratando el calostro con DSS al 1% no se observó crecimiento cuando el detergente estuvo presente durante el periodo de incubación, mientras que se mantuvo la viabilidad de los micoplasmas cuando el DSS fue extraído justo antes de este periodo. Nuestros hallazgos indican que la duración del tratamiento del calostro con DSS es crítico para su efecto de inactivación sobre estos micoplasmas.ABSTRACT: In small ruminant herds chronically affected with contagious agalactia, replacement animals acquire the infection by suckling the milk and colostrum of infected dams that show no symptoms of the disease. As an alterative to traditional heat treatments such as pasteurisation, this study examines the effect of different concentrations of sodium dodecyl sulfate (SDS) on the viability of Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri in goat colostrum. After the treatment and before incubating contaminated colostrum samples, SDS was removed or not from the samples to assess the effect of prolonged treatment during the incubation period. Ten minutes of treatment with 0.1% SDS were ineffective against both species of mycoplasma. When SDS was used at 1%, no growth was observed when the detergent was present during the incubation period yet viable colonies appeared when SDS was removed just before this period. Our findings indicate that the duration of colostrum SDS treatment is critical for its inactivation effect on these mycoplasmas

Sanitary involvements of management of colostrum in caprine herds

El calostro supone la primera fuente de inmunidad para los rumiantes y por tanto determina su resistencia a enfermedades durante las primeras horas de vida. No obstante, la ingesta de calostro puede suponer en sí misma una vía de transmisión de diversas enfermedades, como la paratuberculosis, la artritis-encefalitis caprina o la agalaxia contagiosa. Este riesgo puede evitarse siguiendo un régimen de lactancia artificial con unas adecuadas pautas de manejo del calostro. Entre dichas pautas, el tratamiento del calostro supone un punto crítico. En este sentido, se han empleado los tratamientos térmicos para higienizar el calostro, observándose resultados diversos en la viabilidad de distintos microorganismos. Al mismo tiempo, se debe considerar el efecto negativo del calor sobre la composición nutricional del calostro, principalmente la pérdida de inmunoglobulinas. Como alternativa a los tratamientos térmicos, a nivel experimental, se han empleado métodos como la adición de dodecil sulfato de sodio, capaz de inactivar el virus del síndrome de inmunodeficiencia humana en leche, y otros procesos como la liofilización o el uso de altas presiones. Previamente a la aplicación práctica de las diferentes opciones de tratamiento del calostro se deberá considerar su viabilidad económica y su factibilidad en la explotación.ABSTRACT: Colostrum represents the first source of immunity for the ruminants, and thus determines its resistance to disease during the first hours of life. However, colostrum intake could be itself the way of transmition of several diseases, as paratuberculosis, caprine arthritis-encephalytis, or micoplasmosis like contagious agalactia. This risk could be avoided by means of an artificial rearing program which should include correct management practices for colostrum. Between them, the treatment of colostrum represents a critical point. In this sense, thermic treatments have been used to higienitize colostrum, showing different results about microorganism viability. Nevertheless, it should be considered the negative effect of these treatments over nutritional components of colostrum, particularly the loss of immunoglobulines. As an alternative to thermic treatments, there have been experimentally assayed other methods as the addition of sodium dodecyl sulphate, which inactivates AIDS virus in breast milk, and others as liophilization or high pression methods. In this works, apart from the effect of the treatment should be also taken into account its economical viability and on-farm feasibility

Etiological and epidemiological particularities of ovine contagious agalactia: similarities and differences with goats

Este trabajo describe las diferencias existentes en la presentación de la agalaxia contagiosa, síndrome infectocontagioso causado por varias especies del genero Mycoplasma spp., en el ganado ovino y caprino. Las particularidades etiológicas y epidemiológicas de la infección crónica en la cabra, y la ausencia de datos similares en el ovino, evidencian la necesidad de realizar nuevos trabajos que determinen si las diferencias observadas en referencia a la presencia de portadores auriculares en los rebaños o la participación de los sementales en la difusión de la enfermedad son fruto de la escasez de trabajos científicos o por el contrario, establecen verdaderas diferencias sobre las que desarrollar estrategias dirigidas de control en función de la especie de rumiante afectada.ABSTRACT Contagious agalactia (CA) is an infectious syndrome caused by several species of Mycoplasma spp. which affects small ruminants. The aim of this review is to describe the main differences noted between the disease in sheep and goats, especially with regard to its etiologic and epidemiological peculiarities. Thus, the presence of asymptomatic auricular carriers of CA-causing mycoplasmas and the stud’s role in the transmission of the disease are well known in goats, but scarcely evaluated in sheep. Further studies are needed to determine if these differences are real or if they are due to the shortage of scientific work in this matter