Neutrophils which migrate to lymph nodes modulate CD4+ T cell response by a PD-L1 dependent mechanism

It is well known that neutrophils are rapidly recruited to a site of injury or infection and perform a critical role in pathogen clearance and inflammation. However, they are also able to interact with and regulate innate and adaptive immune cells and some stimuli induce the migration of neutrophils to lymph nodes (LNs). Previously, we demonstrated that the immune complex (IC) generated by injecting OVA into the footpad of OVA/CFA immunized mice induced the migration of OVA+ neutrophils to draining LNs (dLNs). Here we investigate the effects of these neutrophils which reach dLNs on CD4+ T cell response. Our findings here strongly support a dual role for neutrophils in dLNs regarding CD4+ T cell response modulation. On the one hand, the CD4+ T cell population expands after the influx of OVA+ neutrophils to dLNs. These CD4+ T cells enlarge their proliferative response, activation markers and IL-17 and IFN-γ cytokine production. On the other hand, these neutrophils also restrict CD4+ T cell expansion. The neutrophils in the dLNs upregulate PD-L1 molecules and are capable of suppressing CD4+ T cell proliferation. These results indicate that neutrophils migration to dLNs have an important role in the homeostasis of adaptive immunity. This report describes for the first time that the influx of neutrophils to dLNs dependent on IC presence improves CD4+ T cell response, at the same time controlling CD4+ T cell proliferation through a PD-L1 dependent mechanism.Fil: Castell, Sofía Daiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Harman, María Florencia. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Morón, Sergio Gabriel. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; ArgentinaFil: Maletto, Belkys Angélica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Pistoresi Palencia, María C.. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentin

Neutrophils Which Migrate to Lymph Nodes Modulate CD4+ T Cell Response by a PD-L1 Dependent Mechanism

It is well known that neutrophils are rapidly recruited to a site of injury or infection and perform a critical role in pathogen clearance and inflammation. However, they are also able to interact with and regulate innate and adaptive immune cells and some stimuli induce the migration of neutrophils to lymph nodes (LNs). Previously, we demonstrated that the immune complex (IC) generated by injecting OVA into the footpad of OVA/CFA immunized mice induced the migration of OVA+ neutrophils to draining LNs (dLNs). Here we investigate the effects of these neutrophils which reach dLNs on CD4+ T cell response. Our findings here strongly support a dual role for neutrophils in dLNs regarding CD4+ T cell response modulation. On the one hand, the CD4+ T cell population expands after the influx of OVA+ neutrophils to dLNs. These CD4+ T cells enlarge their proliferative response, activation markers and IL-17 and IFN-γ cytokine production. On the other hand, these neutrophils also restrict CD4+ T cell expansion. The neutrophils in the dLNs upregulate PD-L1 molecules and are capable of suppressing CD4+ T cell proliferation. These results indicate that neutrophils migration to dLNs have an important role in the homeostasis of adaptive immunity. This report describes for the first time that the influx of neutrophils to dLNs dependent on IC presence improves CD4+ T cell response, at the same time controlling CD4+ T cell proliferation through a PD-L1 dependent mechanism

Dietary fatty acids and inflammatory salivary markers in lichenoid diseases and cancer

Fil: Costantino, Evangelina. Universidad Nacional de Córdoba. Facultad de Odontología. Cátedra de Anatomía B; Argentina.Fil: Costantino, Evangelina. Universidad Nacional de Córdoba. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ciencias de la Salud; Argentina.Fil: Castell, Sofía Daiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina.Fil: Panico, René Luis. Universidad Nacional de Córdoba. Facultad de Odontología. Cátedra de Estomatología A; Argentina.Fil: Pascualini, María Eugenia. Universidad Nacional de Córdoba. Facultad de Ciencias Médicas. Cátedra de Biología Celular, Histología y Embriología; Argentina.Fil: Pascualini, María Eugenia, Pablo Gastón. Universidad Nacional de Córdoba. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ciencias de la Salud; Argentina.Fil: Pistoresi Palencia, María C. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina.Fil: Actis, Adriana Beatríz. Universidad Nacional de Córdoba. Facultad de Odontología. Secretaría de Ciencia y Técnica; Argentina.Fil: Actis, Adriana Beatriz. Universidad Nacional de Córdoba. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ciencias de la Salud; Argentina.Ácidos grasos dietarios y marcadores inflamatorios en saliva de pacientes con enfermedades liquenoides bucales y cáncer bucalCostantino E, Castell SD, Panico RL, Pasqualini ME, Pistoresi MC, Actis ABObjetivo: analizar la relación entre ácidos grasos (AG) dietarios y la concentración de marcadores inflamatorios en saliva de personas con enfermedades liquenoides bucales (ELB) y carcinoma bucal de células escamosas (CBCE).Pacientes y métodos: se incluyeron 31 personas con ELB (n=20) y CBCE (n=11), y 44 voluntarios sanos (S) (40-80 años en todos los grupos). Se completó una encuesta de frecuencia de consumo alimentario cualitativo-cuantitativo validada. Los datos se procesaron mediante Interfood v.1.3. De cada participante se obtuvo saliva mixta no estimulada para analizar citoquinas -interleuquinas 1&#946; (IL-1&#946;), 6 (IL-6) y factor de necrosis tumoral (TNF) (ELISA)- y eicosanoides -ácido 12-hidroxiheptadecatrienoico (12-HHT), ácido 13-hidroxioctadecadienoico (13-HODE), ácido 12 y 5 hidroxieicosatetraenoico (12 y 5-HETE)- (HPLC). Se empleó la prueba de Kruskal Wallis y el coeficiente de correlación de Spearman (p<0.05).Resultados: Los niveles de IL-6 y TNF fueron más altos en pacientes con lesión bucal que en S (p<0,001 y p=0,005, respectivamente). 13-HODE, 5 y 12-HETE fueron más altos en ELB que en CBCE, y sus niveles también fueron más altos con respecto a S (p=0,002). En S se encontraron las siguientes asociaciones negativas: IL-6 y AG n-3 docosahexaenoico (-0,37; p=0,013); TNF y AG n-3 eicosapentaenoico (-0,52; p<0,001) y docosahexaenoico (-0,94; p <0,001). Se encontraron asociaciones positivas entre IL-6 y AG saturado araquídico (0,73; p=0,01) e IL-1&#946; y AG saturado láurico (0,65; p=0,038) en CBCE, así como entre 12-HHT y AG n-6 linoleico (0,61; p=0,041) y 5-HETE y AG n-6 araquidónico (0,63; p=0,043) en ELB. En CBCE se encontraron las siguientes asociaciones negativas: 12-HHT y AG n-6 gamma-linolénico (-0,64; p=0,033); 13-HODE y AG n-6 linoleico (-0,72, p=0,013).Conclusión: la ingesta de AG n-3, n-6 y saturados influiría en el estado inflamatorio de la mucosa bucal al modular la producción de diferentes citoquinas y eicosanoides. Es necesario continuar investigando a fin de contribuir a la prevención y el diagnóstico precoz del cáncer bucal.OBJECTIVES: To analyze the relationship between dietary fatty acids (FA) and inflammatory salivary markers in oral lichenoid diseases (OLD) and oral squamous cell carcinoma (OSCC). METHODS: 31 persons bearing OLD (n=20) and OSCC (n=11), and 44 healthy volunteers (H) (40-80 years old in all groups) were included. A validated qualitative-quantitative food frequency questionnaire was employed. Data were processed using Interfood v.1.3. Mixed unstimulated saliva was obtained from each participant to analyze the following cytokines and eicosanoids: interleukins 1β (IL-1β), 6 (IL-6), tumor necrosis factor (TNF) (ELISA) and 12-hydroxyheptadecatrienoic acid (12-HHT), 13-hydroxyoctadecadienoic acid (13-HODE), 12 and 5 hydroxyeicosatetraenoic acid (12 and 5-HETE) (HPLC). Kruskal Wallis and Spearman’s coefficient tests were employed (p<0.05). RESULTS: IL-6 and TNF levels were higher in oral lesion patients than in H (p<0.001 and p=0.005, respectively). 13-HODE, 5 and 12-HETE were higher in OLD than in OSCC, and their levels were also higher in diseased than in H persons (p=0.002). The following negative associations were found in H: IL-6 and docosahexaenoic FA (-0.37; p=0.013); TNF and dietary n-3 eicosapentaenoic (-0.52; p<0.001) and docosahexaenoic FA (-0.94; p<0.001). Positive associations were found between IL-6 and arachidic FA (0.73; p=0.01) and IL-1β and lauric saturated FA (0.65; p=0.038) in OSCC, as well as between 12-HHT and n-6 linoleic FA (0.61; p=0.041), and 5-HETE and n-6 arachidonic FA (0.63; p=0.043) in OLD. The following negative associations were found in OSCC: 12-HHT and dietary n-6 gamma-linolenic FA (-0.64; p=0.033); 13-HODE and dietary n-6 linoleic FA (-0.72, p=0.013). CONCLUSIONS: The intake of n-3, n-6 and saturated FA appear to influence the inflammatory state of the oral mucosa by modulating the production of different cytokines and eicosanoids. Further research is necessary in order to contribute to oral cancer prevention and early diagnosis.Fil: Costantino, Evangelina. Universidad Nacional de Córdoba. Facultad de Odontología. Cátedra de Anatomía B; Argentina.Fil: Costantino, Evangelina. Universidad Nacional de Córdoba. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ciencias de la Salud; Argentina.Fil: Castell, Sofía Daiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina.Fil: Panico, René Luis. Universidad Nacional de Córdoba. Facultad de Odontología. Cátedra de Estomatología A; Argentina.Fil: Pascualini, María Eugenia. Universidad Nacional de Córdoba. Facultad de Ciencias Médicas. Cátedra de Biología Celular, Histología y Embriología; Argentina.Fil: Pascualini, María Eugenia, Pablo Gastón. Universidad Nacional de Córdoba. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ciencias de la Salud; Argentina.Fil: Pistoresi Palencia, María C. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina.Fil: Actis, Adriana Beatríz. Universidad Nacional de Córdoba. Facultad de Odontología. Secretaría de Ciencia y Técnica; Argentina.Fil: Actis, Adriana Beatriz. Universidad Nacional de Córdoba. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ciencias de la Salud; Argentina.Otras Ciencias de la Salu

Class-B CpG-ODN Formulated With a Nanostructure Induces Type I Interferons-Dependent and CD4+ T Cell-Independent CD8+ T-Cell Response Against Unconjugated Protein Antigen

There is a need for new vaccine adjuvant strategies that offer both vigorous antibody and T-cell mediated protection to combat difficult intracellular pathogens and cancer. To this aim, we formulated class-B synthetic oligodeoxynucleotide containing unmethylated cytosine-guanine motifs (CpG-ODN) with a nanostructure (Coa-ASC16 or coagel) formed by self-assembly of 6-0-ascorbyl palmitate ester. Our previous results demonstrated that mice immunized with ovalbumin (OVA) and CpG-ODN formulated with Coa-ASC16 (OVA/CpG-ODN/Coa-ASC16) elicited strong antibodies (IgG1 and IgG2a) and Th1/Th17 cellular responses without toxic systemic effects. These responses were superior to those induced by a solution of OVA with CpG-ODN or OVA/CpG-ODN formulated with aluminum salts. In this study, we investigated the capacity of this adjuvant strategy (CpG-ODN/Coa-ASC16) to elicit CD8+ T-cell response and some of the underlying cellular and molecular mechanisms involved in adaptive response. We also analyzed whether this adjuvant strategy allows a switch from an immunization scheme of three-doses to one of single-dose. Our results demonstrated that vaccination with OVA/CpG-ODN/Coa-ASC16 elicited an antigen-specific long-lasting humoral response and importantly-high quality CD8+ T-cell immunity with a single-dose immunization. Moreover, Coa-ASC16 promoted co-uptake of OVA and CpG-ODN by dendritic cells. The CD8+ T-cell response induced by OVA/CpG-ODN/Coa-ASC16 was dependent of type I interferons and independent of CD4+ T-cells, and showed polyfunctionality and efficiency against an intracellular pathogen. Furthermore, the cellular and humoral responses elicited by the nanostructured formulation were IL-6-independent. This system provides a simple and inexpensive adjuvant strategy with great potential for future rationally designed vaccines

Aging Impairs the Ability of Conventional Dendritic Cells to Cross-Prime CD8⁺ T Cells upon Stimulation with a TLR7 Ligand

<div><p>The aging process is accompanied by altered immune system functioning and an increased risk of infection. Dendritic cells (DCs) are antigen-presenting cells that play a key role in both adaptive and innate immunity, but how aging affects DCs and their influence on immunity has not been thoroughly established. Here we examined the function of conventional DCs (cDCs) in old mice after TLR7 stimulation, focusing on their ability to cross-prime CD8⁺ T cells. Using polyU, a synthetic ssRNA analog, as TLR7 ligand and OVA as an antigen (Ag) model, we found that cDCs from old mice have a poor ability to stimulate a CD8⁺ T cell-mediated cytotoxic response. cDCs from old mice exhibit alterations in Ag-processing machinery and TLR7 activation. Remarkably, CD8α⁺ cDCs from old mice have an impaired ability to activate naïve CD8⁺ T cells and, moreover, a lower capacity to mature and to process exogenous Ag. Taken together, our results suggest that immunosenescence impacts cDC function, affecting the activation of naïve CD8⁺ T cells and the generation of effector cytotoxic T cells.</p></div