RESUMO
BACKGROUND: Immunobiotic Lactobacillus jensenii TL2937 modulates porcine mononuclear phagocytes from Peyer's patches (PPMPs) and induces a differential production of pro- and anti-inflammatory cytokines in response to Toll-like receptor (TLR)-4 activation. In view of the important role played by phagocytosis in the activation of antigen presenting cells (APCs), the aim of the present work was to examine the interaction of TL2937 with porcine PPMPs focusing on phagocytosis. In addition, this study aimed to investigate whether the effects of L. jensenii TL2937 in porcine blood monocyte-derived dendritic cells (MoDCs) are similar to those found in PPMPs considering that MoDCs do not recapitulate all functions of mucosal APCs. RESULTS: Studies showed a high ability of porcine CD172a(+) PPMPs to phagocytose L. jensenii TL2937. Interestingly, our results also revealed a reduced capacity of the non-immunomodulatory L. plantarum TL2766 to be phagocytosed by those immune cells. Phagocytosis of L. jensenii TL2937 by porcine PPMPs was partially dependent on TLR2. In addition, we demonstrated that TL2937 strain was able to improve the expression of IL-1ß, IL-12 and IL-10 in immature MoDCs resembling the effect of this immunobiotic bacterium on PPMPs. Moreover, similarly to PPMPs those immunomodulatory effects were related to the higher capacity of TL2937 to be phagocytosed by immature MoDCs. CONCLUSIONS: Microbial recognition in APCs could be effectively mediated through ligand-receptor interactions that then mediate phagocytosis and signaling. For the immunobiotic strain TL2937, TLR2 has a partial role for its interaction with porcine APCs and it is necessary to investigate the role of other receptors. A challenge for future research will be advance in the full understanding of the molecular interactions of immunobiotic L. jensenii TL2937 with porcine APCs that will be crucial for the successful development of functional feeds for the porcine host. This study is a step in that direction.
Assuntos
Células Apresentadoras de Antígenos/imunologia , Células Dendríticas/imunologia , Imunomodulação , Mucosa Intestinal/imunologia , Lactobacillus johnsonii/imunologia , Monócitos/imunologia , Fagocitose , Animais , Células Cultivadas , Interleucina-10/metabolismo , Interleucina-12/metabolismo , Interleucina-1beta/metabolismo , Probióticos , Especificidade da Espécie , Suínos , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismoRESUMO
Radionuclides released and deposited because of the 2011 Fukushima Dai-ichi Nuclear Power Plant accident caused an increase in air dose rates in Fukushima Prefecture forests. Although an increase in air dose rates during rainfall was previously reported, the air dose rates in the Fukushima forests decreased during rainfall. This study aimed to develop a method to estimate rainfall-related changes in air dose rates, even in the absence of soil moisture data, in Namie-Town and Kawauchi-Village, Futaba-gun, Fukushima Prefecture. Moreover, we examined the relationship between preceding rainfall (Rw) and soil moisture content. The air dose rate was estimated by calculating the Rw in Namie-Town from May to July 2020. We found that the air dose rates decreased with increasing soil moisture content. The soil moisture content was estimated from Rw by combining short-term and long-term effective rainfall using half-live values of 2 h and 7 d and considering the hysteresis of water absorption and drainage processes. Furthermore, the soil moisture content and air dose rate estimations showed a good agreement with coefficient of determination (R2) scores >0.70 and >0.65, respectively. The same method was tested to estimate the air dose rates in Kawauchi-Village from May to July 2019. At the Kawauchi site, variation in estimated value is relatively large due to the presence of water repellency in dry conditions, and the amount of 137Cs inventory was low, so estimating air dose from rainfall remained a challenge. In conclusion, rainfall data were successfully used to estimate soil moisture and air dose rates in areas with high 137Cs inventories. This leads to the possibility of removing the influence of rainfall on measured air dose rate data and could contribute to the improvement of methods currently used to estimate the external air dose rates for humans, animals, and terrestrial forest plants.
Assuntos
Acidente Nuclear de Fukushima , Monitoramento de Radiação , Poluentes Radioativos do Solo , Humanos , Animais , Monitoramento de Radiação/métodos , Solo , Poluentes Radioativos do Solo/análise , Florestas , Radioisótopos de Césio/análise , Água , JapãoRESUMO
Emerging threats of antimicrobial resistance necessitate the exploration of effective alternatives for healthy livestock growth strategies. 'Immunosynbiotics', a combination of immunoregulatory probiotics and prebiotics with synergistic effects when used together in feed, would be one of the most promising candidates. Lactobacilli are normal residents of the gastrointestinal tract of pigs, and many of them are able to exert beneficial immunoregulatory properties. On the other hand, wakame (Undaria pinnafida), an edible seaweed, has the potential to be used as an immunoregulatory prebiotic when added to livestock feed. Therefore, in order to develop a novel immunosynbiotic, we isolated and characterized immunoregulatory lactobacilli with the ability to utilize wakame. Following a month-long in vivo wakame feeding trial in 8-week-old Landrace pigs (n = 6), sections of intestinal mucous membrane were processed for bacteriological culture and followed by identification of pure colonies by 16S rRNA sequence. Each isolate was characterized in vitro in terms of their ability to assimilate to the wakame and to differentially modulate the expression of interleukin-6 (IL-6) and interferon beta (IFN-ß) in the porcine intestinal epithelial (PIE) cells triggered by Toll-like receptor (TLR)-4 and TLR-3 activation, respectively. We demonstrated that feeding wakame to pigs significantly increased the lactobacilli population in the small intestine. We established a wakame-component adjusted culture media that allowed the isolation and characterization of a total of 128 Lactobacilli salivarius colonies from the gut of wakame-fed pigs. Interestingly, several L. salivarius isolates showed both high wakame assimilation ability and immunomodulatory capacities. Among the wakame assimilating isolates, L. salivarius FFIG71 showed a significantly higher capacity to upregulate the IL-6 expression, and L. salivarius FFIG131 showed significantly higher capacity to upregulate the IFN-ß expression; these could be used as immunobiotic strains in combination with wakame for the development of novel immunologically active feeds for pigs.
RESUMO
Peptidoglycan recognition proteins (PGLYRPs) are a family of pattern recognition receptors (PRRs) that are able to induce innate immune responses through their binding to peptidoglycan (PGN), lipopolysaccharide, or lipoteichoic acid, or by interacting with other PRR-ligands. Recently, progress has been made in understanding the immunobiology of PGLYRPs in human and mice, however, their functions in livestock animals have been less explored. In this study, we characterized the expression patterns of PGLYRPs in porcine intestinal epithelial (PIE) cells and antigen-presenting cells (APCs) and their modulation by the interactions of host cells with PRR-ligands and non-viable immunomodulatory probiotics referred to as paraimmunobiotics. We demonstrated that PGLYRP-1, -2, -3, and -4 are expressed in PIE cells and APCs from Peyer's patches, being PGLYPR-3 and -4 levels higher than PGLYRP-1 and -2. We also showed that PGLYRPs expression in APCs and PIE cells can be modulated by different PRR agonists. By using knockdown PIE cells for TLR2, TLR4, NOD1, and NOD2, or the four PGLYRPs, we demonstrated that PGLYRPs expressions would be required for activation and functioning of TLR2, TLR4, NOD1, and NOD2 in porcine epitheliocytes, but PGLYRPs activation would be independent of those PRR expressions. Importantly, we reported for the first time that PGLYRPs expression can be differentially modulated by paraimmunobiotic bifidobacteria in a strain-dependent manner. These results provide evidence for the use of paraimmunobiotic bifidobacteria as an alternative for the improvement of resistance to intestinal infections or as therapeutic tools for the reduction of the severity of inflammatory damage in diseases in which a role of PGLYRPs-microbe interaction has been demonstrated.
Assuntos
Bifidobacterium/fisiologia , Proteínas de Transporte/metabolismo , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Suínos/imunologia , Suínos/microbiologia , Animais , Células Apresentadoras de Antígenos/imunologia , Nódulos Linfáticos Agregados/citologia , Nódulos Linfáticos Agregados/imunologia , Receptores de Reconhecimento de Padrão/metabolismo , Baço/citologia , Baço/imunologiaRESUMO
Previous studies demonstrated that the extracellular polysaccharides (EPSs) produced by Lactobacillus delbrueckii OLL1073R-1 (LDR-1) improve antiviral immunity, especially in the systemic and respiratory compartments. However, it was not studied before whether those EPSs are able to beneficially modulate intestinal antiviral immunity. In addition, LDR-1-host interaction has been evaluated mainly with immune cells while its interaction with intestinal epithelial cells (IECs) was not addressed before. In this work, we investigated the capacity of EPSs from LDR-1 to modulate the response of porcine IECs (PIE cells) to the stimulation with the Toll-like receptor (TLR)-3 agonist poly(I:C) and the role of TLR2, TLR4, and TLR negative regulators in the immunoregulatory effect. We showed that innate immune response triggered by TLR3 activation in porcine IECs was differentially modulated by EPS from LDR-1. EPSs treatment induced an increment in the expression of interferon (IFN)-α and IFN-ß in PIE cells after the stimulation with poly(I:C) as well as the expression of the antiviral factors MxA and RNase L. Those effects were related to the reduced expression of A20 in EPS-treated PIE cells. EPS from LDR-1 was also able to reduce the expression of IL-6 and proinflammatory chemokines. Although further in vivo studies are needed, our results suggest that these EPSs or a yogurt fermented with LDR-1 have potential to improve intestinal innate antiviral response and protect against intestinal viruses.
Assuntos
Células Epiteliais/efeitos dos fármacos , Imunidade Inata/efeitos dos fármacos , Interferon beta/biossíntese , Mucosa Intestinal/citologia , Lactobacillus delbrueckii/imunologia , Polissacarídeos Bacterianos/farmacologia , Sus scrofa/imunologia , Animais , Células Cultivadas , Citocinas/biossíntese , Citocinas/genética , Células Epiteliais/imunologia , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Interferon beta/genética , Lactobacillus delbrueckii/química , Poli I-C/farmacologia , Polissacarídeos Bacterianos/isolamento & purificação , Transdução de Sinais , Suínos , Doenças dos Suínos/imunologia , Receptor 2 Toll-Like/imunologia , Receptor 3 Toll-Like/imunologia , Receptor 4 Toll-Like/imunologia , Viroses/imunologia , Viroses/veterináriaRESUMO
The genome of the immunomodulatory strain Lactobacillus jensenii TL2937 is described here. The draft genome has a total length of 1,678,416 bp, a G+C content of 34.3%, and 1,470 predicted protein-coding sequences. The genome information will be useful for gaining insight into the immunomodulatory properties of the TL2937 strain in the porcine host.
RESUMO
Lactobacillus rhamnosus CRL1505 and Lactobacillus plantarum CRL1506 are immunobiotic strains able to increase protection against viral intestinal infections as demonstrated in animal models and humans. To gain insight into the host-immunobiotic interaction, the transcriptomic response of porcine intestinal epithelial (PIE) cells to the challenge with viral molecular associated pattern poly(I:C) and the changes in the transcriptomic profile induced by the immunobiotics strains CRL1505 and CRL1506 were investigated in this work. By using microarray technology and reverse transcription PCR, we obtained a global overview of the immune genes involved in the innate antiviral immune response in PIE cells. Stimulation of PIE cells with poly(I:C) significantly increased the expression of IFN-α and IFN-ß, several interferon-stimulated genes, cytokines, chemokines, adhesion molecules, and genes involved in prostaglandin biosynthesis. It was also determined that lactobacilli differently modulated immune gene expression in poly(I:C)-challenged PIE cells. Most notable changes were found in antiviral factors (IFN-α, IFN-ß, NPLR3, OAS1, OASL, MX2, and RNASEL) and cytokines/chemokines (IL-1ß, IL-6, CCL4, CCL5, and CXCL10) that were significantly increased in lactobacilli-treated PIE cells. Immunobiotics reduced the expression of IL-15 and RAE1 genes that mediate poly(I:C) inflammatory damage. In addition, lactobacilli treatments increased the expression PLA2G4A, PTGES, and PTGS2 that are involved in prostaglandin E2 biosynthesis. L. rhamnosus CRL1505 and L. plantarum CRL1506 showed quantitative and qualitative differences in their capacities to modulate the innate antiviral immune response in PIE cells, which would explain the higher capacity of the CRL1505 strain when compared to CRL1506 to protect against viral infection and inflammatory damage in vivo. These results provided valuable information for the deeper understanding of the host-immunobiotic interaction and their effect on antiviral immunity. The comprehensive transcriptomic analyses successfully identified a group of genes (IFN-ß, RIG1, RNASEL, MX2, A20, IL27, CXCL5, CCL4, PTGES, and PTGER4), which can be used as prospective biomarkers for the screening of new antiviral immunobiotics in PIE cells and for the development of novel functional food and feeds, which may help to prevent viral infections.