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1.
Transplant Proc ; 51(7): 2461-2465, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31474299

RESUMEN

Carbapenemase-producing Enterobacteriacea (CPE) cause serious and life-threatening infections. They are resistant to carbapenems and many other classes of commonly used antimicrobial agents; therefore, managing infections caused by them poses a substantial challenge in clinical practice. They can also cause morbidity and mortality in patients with liver transplant. A retrospective analysis of CPE culture-positive patients with a history of liver transplant can help to examine the epidemiology and microbiology of these bacteria, as well as gain information on the possible infection sources, susceptibility patterns, and expected mortality in infected populations. In addition, study of these bacteria could help formulate a consensus on the appropriate use of empirical and directed antibiotic therapy, which can effectively reduce infections in these patients. We reviewed the medical records of 142 subjects who underwent liver transplantation at Ankara University Hospital, a 1900-bed tertiary care university hospital, in Ankara, Turkey, between January 2014 and August 2018. Patients showing signs of infection with culture positivity for CPE-producing organisms were included from the study. Statistical analysis was performed and a value of P < .05 is considered statistically significant. In most cases, the source of infection was the abdomen. Klebsiella species was also predominant in these cases. Model for End-Stage Liver Disease scores and length of hospital stay were higher and statistically significant when compared to patients who were CPE negative. Mortality was highest in the CPE-positive group. Infection is the most important cause of mortality and morbidity after liver transplantation and increases the cost of treatment. Regarding the culture sensitivity patterns and resistance mode, empirical therapy with carbapenems does not produce a solid result. The high mortality observed with these infections reflects very limited therapeutic options.


Asunto(s)
Farmacorresistencia Microbiana , Infecciones por Enterobacteriaceae/epidemiología , Trasplante de Hígado/efectos adversos , Complicaciones Posoperatorias/microbiología , Adulto , Anciano , Proteínas Bacterianas , Infecciones por Enterobacteriaceae/inmunología , Femenino , Humanos , Huésped Inmunocomprometido , Incidencia , Trasplante de Hígado/mortalidad , Masculino , Persona de Mediana Edad , Complicaciones Posoperatorias/inmunología , Estudios Retrospectivos , Turquia , beta-Lactamasas
2.
Fish Shellfish Immunol ; 93: 766-780, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31421241

RESUMEN

Japanese flounder (Paralichthys olivaceus) is an important economic fish species farmed in China and other countries. It is susceptible to infection by Edwardsiella tarda, a severe fish pathogen with a broad host range. In this study, we employed high-throughput deep sequencing technology to identify, in a global scale, flounder kidney microRNAs (miRNAs) induced by E. tarda at different stages of infection. Differentially expressed miRNAs (DEmiRNAs) and mRNAs (DEmRNAs) exhibiting significantly altered expression levels before and after E. tarda infection were examined. A total of 96 DEmiRNAs were identified, for which 2779 target genes were predicted. Eighty-seven miRNA-mRNA pairs, involving 29 DEmiRNAs and 86 DEmRNAs, showed negative correlations in their expression patterns. GO and KEGG enrichment analysis revealed that the putative target genes of the DEmiRNAs were associated with diverse biological processes, cellular components, and molecular functions. One of the DEmiRNAs, pol-miR-182-5p, was demonstrated to regulate sphingosine-1-phosphate receptor 1 (PoS1PR1) negatively in a manner that depended on the specific interaction between the seed sequence of pol-miR-182-5p and the 3'-UTR of PoS1PR1. Overexpression of pol-miR-182-5p in flounder cells promoted apoptosis and inhibited cellular viability. Knockdown of PoS1PR1 in flounder enhanced E. tarda invasion and dissemination in fish tissues. These results provide new insights into miRNA-mediated anti-bacterial immunity in flounder.


Asunto(s)
Enfermedades de los Peces/inmunología , Peces Planos/genética , Regulación de la Expresión Génica/inmunología , MicroARNs/genética , Secuencia de Aminoácidos , Animales , Edwardsiella tarda/fisiología , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/veterinaria , Peces Planos/inmunología , Perfilación de la Expresión Génica/veterinaria , Riñón/inmunología , Riñón/metabolismo , MicroARNs/inmunología , Alineación de Secuencia , Factores de Tiempo
3.
Fish Shellfish Immunol ; 93: 823-831, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31422181

RESUMEN

Calreticulin (CRT) is a highly conserved and multi-functional protein with diverse localizations. CRT has lectin-like properties and possesses important immunological activities in mammalian. In teleost, very limited studies on CRT immunologic function have been documented. In the present study, a CRT homologue (SsCRT) was cloned, identified and characterized from black rockfish, Sebastes schlegeli, an important aquaculture species in East Asia. The full length of SsCRT cDNA is 2180 bp and encoded a polypeptide of 425 amino acids. SsCRT contains a signal peptide, three distinct structural and functional domains (N-, P- and C-domains), and an endoplasmic reticulum (ER) retrieval signal sequence (KDEL). The deduced amino acid sequence of SsCRT shares 89-92% overall sequence identities with the CRT proteins of several fish species. SsCRT was distributed ubiquitously in all the detected tissues and was highly expressed in the spleen, muscle and liver. After the infection of fish extracellular bacterial pathogen Vibrio anguillarum and intracellular bacterial pathogen Edwardsiella tarda, the mRNA transcripts of SsCRT in spleen, liver, and head kidney were significantly up-regulated. The expression patterns were time-dependent and tissue-dependent. Recombinant SsCRT (rSsCRT) exhibited apparent binding activities against different bacteria and PAMPs. In vivo studies showed that the expressions of multiple immune-related genes such as TNF13B, IL-1ß, IL-8, SAA, Hsp70, and ISG15 in head kidney were significantly enhanced when black rockfish were treated with rSsCRT. Furthermore, rSsCRT reduced pathogen dissemination and replication in fish kidney and spleen. These results indicated that SsCRT served as an immune receptor to recognize and eliminate the invading pathogens, which played a vital role in the immune response of Sebastes schlegeli. These findings provide new insights into understanding the roles of CRT proteins in immune response and pathogen infection in teleost.


Asunto(s)
Calreticulina/genética , Calreticulina/inmunología , Enfermedades de los Peces/inmunología , Peces/genética , Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Calreticulina/química , Edwardsiella tarda/fisiología , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/veterinaria , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Patrón Molecular Asociado a Patógenos/farmacología , Perciformes/genética , Perciformes/inmunología , Filogenia , Alineación de Secuencia/veterinaria , Vibrio/fisiología , Vibriosis/inmunología , Vibriosis/veterinaria
4.
Fish Shellfish Immunol ; 93: 871-878, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31400510

RESUMEN

Edwardsiella piscicida is an important pathogen that infects a wide range of hosts, from fish to human. Its infection leads to extensive losses in a diverse array of commercially important fish, like Japanese flounder, turbot, and tilapia. During the infection, type III secretion system (T3SS) and type VI secretion system (T6SS) of E. piscicida play significant roles, but how T3SS and T6SS cooperatively contribute to its virulence is still unknown. In this study, we first examined the roles of T3SS and T6SS in different processes during E. piscicida infection of host cells, and revealed that T3SS of E. piscicida is responsible for promoting bacterial invasion, the following intracellular replication and inducing cell death in host cells, while T6SS restrains E. piscicida intracellular replication and cell death in J774A.1 cells, which suggested that T3SS and T6SS antagonistically concert E. piscicida infection. Furthermore, we found an significant decrease in transcription level of IL-1ß in zebrafish kidney infected with T3SS mutant and an drastically increase in transcription level of TNF- α infected with T6SS mutant when compared with the wild-type. Interestingly, both T3SS and T6SS mutants showed significant attenuated virulence in the zebrafish infection model when compared with the wild-type. Finally, considering the cooperative role of T3SS and T6SS, we generated a mutant strain WEDΔT6SS based on the existing live attenuated vaccine (LAV) WED which showed improved vaccine safety and comparable immune protection. Therefore, WEDΔT6SS could be used as an optimized LAV in the future. Taken together, this work suggested a bilateral role of T3SS and T6SS which respectively act as spear and shield during E. piscicida infection, together contribute to E. piscicida virulence.


Asunto(s)
Vacunas Bacterianas/farmacología , Edwardsiella/patogenicidad , Enfermedades de los Peces/inmunología , Sistemas de Secreción Tipo III/genética , Sistemas de Secreción Tipo VI/genética , Pez Cebra/inmunología , Animales , Línea Celular Tumoral , Edwardsiella/genética , Edwardsiella/fisiología , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/veterinaria , Células HeLa , Interacciones Huésped-Patógeno , Humanos , Ratones , Sistemas de Secreción Tipo III/metabolismo , Sistemas de Secreción Tipo VI/metabolismo , Vacunas Atenuadas/farmacología , Virulencia
5.
Nature ; 571(7766): 565-569, 2019 07.
Artículo en Inglés | MEDLINE | ID: mdl-31316206

RESUMEN

Parkinson's disease is a neurodegenerative disorder with motor symptoms linked to the loss of dopaminergic neurons in the substantia nigra compacta. Although the mechanisms that trigger the loss of dopaminergic neurons are unclear, mitochondrial dysfunction and inflammation are thought to have key roles1,2. An early-onset form of Parkinson's disease is associated with mutations in the PINK1 kinase and PRKN ubiquitin ligase genes3. PINK1 and Parkin (encoded by PRKN) are involved in the clearance of damaged mitochondria in cultured cells4, but recent evidence obtained using knockout and knockin mouse models have led to contradictory results regarding the contributions of PINK1 and Parkin to mitophagy in vivo5-8. It has previously been shown that PINK1 and Parkin have a key role in adaptive immunity by repressing presentation of mitochondrial antigens9, which suggests that autoimmune mechanisms participate in the aetiology of Parkinson's disease. Here we show that intestinal infection with Gram-negative bacteria in Pink1-/- mice engages mitochondrial antigen presentation and autoimmune mechanisms that elicit the establishment of cytotoxic mitochondria-specific CD8+ T cells in the periphery and in the brain. Notably, these mice show a sharp decrease in the density of dopaminergic axonal varicosities in the striatum and are affected by motor impairment that is reversed after treatment with L-DOPA. These data support the idea that PINK1 is a repressor of the immune system, and provide a pathophysiological model in which intestinal infection acts as a triggering event in Parkinson's disease, which highlights the relevance of the gut-brain axis in the disease10.


Asunto(s)
Infecciones por Enterobacteriaceae/microbiología , Infecciones por Enterobacteriaceae/fisiopatología , Intestinos/microbiología , Enfermedad de Parkinson/genética , Enfermedad de Parkinson/microbiología , Proteínas Quinasas/deficiencia , Proteínas Quinasas/genética , Animales , Presentación de Antígeno/inmunología , Autoantígenos/inmunología , Axones/patología , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/patología , Citrobacter rodentium/inmunología , Citrobacter rodentium/patogenicidad , Modelos Animales de Enfermedad , Neuronas Dopaminérgicas/inmunología , Neuronas Dopaminérgicas/patología , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/patología , Femenino , Intestinos/inmunología , Intestinos/patología , Levodopa/uso terapéutico , Masculino , Ratones , Mitocondrias/inmunología , Mitocondrias/patología , Neostriado/inmunología , Neostriado/microbiología , Neostriado/patología , Neostriado/fisiopatología , Enfermedad de Parkinson/tratamiento farmacológico , Enfermedad de Parkinson/fisiopatología , Proteínas Quinasas/inmunología , Ubiquitina-Proteína Ligasas/deficiencia , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/inmunología
6.
Fish Shellfish Immunol ; 93: 208-215, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31306760

RESUMEN

Cathepsin Z (CTSZ) is a lysosomal cysteine protease that is known to be involved in the maintenance of homeostasis and the biological mechanisms of immune cells. In this study, we have confirmed the tissue specific expression of the cathepsin Z (PmCTSZ) gene in Pagrus major, and confirmed its biological function after producing recombinant protein using Escherichia coli (E. coli). Multiple sequence alignment analysis revealed that the active site of the cysteine proteases and three N-glycosylation sites of the deduced protein sequence were highly conserved among all of the organisms. Phylogenetic analysis revealed that PmCTSZ was included in the clusters of CTSZ and the cysteine proteases of other bony fish and is most closely related to Japanese flounder CTSZ. PmCTSZ was distributed in all of the tissues from healthy red sea bream that were used in the experiment and was most abundantly found in the spleen and gill. Analysis of mRNA expression after bacterial (Edwardsiella piscicida: E. piscicida and Streptococcus iniae: S. iniae) or viral (red seabream iridovirus: RSIV) challenge showed significant gene expression regulation in immune-related tissues, but they maintained relatively normal levels of expression. We produced recombinant PmCTSZ (rPmCTSZ) using an E. coli expression system and confirmed the biological function of extracellular rPmCTSZ in vitro. We found that bacterial proliferation was significantly inhibited by rPmCTSZ, and the leukocytes of red sea bream also induced apoptosis and viability reduction.


Asunto(s)
Catepsina Z/genética , Catepsina Z/inmunología , Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Dorada/genética , Dorada/inmunología , Secuencia de Aminoácidos , Animales , Catepsina Z/química , Infecciones por Virus ADN/inmunología , Infecciones por Virus ADN/veterinaria , Edwardsiella/fisiología , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/veterinaria , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Iridoviridae/fisiología , Filogenia , Alineación de Secuencia/veterinaria , Infecciones Estreptocócicas/inmunología , Infecciones Estreptocócicas/veterinaria , Streptococcus iniae/fisiología
7.
Fish Shellfish Immunol ; 92: 913-924, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31306761

RESUMEN

A feeding trial was performed to compare the effects of five ethanol herbal extracts (bhumi amla, Phyllanthus amarus Schum and Thonn [Pa]; guava, Psidium guajava L. [Pg]; sensitive plant, Mimosa pudica L. [Mp]; neem, Azadirachta indica A. Juss [Ai] and asthma plant, Euphorbia hirta L. [Eh]) on the immune response and disease resistance against Edwardsiella ictaluri infection of striped catfish (Pangasianodon hypophthalmus). Fish were fed diets supplemented with two doses of each plant extract (0% [basal diet], 0.4% Eh [Eh0.4], 2.0% Eh [Eh2.0], 0.2% Pa [Pa0.2], 1.0% Pa [Pa1.0], 0.2% Pg [Pg0.2], 1.0% Pg [Pg1.0], 0.4% Mp [Mp0.4], 2.0% Mp [Mp2.0], 0.4% Ai [Ai0.4], 2.0% Ai [Ai2.0]) for 8 weeks. Results showed that hematological parameters (total red blood cells, white blood cells, lymphocytes, monocytes, and neutrophils) of fish fed extract-based diets were significantly higher than in those fed the control diet (p < 0.05) after 4 and 8 weeks. Plasma lysozyme activity increased in fish whose diets contained both doses of Eh (p < 0.05) in week 4 (W4), whereas lysozyme activity increased in fish fed 0.2% Pa and Pg, and 2.0% Ai and Eh (p < 0.05) in week 8 (W8). The lysozyme levels in skin mucus did not significantly differ between treatments (p > 0.05) in W4 and after the bacterial challenge test. At the end of the feeding trial, levels of ACH50 significantly increased in most of extract groups compared to the control group (p < 0.05). Total immunoglobulin increased considerably in both the plasma and skin mucus of fish fed extract-supplemented diets after 8 weeks. In addition, dietary supplementation with Pg, Mp, Pa0.2, Eh2.0, and Ai0.4 for 8 weeks considerably reduced the cumulative mortality against E. ictaluri infection in striped catfish. The results suggest that plant extracts possibly modulate the striped catfish immune response in a time and dose dependent manner. Specifically, diets enriched with extracts of P. guajava at 0.2 and 1.0%, or M. pudica at 2.0% for 8 weeks, have great potential for improving striped catfish health by enhancing the immune system and reducing mortality against bacterial challenges.


Asunto(s)
Inmunidad Adaptativa/efectos de los fármacos , Bagres/inmunología , Resistencia a la Enfermedad/efectos de los fármacos , Inmunidad Innata/efectos de los fármacos , Extractos Vegetales/metabolismo , Alimentación Animal/análisis , Animales , Dieta/veterinaria , Suplementos Dietéticos/análisis , Relación Dosis-Respuesta a Droga , Edwardsiella ictaluri/fisiología , Infecciones por Enterobacteriaceae/inmunología , Enfermedades de los Peces/inmunología , Extractos Vegetales/administración & dosificación , Distribución Aleatoria
8.
J Dairy Sci ; 102(9): 8352-8366, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31255273

RESUMEN

Calf preweaning morbidity and mortality risks have been reported as high in several countries, with average values approximating 35 and 7%, respectively. However, limited data are available for calf morbidity and mortality risks on Australian dairy farms. The aims of this study were (1) to investigate current calf management practices on dairy farms in Australia and their association with herd-level morbidity and mortality using a questionnaire-based, cross-sectional study; and (2) to estimate the prevalence of common enteropathogens causing diarrhea, the failure of passive transfer of immunity, and poor colostrum quality in a sample of Australian dairy farms. We analyzed 106 completed questionnaires and samples from 23 farms (202 fecal, 253 calf serum, and 221 colostrum samples). Morbidity and mortality risks reported by farmers in preweaned heifers were 23.8 and 5.6%, respectively. These risks were above the Australian dairy industry targets in 75.5 and 66.7% of respondents. The zoonotic pathogens Cryptosporidium spp. and Salmonella spp. were the most prevalent enteropathogens, with a true prevalence of 40.9 and 25.2%, respectively. Salmonella O-group D was present in 67.9% of Salmonella-positive samples, followed by O-groups B (17.9%) and C (10.7%). Failure of transfer of passive immunity (IgG <10 g/L) was observed in 41.9% of calves (mean herd-level prevalence of 36.2%), and only 19.5% of colostrum samples met the standards for immunoglobulin content and microbiological quality. Collectively, these data indicate that there is still considerable room for improvement in calf-rearing practices on Australian dairy farms, particularly with regard to colostrum management and feeding hygiene.


Asunto(s)
Enfermedades de los Bovinos/inmunología , Calostro , Industria Lechera , Infecciones por Enterobacteriaceae/veterinaria , Animales , Animales Recién Nacidos , Australia , Bovinos , Enfermedades de los Bovinos/epidemiología , Calostro/inmunología , Estudios Transversales , Infecciones por Enterobacteriaceae/epidemiología , Infecciones por Enterobacteriaceae/inmunología , Agricultores , Granjas , Heces , Femenino , Higiene , Embarazo , Prevalencia
9.
Fish Shellfish Immunol ; 93: 55-65, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31319204

RESUMEN

Interleukin-2 (IL-2) is mainly produced by CD4+ T helper lymphocytes, which is an important immunomodulatory cytokine that primarily promotes activation, proliferation and differentiation of T cells. In the present study, flounder (Paralichthys olivaceus) interleukin 2 homologue (poIL-2) was identified for the first time, and its expression patterns were characterized in healthy, virus- or bacteria-infected flounder. The full-length cDNA sequences of poIL-2 was 989 bp with an open reading frame of 423 bp coding a polypeptide of 140 amino acids (aa). The deduced aa sequences shared low similarities (<53%) with other known fish IL-2s. Multiple alignment of aa sequences revealed that poIL-2 own the classical IL-2 family signature of "C-X(3)-EL-X(2)-(T/V)-(V/M/L)-(K/T/R)-X-EC" and "DS-X-(F/L)Y(A/T/S)P". In healthy flounder, IL-2 mRNA was highly expressed in PBLs, spleen and hindgut, and moderately expressed in gill, trunk kidney and stomach. PHA, LPS and Con-A could effectively induce poIL-2 expression in primary cultured peripheral blood leukocytes in vitro. poIL-2 transcripts were significantly up-regulated in spleen, kidney, gill and hindgut post infections with Edwardsiella tarda and Hirame novirhabdovirus (HIRRV). The eukaryotic expression vector encoding poIL-2 (pcIL-2) was constructed and intramuscularly injected, which could be successfully expressed in flounders and induced significantly higher expressions of six immune related genes including poIL-2, ß-defensin, CD4-1, CD8α, IFN-γ and TNF-α compared with the injection with control plasmid. Moreover, pretreatment with pcIL-2 could markedly increase the survival rate of flounder challenged with HIRRV. Our results demonstrated that poIL-2 plays an important role in the induction of immune responses and immune defense against bacterial and virus infection, which indicated its potential use as an immunopotentiator to prevent diseases in flounder.


Asunto(s)
Enfermedades de los Peces/inmunología , Peces Planos/genética , Peces Planos/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Interleucina-2/genética , Interleucina-2/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Edwardsiella tarda/fisiología , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/veterinaria , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Interleucina-2/química , Novirhabdovirus/fisiología , Filogenia , Infecciones por Rhabdoviridae/inmunología , Infecciones por Rhabdoviridae/veterinaria , Alineación de Secuencia/veterinaria
10.
Fish Shellfish Immunol ; 92: 356-366, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31200074

RESUMEN

Glutathione S-transferases (GSTs) are essential enzymes for the bioactivation of xenobiotics through the conjugation of the thiol group of glutathione (GSH). In this study, a kappa class of GST was identified from the big belly seahorse (Hippocampus abdominalis) (HaGSTκ1) and its biochemical and functional properties were analyzed. HaGSTκ1 has 231 amino acids encoded by a 696 bp open reading frame (ORF). The protein has a predicted molecular mass of 26.04 kDa and theoretical isoelectric point (pI) of 8.28. It comprised a thioredoxin domain, disulfide bond formation protein A (DsbA) general fold, and Ser15 catalytic site as well as GSH-binding and polypeptide-binding sites. Phylogenetic analysis revealed that HaGSTκ1 is closely clustered with the kappa class of GSTs from teleost fishes. The recombinant (rHaGSTκ1) protein exhibited activity toward 1-chloro-2,4-dinitrobenzene (CDNB), 4-nitrobenzyl (4-NBC), and 4-nitrophenethyl bromide (4-NPB) but not 1,2-dichloro-4-nitrobenzene (DCNB). The optimum pH and temperature were 8 and 30 °C, respectively, for the catalysis of CDNB and the universal substrate of GSTs. The rHaGSTκ1 activity was efficiently inhibited in the presence of Cibacron blue (CB) as compared with hematin. Most prominent expression of HaGSTκ1 was observed in the liver and kidney among the fourteen different tissues of normal seahorse. After challenge with lipopolysaccharide (LPS), polyinosinic-polycytidylic (poly I:C), gram-negative Edwardsiella tarda, and gram-positive Streptococcus iniae, HaGSTκ1 expression was significantly modulated in the liver and blood tissues. Altogether, our study proposes the plausible important role of HaGSTκ1 in innate immunity and detoxification of harmful xenobiotics.


Asunto(s)
Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Glutatión Transferasa/genética , Glutatión Transferasa/inmunología , Inmunidad Innata/genética , Smegmamorpha/genética , Smegmamorpha/inmunología , Animales , Edwardsiella tarda/fisiología , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/veterinaria , Femenino , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Glutatión Transferasa/química , Lipopolisacáridos/farmacología , Masculino , Conformación Molecular , Filogenia , Poli I-C/farmacología , Infecciones Estreptocócicas/inmunología , Infecciones Estreptocócicas/veterinaria , Streptococcus iniae/fisiología
11.
Fish Shellfish Immunol ; 92: 256-264, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31200076

RESUMEN

NK-lysin (NKL) is a cationic host defense peptide that plays an important role in host immune responses against various pathogens. However, the immunomodulatory activity of NKL in fishes is rarely investigated. In this study, we characterized a cDNA sequence encoding an NK-lysin homolog (BpNKL) from the fish, mudskipper (Boleophthalmus pectinirostris). Sequence analysis revealed that BpNKL is most closely related to tiger puffer (Takifugu rubripes) NKL. BpNKL transcript was detected in all the tested tissues, with the highest level in the gill, followed by the spleen and kidney. Upon Edwardsiella tarda infection, the mRNA expression of BpNKL in the mudskipper was significantly upregulated in the spleen, kidney, and gill. A shortened peptide derived from BpNKL, BpNKLP40, was then chemically synthesized and its biological functions were investigated. BpNKLP40 exhibited a direct antibacterial activity against some Gram-negative bacteria, including E. tarda, Vibrio parahaemolyticus, Vibrio alginolyticus, and Vibrio harveyi, and induced hydrolysis of E. tarda genomic DNA. Intraperitoneal injection of 1.0 µg/g BpNKLP40 significantly improved the survival of mudskipper following E. tarda infection and reduced the bacterial burden in tissues and blood. Moreover, 1.0 µg/ml BpNKLP40 treatment had an enhanced effect on the intracellular killing of E. tarda by monocytes/macrophages (MO/MФ) as well as on the mRNA expression of pro-inflammatory cytokines in MO/MФ. In conclusion, our study reveals that BpNKL plays a role against E. tarda infection in the mudskipper by not only directly killing bacteria but also through an immunomodulatory activity on MO/MФ.


Asunto(s)
Enfermedades de los Peces/inmunología , Peces/genética , Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Proteolípidos/genética , Proteolípidos/inmunología , Secuencia de Aminoácidos , Animales , Antiinfecciosos/farmacología , Edwardsiella tarda/fisiología , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/veterinaria , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Factores Inmunológicos/farmacología , Macrófagos/inmunología , Monocitos/inmunología , Filogenia , Proteolípidos/química , Alineación de Secuencia/veterinaria , Vibrio/fisiología , Vibriosis/inmunología , Vibriosis/veterinaria
12.
Immunity ; 51(1): 77-89.e6, 2019 07 16.
Artículo en Inglés | MEDLINE | ID: mdl-31229354

RESUMEN

T helper 17 (Th17) cells are pathogenic in many inflammatory diseases, but also support the integrity of the intestinal barrier in a non-inflammatory manner. It is unclear what distinguishes inflammatory Th17 cells elicited by pathogens and tissue-resident homeostatic Th17 cells elicited by commensals. Here, we compared the characteristics of Th17 cells differentiating in response to commensal bacteria (SFB) to those differentiating in response to a pathogen (Citrobacter rodentium). Homeostatic Th17 cells exhibited little plasticity towards expression of inflammatory cytokines, were characterized by a metabolism typical of quiescent or memory T cells, and did not participate in inflammatory processes. In contrast, infection-induced Th17 cells showed extensive plasticity towards pro-inflammatory cytokines, disseminated widely into the periphery, and engaged aerobic glycolysis in addition to oxidative phosphorylation typical for inflammatory effector cells. These findings will help ensure that future therapies directed against inflammatory Th17 cells do not inadvertently damage the resident gut population.


Asunto(s)
Citrobacter rodentium/inmunología , Infecciones por Enterobacteriaceae/inmunología , Microbioma Gastrointestinal/inmunología , Intestinos/inmunología , Células Th17/inmunología , Animales , Plasticidad de la Célula , Células Cultivadas , Citocinas/metabolismo , Glucólisis , Homeostasis , Memoria Inmunológica , Inflamación , Ratones , Ratones Transgénicos
13.
PLoS Pathog ; 15(6): e1007898, 2019 06.
Artículo en Inglés | MEDLINE | ID: mdl-31251784

RESUMEN

Attaching/Effacing (A/E) bacteria include human pathogens enteropathogenic Escherichia coli (EPEC), enterohemorrhagic E. coli (EHEC), and their murine equivalent Citrobacter rodentium (CR), of which EPEC and EHEC are important causative agents of foodborne diseases worldwide. While A/E pathogen infections cause mild symptoms in the immunocompetent hosts, an increasing number of studies show that they produce more severe morbidity and mortality in immunocompromised and/or immunodeficient hosts. However, the pathogenic mechanisms and crucial host-pathogen interactions during A/E pathogen infections under immunocompromised conditions remain elusive. We performed a functional screening by infecting interleukin-22 (IL-22) knockout (Il22-/-) mice with a library of randomly mutated CR strains. Our screen reveals that interruption of the espF gene, which encodes the Type III Secretion System effector EspF (E. coli secreted protein F) conserved among A/E pathogens, completely abolishes the high mortality rates in CR-infected Il22-/- mice. Chromosomal deletion of espF in CR recapitulates the avirulent phenotype without impacting colonization and proliferation of CR, and EspF complement in ΔespF strain fully restores the virulence in mice. Moreover, the expression levels of the espF gene are elevated during CR infection and CR induces disruption of the tight junction (TJ) strands in colonic epithelium in an EspF-dependent manner. Distinct from EspF, chromosomal deletion of other known TJ-damaging effector genes espG and map failed to impede CR virulence in Il22-/- mice. Hence our findings unveil a critical pathophysiological function for EspF during CR infection in the immunocompromised host and provide new insights into the complex pathogenic mechanisms of A/E pathogens.


Asunto(s)
Proteínas Bacterianas/inmunología , Proteínas Portadoras/inmunología , Citrobacter rodentium/inmunología , Infecciones por Enterobacteriaceae/inmunología , Huésped Inmunocomprometido , Mucosa Intestinal/inmunología , Uniones Estrechas/inmunología , Animales , Proteínas Bacterianas/genética , Proteínas Portadoras/genética , Línea Celular , Citrobacter rodentium/genética , Citrobacter rodentium/patogenicidad , Colon/inmunología , Colon/microbiología , Colon/patología , Infecciones por Enterobacteriaceae/genética , Infecciones por Enterobacteriaceae/patología , Interleucinas/deficiencia , Interleucinas/inmunología , Mucosa Intestinal/microbiología , Mucosa Intestinal/patología , Ratones , Ratones Noqueados , Uniones Estrechas/genética , Uniones Estrechas/patología
14.
Fish Shellfish Immunol ; 92: 508-518, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31247319

RESUMEN

Mechanisms by which vaccines enhance immunity to combat bacterial pathogens are not fully understood. Recently, we have found that live Edwardsiella tarda vaccine enhances ability against the bacterial challenge by metabolic modulation in zebrafish. Here we first explored the metabolic modulation promoted by inactivated E. tarda to eliminate the pathogen. Inactivated E. tarda vaccine modulated a similar metabolome to combat with the pathogen in zebrafish as live E. tarda vaccine did. Specifically, both vaccines promoted biosynthesis of unsaturated fatty acids and the TCA cycle. However, due to relatively higher activated TCA cycle in inactivated vaccine than live vaccine, live vaccine promoted higher abundance of palmitate than inactivated vaccine. Consistently, the protection against E. tarda challenge was palmitate dose-dependent. Live vaccine activated higher expression of IL-1ß, IL-8,Cox-2 genes and lower expression of IL-15, IL-21 genes than inactivated vaccine, which is similar to the results stimulated by high and low doses of palmitate, respectively. These findings indicate live and inactivated E. tarda vaccines stimulate differential abundances of palmitate that contribute to differential innate immunities against bacterial infection. Thus, metabolic environment contributes to immune response.


Asunto(s)
Ácido Palmítico/metabolismo , Animales , Vacunas Bacterianas , Edwardsiella tarda/fisiología , Infecciones por Enterobacteriaceae/inmunología , Enfermedades de los Peces/inmunología , Distribución Aleatoria , Vacunas Atenuadas , Vacunas de Productos Inactivados , Pez Cebra
15.
Infect Immun ; 87(7)2019 07.
Artículo en Inglés | MEDLINE | ID: mdl-31061145

RESUMEN

Tissue-resident memory T cells (TRM cells) are a novel population of tissue-restricted antigen-specific T cells. TRM cells are induced by pathogens and promote host defense against secondary infections. Although TRM cells cannot be detected in circulation, they are the major memory CD4+ and CD8+ T-cell population in tissues in mice and humans. Murine models of CD8+ TRM cells have shown that CD8+ TRM cells maintain tissue residency via CD69 and though tumor growth factor ß-dependent induction of CD103. In contrast to CD8+ TRM cells, there are few models of CD4+ TRM cells. Thus, much less is known about the factors regulating the induction, maintenance, and host defense functions of CD4+ TRM cells. Citrobacter rodentium is known to induce IL-17+ and IL-22+ CD4+ T cells (Th17 and Th22 cells, respectively). Moreover, data from IL-22 reporter mice show that most IL-22+ cells in the colon 3 months after C. rodentium infection are CD4+ T cells. This collectively suggests that C. rodentium may induce CD4+ TRM cells. Here, we demonstrate that C. rodentium induces a population of IL-17A+ CD4+ T cells that are tissue restricted and antigen specific, thus meeting the criteria of CD4+ TRM cells. These cells expand and are a major source of IL-22 during secondary C. rodentium infection, even before the T-cell phase of the host response in primary infection. Finally, using FTY 720, which depletes circulating naive and effector T cells but not tissue-restricted T cells, we show that these CD4+ TRM cells can promote host defense.


Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Citrobacter rodentium/inmunología , Infecciones por Enterobacteriaceae/inmunología , Animales , Citrobacter rodentium/genética , Infecciones por Enterobacteriaceae/genética , Infecciones por Enterobacteriaceae/microbiología , Humanos , Memoria Inmunológica , Interleucina-17/genética , Interleucina-17/inmunología , Interleucinas/genética , Interleucinas/inmunología , Ratones , Ratones Endogámicos C57BL , Células Th17/inmunología
16.
Mol Immunol ; 112: 10-21, 2019 08.
Artículo en Inglés | MEDLINE | ID: mdl-31075558

RESUMEN

Extracellular adenosine triphosphate (eATP), released following inflammatory stimulation or infection, is a potent signaling molecule in activating innate immune responses in fish. However, the regulation of eATP-mediated innate immunity in fish remains unknown. Ecto-nucleoside triphosphate diphosphohydrolase 1 (CD39) is a critical molecular switch for controlling the ATP levels in the extracellular space. CD39 plays a key role in regulating eATP-activated innate immune responses through the phosphohydrolysis of pro-inflammatory eATP to inactive AMP. Here, we identified and characterized a CD39 homolog (namely, poCD39) in the Japanese flounder Paralichthys olivaceus and analyzed its regulatory role in eATP-mediated innate immunity. Real-time quantitative PCR analysis revealed that poCD39 is ubiquitously present in all tested normal tissues with dominant expression in enriched Japanese flounder head kidney macrophages (HKMs). Immune challenge experiments demonstrated that poCD39 expression was upregulated by inflammatory stimulation and Edwardsiella tarda infection. Biochemical and immunofluorescence analysis revealed that poCD39 is a functional glycosylated membrane protein for the hydrolysis of eATP. Inhibition of poCD939 activity with the ecto-NTPDase inhibitor ARL 67156 resulted in increased IL-1beta gene expression and ROS production in Japanese flounder HKMs. In contrast, overexpression of poCD39 in Japanese flounder FG-9307 cells reduced eATP-induced pro-inflammatory cytokine IL-1beta gene expression. Finally, poCD39 expression was significantly induced by eATP stimulation in the HKMs, suggesting that eATP may provide a feedback mechanism for transcriptional regulation of fish CD39. Taken together, we identified and characterized a functional fish CD39 protein involved in regulating eATP-mediated innate immune responses in fish.


Asunto(s)
Adenosina Trifosfato/inmunología , Antígenos CD/inmunología , Apirasa/inmunología , Lenguado/inmunología , Inmunidad Innata/inmunología , Adenosina Monofosfato/inmunología , Animales , Edwardsiella tarda/inmunología , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/microbiología , Infecciones por Enterobacteriaceae/veterinaria , Proteínas de Peces/inmunología , Lenguado/microbiología , Regulación de la Expresión Génica/inmunología , Riñón Cefálico/inmunología , Riñón Cefálico/microbiología , Inflamación/inmunología , Inflamación/microbiología , Japón , Macrófagos/inmunología , Macrófagos/microbiología , Especies Reactivas de Oxígeno/inmunología , Transcripción Genética/inmunología , Regulación hacia Arriba/inmunología
17.
Fish Shellfish Immunol ; 92: 151-164, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31108176

RESUMEN

IL-12 is an important cytokine that connects the innate and adaptive immune systems. The complete gene structure of olive flounder IL-12 and its characteristics have not yet been formally reported. Here, we report the complete sequences of both subunits of olive flounder IL-12 (IL-12p35 and IL-12p40). In addition, its function was analyzed by generating the single-chain rIL-12 of which subunits were fused by a GS linker and the rIL-12-specific mouse antibody. The cDNA sequences of IL-12p35 and IL-12p40 were 1059 nucleotides and 1319 nucleotides, respectively. The analyses of their gene structures, deduced amino acid sequences, protein model structures, and phylogenetic trees confirmed the accurate identification of olive flounder IL-12. The protein structure model suggested that an inter-subunit disulfide bond might be formed between the Cys177 of p35 and Cys74 of p40 to link the subunits. Olive flounder expressed IL-12p40 at higher levels than IL-12p35 in the various tissues under natural conditions although both expression levels were low. However, when infected by Edwardsiella tarda or stimulated by LPS, the flounder expressed both of the subunit genes at similar maximized levels in 6 h and gradually reduced thereafter. Olive flounder PBMC induced with the rIL-12 increased IFN-γ and TNF-α expression but decreased IL-10 expression as did treatment with LPS. However, when the LPS-treated PBMC were neutralized with the rIL-12-specific antibody, the pattern of cytokine expression was precisely reversed. In conclusion, we have formally identified the gene structure and function of olive flounder IL-12.


Asunto(s)
Inmunidad Adaptativa/genética , Enfermedades de los Peces/inmunología , Peces Planos/genética , Peces Planos/inmunología , Inmunidad Innata/genética , Interleucina-12/genética , Interleucina-12/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Edwardsiella tarda/fisiología , Infecciones por Enterobacteriaceae/inmunología , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Lenguado/genética , Lenguado/inmunología , Perfilación de la Expresión Génica/veterinaria , Interleucina-12/química , Lipopolisacáridos/farmacología , Filogenia , Alineación de Secuencia/veterinaria
18.
Fish Shellfish Immunol ; 92: 851-860, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31129187

RESUMEN

Innate immunity is regulated by phagocytic cells and is critical for host control of bacterial infection. In many bacteria, the type VI secretion system (T6SS) can affect bacterial virulence in certain environments, but little is known about the mechanisms underlying T6SS regulation of innate immune responses during infection in vivo. Here, we developed an infection model by microinjecting bacteria into the tail vein muscle of 3-day-post-fertilized zebrafish larvae, and found that both macrophages and neutrophils are essential for bacterial clearance. Further study revealed that EvpP plays a critical role in promoting the pathogenesis of Edwardsiella piscicida (E. piscicida) via inhibiting the phosphorylation of Jnk signaling to reduce the expression of chemokine (CXC motif) ligand 8 (cxcl8a), matrix metallopeptidase 13 (mmp13) and interleukin-1ß (IL-1ß) in vivo. Subsequently, by utilizing Tg (mpo:eGFP+/+) zebrafish larvae for E. piscicida infection, we found that the EvpP-inhibited Jnk-caspy (caspase-1 homolog) inflammasome signaling axis significantly suppressed the recruitment of neutrophils to infection sites, and the caspy- or IL-1ß-morpholino (MO) knockdown larvae were more susceptible to infection and failed to restrict bacterial colonization in vivo. taken together, this interaction improves our understanding about the complex and contextual role of a bacterial T6SS effector in modulating the action of neutrophils during infection, and offers new insights into the warfare between bacterial weapons and host immunological surveillance.


Asunto(s)
Proteínas Bacterianas/fisiología , Inmunidad Innata , Inflamasomas/inmunología , Macrófagos/metabolismo , Neutrófilos/metabolismo , Pez Cebra/inmunología , Animales , Edwardsiella/fisiología , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/veterinaria , Enfermedades de los Peces/inmunología , Sistemas de Secreción Tipo VI/fisiología
19.
Microb Pathog ; 131: 164-169, 2019 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-30978428

RESUMEN

Bacterial diseases are one of the major problems in freshwater fish culture and have been linked to significant losses and high mortality rate. In this study, Nile tilapia Oreochromis niloticus was infected by Providencia rettgeri to evaluate the oxidative stress and antioxidant responses in the fish tissues. Juvenile Nile tilapia was divided into two groups, as follow: control (uninfected) and experimentally infected with 100 µL of P. rettgeri suspension containing 2.4 × 107 viable cells/fish, and the liver and kidney tissues were collected on days 7 and 14 post-infection (PI). Liver and kidney ROS and lipid peroxidation levels were high in infected fish on day 14 PI compared to control group, while superoxide dismutase activity was lower in liver (days 7 and 14 PI) and kidney (day 14 PI) compared to their respective control groups. Liver and kidney antioxidant capacity against peroxyl radicals, non-proteic, and proteic thiols levels was lower in infected tilapia on day 14 PI compared to control group. Based on these results, P. rettgeri infection may elicit oxidative damage via increased ROS production, decreased ROS elimination and inhibits enzymatic and non-enzymatic antioxidant defense systems; which may contribute directly to disease pathophysiology of infected animals.


Asunto(s)
Antioxidantes/metabolismo , Cíclidos/metabolismo , Enfermedades de los Peces/metabolismo , Estrés Oxidativo , Providencia/patogenicidad , Animales , Brasil , Cíclidos/inmunología , Cíclidos/microbiología , Modelos Animales de Enfermedad , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/microbiología , Infecciones por Enterobacteriaceae/veterinaria , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/microbiología , Interacciones Huésped-Patógeno/fisiología , Riñón/metabolismo , Peroxidación de Lípido , Hígado/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Tilapia/microbiología
20.
PLoS Pathog ; 15(4): e1007719, 2019 04.
Artículo en Inglés | MEDLINE | ID: mdl-30973939

RESUMEN

The regulation of mucosal immune function is critical to host protection from enteric pathogens but is incompletely understood. The nervous system and the neurotransmitter acetylcholine play an integral part in host defense against enteric bacterial pathogens. Here we report that acetylcholine producing-T-cells, as a non-neuronal source of ACh, were recruited to the colon during infection with the mouse pathogen Citrobacter rodentium. These ChAT+ T-cells did not exclusively belong to one Th subset and were able to produce IFNγ, IL-17A and IL-22. To interrogate the possible protective effect of acetylcholine released from these cells during enteric infection, T-cells were rendered deficient in their ability to produce acetylcholine through a conditional gene knockout approach. Significantly increased C. rodentium burden was observed in the colon from conditional KO (cKO) compared to WT mice at 10 days post-infection. This increased bacterial burden in cKO mice was associated with increased expression of the cytokines IL-1ß, IL-6, and TNFα, but without significant changes in T-cell and ILC associated IL-17A, IL-22, and IFNγ, or epithelial expression of antimicrobial peptides, compared to WT mice. Despite the increased expression of pro-inflammatory cytokines during C. rodentium infection, inducible nitric oxide synthase (Nos2) expression was significantly reduced in intestinal epithelial cells of ChAT T-cell cKO mice 10 days post-infection. Additionally, a cholinergic agonist enhanced IFNγ-induced Nos2 expression in intestinal epithelial cell in vitro. These findings demonstrated that acetylcholine, produced by specialized T-cells that are recruited during C. rodentium infection, are a key mediator in host-microbe interactions and mucosal defenses.


Asunto(s)
Acetilcolina/metabolismo , Citrobacter rodentium/inmunología , Colon/inmunología , Infecciones por Enterobacteriaceae/inmunología , Linfocitos T/inmunología , Animales , Células Cultivadas , Colon/metabolismo , Citocinas/metabolismo , Infecciones por Enterobacteriaceae/microbiología , Interleucina-17/metabolismo , Mucosa Intestinal/inmunología , Mucosa Intestinal/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores CXCR5/fisiología
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