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2.
Comp Biochem Physiol B Biochem Mol Biol ; 254: 110575, 2021 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-33609806

RESUMO

Hypoxia-inducible factor-1α (HIF-1α) plays a critical role in immune and inflammatory responses and is important in controlling a variety of processes in monocytes and macrophages. However, the role of HIF-1α in the teleost immune system remains less known. In this study, we cloned the cDNA sequence of HIF-1α from the ayu (Plecoglossus altivelis, PaHIF-1α). Sequence and phylogenetic tree analysis showed that PaHIF-1α clustered within the fish HIF-1α tree and was closely related to that of Northern pike (Esox lucius). PaHIF-1α was expressed in all tested tissues and expression increased in liver, head kidney, and body kidney upon Vibrio anguillarum infection. PaHIF-1α was found to regulate the expression of cytokines in ayu monocytes/macrophages (MO/MФ). PaHIF-1α mediated hypoxia-induced enhancement of MO/MФ phagocytic and bactericidal activities to enhance host defenses. Compared with the control, intermittent hypoxia further increased the expression of PaHIF-1α mRNA, improved the survival rate, and reduced the bacterial load of V. anguillarum-infected ayu. Therefore, PaHIF-1α may play a predominant role in the modulation of ayu MO/MФ function.

3.
Mol Immunol ; 133: 1-13, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33610121

RESUMO

Transcription factor PU.1 is a regulator of macrophage function, however, the specific function of PU.1 in teleost monocytes/macrophages (MO/MФ) remains unknown. We determined the cDNA sequence of two PU.1 genes from ayu (Plecoglossus altivelis; PaPU.1a and PaPU.1b). Sequence comparisons showed that PaPU.1 were most closely related to the PU.1 of rainbow smelt (Osmerus mordax). The PU.1 transcripts were mainly expressed in the spleen, and their expression was altered in various tissues upon infection with Vibrio anguillarum. PaPU.1a and PaPU.1b proteins were upregulated in MO/MФ, after infection. RNA interference was employed to knockdown PaPU.1a and PaPU.1b to investigate their function in MO/MФ. The expression of inflammatory cytokines was regulated by PaPU.1a, but not PaPU.1b, in ayu MO/MФ upon V. anguillarum infection. Both PaPU.1a and PaPU.1b knockdown lowered the phagocytic activity of MO/MФ. Furthermore, PaPU.1b knockdown attenuated MO/MФ bacterial killing capability. Our results indicate that two PaPU.1 genes differentially modulate the immune response in ayu MO/MФ against bacterial infection.

4.
J Hematol Oncol ; 14(1): 30, 2021 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-33596982

RESUMO

Histone methylation is a key posttranslational modification of chromatin, and its dysregulation affects a wide array of nuclear activities including the maintenance of genome integrity, transcriptional regulation, and epigenetic inheritance. Variations in the pattern of histone methylation influence both physiological and pathological events. Lysine-specific demethylase 5A (KDM5A, also known as JARID1A or RBP2) is a KDM5 Jumonji histone demethylase subfamily member that erases di- and tri-methyl groups from lysine 4 of histone H3. Emerging studies indicate that KDM5A is responsible for driving multiple human diseases, particularly cancers. In this review, we summarize the roles of KDM5A in human cancers, survey the field of KDM5A inhibitors including their anticancer activity and modes of action, and the current challenges and potential opportunities of this field.

5.
Dev Comp Immunol ; 117: 103978, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33338518

RESUMO

Urocortin (UCN) is a hormone in the hypothalamic-pituitary-adrenal axis that is expressed in various immune cells. However, the function of teleost UCN in the immune system remains unclear. In this study, we cloned the cDNA sequence of UCN from ayu Plecoglossus altivelis (PaUCN). Sequence and phylogenetic tree analyses showed that PaUCN clustered within the fish UCN 1 group and was most related to the rainbow trout (Oncorhynchus mykiss) UCN. PaUCN was expressed in all tested tissues and its expression increased in the liver, spleen, head kidney, and gill upon Vibrio anguillarum infection. Mature PaUCN protein (mPaUCN) treatment affected the phagocytosis and bacterial killing of monocytes/macrophages (MO/MФ). mPaUCN reduced pro-inflammatory cytokine expression in MO/MФ, which was partially mediated via interaction with ayu interleukin-6. mPaUCN reduced bacterial load and increased the survival of V. anguillarum-infected ayu. Overall, UCN as an endocrine factor regulates the immune response of ayu after infection by activating MO/MФ, thus contributing to enhance fish survival.

6.
7.
Dev Comp Immunol ; : 103930, 2020 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-33212093

RESUMO

Virus-encoded tumor necrosis factor receptors (vTNFRs) facilitate viral escape from the host immune response during viral propagation. Cyprinid Herpesvirus-2 (CyHV-2) is a double-stranded DNA virus of alloherpesviridae family that causes great economic losses in the aquaculture industry. The present study identified and characterized a novel TNFR homolog termed ORF4 in CyHV-2. ORF4 was identified as a secreted protein and a homolog of herpesvirus entry mediator (HVEM). ORF4 localized to the cytoplasm in infected GiCF cells. ORF4 overexpression enhanced viral propagation, while downregulation of ORF4 via siRNA decreased viral propagation. ORF4 overexpression promoted GiCF proliferation, and its downregulation suppressed CyHV-2-induced apoptosis. GST-pulldown and LC-MS/MS assays identified 44 conditional binding proteins that interact with ORF4 protein, while the GST pulldown test did not support the idea that ORF4 interact with histone H3.3. Taken together, our results contribute to our understanding of the vTNFR function in alloherpesviridae pathogenesis and host immune regulation.

8.
J Immunol ; 204(5): 1225-1241, 2020 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-31959734

RESUMO

Hematopoietic stem/progenitor cells (HSPCs) generate the entire repertoire of immune cells in vertebrates and play a crucial role during infection. Although two copies of CXC motif chemokine receptor 4 (CXCR4) genes are generally identified in teleosts, the function of teleost CXCR4 genes in HSPCs is less known. In this study, we identified two CXCR4 genes from a teleost, ayu (Plecoglossus altivelis), named PaCXCR4a and PaCXCR4b. PaCXCR4b was constitutively expressed in ayu HSPCs, whereas PaCXCR4a was induced by LPS treatment. The stromal-derived factor-1-binding activity of CXCR4b was significantly higher than that of CXCR4a, whereas the LPS-binding activity of CXCR4a was significantly higher than that of CXCR4b in the teleosts ayu, large yellow croaker (Larimichthys crocea), and tiger puffer (Takifugu rubripes). CXCR4a+ HSPCs were mobilized into blood by LPS, whereas CXCR4b+ HSPCs were mobilized by leukocyte cell-derived chemotaxin-2. PaSDF-1 and PaCXCR4b, but not PaCXCR4a, inhibited HSPC proliferation by regulating reactive oxygen species levels. Compared with PaCXCR4b+ HSPCs, PaCXCR4a+ HSPCs preferentially differentiated into myeloid cells in ayu by maintaining high stem cell leukemia expression. These data suggest that the two copies of CXCR4s achieve a division of labor in the regulation of teleost HSPC homeostasis, supporting the concept that subfunctionalization after gene duplication in teleosts may stabilize the immune system.


Assuntos
Proliferação de Células/fisiologia , Proteínas de Peixes/imunologia , Células-Tronco Hematopoéticas/imunologia , Homeostase/imunologia , Perciformes/imunologia , Receptores CXCR4/imunologia , Animais , Proteínas de Peixes/genética , Células-Tronco Hematopoéticas/citologia , Homeostase/genética , Leucócitos/citologia , Leucócitos/imunologia , Perciformes/genética , Espécies Reativas de Oxigênio/imunologia , Receptores CXCR4/genética , Especificidade da Espécie , Tetraodontiformes/genética , Tetraodontiformes/imunologia
9.
Dev Comp Immunol ; 103: 103513, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31585194

RESUMO

Adrenergic receptors (ARs) are members of the G-protein-coupled receptor superfamily that can be categorized into αARs and ßARs. The specific function of ARs in teleost monocytes/macrophages (MO/MФ) remains unknown. We determined the cDNA sequence of ARs from ayu (Plecoglossus altivelis; PaαAR and PaßAR). Sequence comparisons showed that PaαAR was most closely related to the αAR of the Japanese flounder and Nile tilapia, while PaßAR was most closely related to the ßAR of Atlantic salmon. The AR transcripts were mainly expressed in the spleen, and their expression was altered in various tissues upon infection with Vibrio anguillarum. PaαAR and PaßAR proteins were upregulated in MO/MФ after infection, and PaßAR knockdown resulted in a pro-inflammatory status in ayu MO/MФ upon V. anguillarum infection and lowered the phagocytic activity of MO/MФ. Our results indicate that PaßAR plays the role of an anti-inflammatory mediator in the immune response of ayu against bacterial infection.


Assuntos
Proteínas de Peixes/imunologia , Osmeriformes/imunologia , Receptores Adrenérgicos beta/imunologia , Animais , Doenças dos Peixes/imunologia , Macrófagos/imunologia , Monócitos/imunologia , Vibrio , Vibrioses/imunologia , Vibrioses/veterinária
10.
Fish Shellfish Immunol ; 93: 191-199, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31326589

RESUMO

Interleukin-6 (IL-6) is one of the most pleiotropic cytokines because of its wide range of effects on cells of the immune and non-immune systems in the body. However, the role of IL-6 in fish monocytes/macrophages (MO/MФ) is poorly understood. In this study, we cloned the cDNA sequence of the IL-6 gene from ayu (Plecoglossus altivelis) and demonstrated using a tissue distribution assay that ayu interleukin-6 (PaIL-6) mRNA is expressed in all tested tissues. Changes in expression were observed in immune tissues as well as in MO/MФ after a Vibrio anguillarum infection; subsequently, PaIL-6 was expressed and purified to prepare anti-PaIL-6 antibodies. Recombinant PaIL-6 protein (rPaIL-6) treatment enhanced pro-inflammatory cytokine expression. Ayu interleukin-6 receptor ß (PaIL-6Rß) knockdown resulted in decreased pro-inflammatory cytokine expression in MO/MФ treated with rPaIL-6, whereas no significant changes were observed after ayu interleukin-6 receptor α (PaIL-6Rα) knockdown in MO/MФ. PaIL-6 and PaIL-6Rß knockdown in MO/MФ inhibited the phosphorylation of signal transducer and activator of transcription 1. Moreover, PaIL-6Rß knockdown inhibited the phagocytic and bactericidal ability of ayu MO/MФ treated with rPaIL-6. These data indicate that PaIL-6 may be able to regulate the function of ayu MO/MФ.


Assuntos
Receptor gp130 de Citocina/genética , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Interleucina-6/genética , Osmeriformes/genética , Osmeriformes/imunologia , Sequência de Aminoácidos , Animais , Receptor gp130 de Citocina/imunologia , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Técnicas de Silenciamento de Genes/veterinária , Interleucina-6/química , Interleucina-6/imunologia , Macrófagos/imunologia , Monócitos/imunologia , Fagocitose/genética , Fagocitose/imunologia , Fosforilação , Filogenia , Fator de Transcrição STAT1/metabolismo , Alinhamento de Sequência/veterinária , Vibrio/fisiologia , Vibrioses/imunologia , Vibrioses/veterinária
11.
Zool Res ; 40(3): 146-150, 2019 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-31011129

RESUMO

Macrophages exist in most tissues and play a variety of functions in vertebrates. Teleost fish species are found in most aquatic environments throughout the world and are quite diverse for a group of vertebrate animals. Due to whole genome duplication and environmental adaptation, teleost monocytes/macrophages possess a variety of different functions and modulations compared with those of mammals. A deeper understanding of teleost monocytes/macrophages in the immune system will not only help develop teleost-specific methods of disease prevention but will also help improve our understanding of the various immune mechanisms in mammals. In this review, we summarize the differences in polarization and phagocytosis of teleost and mammalian macrophages to improve our understanding of the various immune mechanisms in vertebrates.


Assuntos
Peixes , Macrófagos/fisiologia , Monócitos/fisiologia , Fagocitose/fisiologia , Animais
12.
Fish Shellfish Immunol ; 86: 1151-1161, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30597252

RESUMO

The glucocorticoid receptor (GR) is an important feedback regulator of the hypothalamic-pituitary-interrenal (HPI) axis. However, there are a limited number of studies focused on host-pathogen interactions in which an association between GR and immune response has been evaluated in monocytes/macrophages (MO/MФ) after being challenged with highly pathogenic bacteria. Here, we cloned the cDNA sequence of the glucocorticoid receptor (PaGR) gene from ayu fish. The PaGR transcript was expressed in all tissues, and changes in expression were observed in immune tissues and MO/MФ after live Vibrio anguillarum infection. Subsequently, PaGR was expressed and purified to prepare anti-PaGR antibodies. We analyzed the subcellular localization of PaGR. PaGR was expressed not only in the intracellular space but also in the plasma membrane. PaGR activation decreased the expression of pro-inflammatory cytokines and increased the expression of anti-inflammatory cytokines. However, PaGR activation suppressed the phagocytosis activity of V. anguillarum-infected ayu MO/MФ via a non-genomic pathway. Interestingly, PaGR activation could enhance MO/MФ bacterial killing capability and apoptosis. Therefore, PaGR may modulate the immune response in ayu MO/MФ by genomic and non-genomic pathways.


Assuntos
Macrófagos/imunologia , Monócitos/imunologia , Receptores de Glucocorticoides/genética , Receptores de Glucocorticoides/metabolismo , Vibrioses/veterinária , Animais , Apoptose/imunologia , Membrana Celular/metabolismo , Doenças dos Peixes/imunologia , Osmeriformes , Fagocitose/imunologia , Vibrio , Vibrioses/genética , Vibrioses/imunologia
13.
Front Immunol ; 9: 2758, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30542348

RESUMO

Macrophages in teleosts are less sensitive to lipopolysaccharide (LPS) compared to mammals. The functional equivalent of the mammalian LPS surface receptor in teleost macrophages for the pro-inflammatory response is either non-existent or replaced by negative regulation. LPS signaling in teleost macrophages remains unclear. Here, we found a scavenger receptor class B 2a (PaSRB2a) that played a crucial role in LPS signaling in teleost macrophages. The internalization of LPS and subsequent pro-inflammatory responses in macrophages were mediated by PaSRB2a, which is a novel isoform of the mammalian SRB2 gene. LPS internalization by PaSRB2a is dependent on its C-terminal intracellular domain. Following LPS internalization, it interacts with the ayu intracellular receptors nucleotide-binding oligomerization domain protein 1 (PaNOD1) and PaNOD2. Moreover, LPS pre-stimulation with sub-threshold concentrations reduced the effect of secondary LPS treatment on pro-inflammatory responses that were mediated by PaSRB2a. The pro-inflammatory responses in LPS-treated ayu were down-regulated upon PaSRB2a knockdown by lentivirus siRNA delivery. In grass carp and spotted green pufferfish, SRB2a also mediated LPS internalization and pro-inflammatory responses. Our work identifies a novel LPS signaling pathway in teleosts that differs from those in mammals, and contributes to our understanding of the evolution of pathogen recognition in vertebrates.


Assuntos
Lipopolissacarídeos/metabolismo , Macrófagos/metabolismo , Animais , Carpas/metabolismo , Regulação para Baixo/fisiologia , Inflamação/metabolismo , Receptores de Lipopolissacarídeos/metabolismo , Osmeriformes/metabolismo , Transdução de Sinais/fisiologia , Tetraodontiformes/metabolismo
14.
Fish Shellfish Immunol ; 82: 319-329, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30130657

RESUMO

Adiponectin (ADP), a regulator of the innate immune system, plays a role in the progression of inflammation and metabolic disorders in mammals. However, the role of ADP in fish is poorly understood. Here, we cloned the cDNA sequence of a ADP homolog (PaADP) gene from ayu. Multiple sequence alignment revealed that PaADP presented typical characteristics of ADPs. Phylogenetic tree analysis showed that PaADP was most closely related to that of rainbow trout. In healthy ayu, the transcripts of PaADP were detected in most of the tested tissues and cells, with the highest level in the adipose tissue. Upon V. anguillarum infection, the mRNA expression of PaADP was significantly up-regulated in the tissues and cells except adipose tissue. Subsequently, the full-length mature PaADP (fPaADP) and the globular domain fragment (gPaADP) were prokaryotically expressed in bacteria and purified, and anti-PaADP antibodies were produced. Western blot analysis revealed that three fragments including fPaADP and gPaADP were existed in ayu serum. The recombinant fPaADP (rfPaADP) had an anti-inflammatory effect on ayu MO/MФ by upregulating anti-inflammatory cytokine expressions, downregulating pro-inflammatory cytokine expressions, inhibiting the phagocytosis and subsequent bacterial killing. In contrast, the recombinant gPaADP (rgPaADP) presented a pro-inflammatory effect on ayu MO/MФ by upregulating pro-inflammatory cytokine expression, downregulating anti-inflammatory cytokine expressions, enhancing the phagocytosis and subsequent bacterial killing. These results suggested that fPaADP and gPaADP have opposite roles in the regulation of MO/MФ functions in ayu.


Assuntos
Adiponectina/genética , Adiponectina/imunologia , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Osmeriformes/genética , Osmeriformes/imunologia , Adiponectina/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Filogenia , Alinhamento de Sequência/veterinária , Vibrio/fisiologia , Vibrioses/imunologia
15.
Dev Comp Immunol ; 84: 37-47, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29408399

RESUMO

Tissue factor (TF) plays an important role in the host's immune system as the principal initiator of coagulation. However, the precise function of TF in teleosts remains unclear. We determined the cDNA sequence of TF from ayu Plecoglossus altivelis (PaTF). The PaTF transcript was expressed in all tested tissues, and changes in expression were observed in tissues and monocytes/macrophages (MO/MФ) upon infection with Vibrio anguillarum. PaTF was prokaryotically expressed and purified to prepare anti-PaTF antibodies. Western blot analysis revealed that native PaTF was glycosylated in thrombocytes, but not in ayu MO/MФ. Microparticles could transfer PaTF to thrombocytes. PaTF neutralization or knockdown led to anti-inflammatory status in ayu MO/MФ upon V. anguillarum infection. PaTF neutralization reduced the apoptosis of ayu MO/MФ and improve survival rate in V. anguillarum-infected ayu. Our results indicate that PaTF plays a role in ayu immune response against bacterial infection as a pro-inflammatory mediator.


Assuntos
Proteínas de Peixes/genética , Macrófagos/imunologia , Monócitos/imunologia , Osmeriformes/imunologia , Tromboplastina/genética , Vibrioses/imunologia , Vibrio alginolyticus/fisiologia , Animais , Apoptose , Células Cultivadas , Clonagem Molecular , Proteínas de Peixes/metabolismo , Técnicas de Silenciamento de Genes , Imunidade Inata , Imunomodulação , Mediadores da Inflamação/metabolismo , Tromboplastina/metabolismo
16.
Fish Shellfish Immunol ; 72: 301-308, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29128493

RESUMO

The CC motif chemokine 19 (CCL19) functions in acute inflammation by recruiting lymphocytes and other cells. However, CCL19 has only been investigated in few fish species. In this study, we characterized a CCL19-like molecule (PaCCL19l) in ayu (Plecoglossus altivelis), a teleost fish. Sequence analysis revealed that PaCCL19l was most closely related to Atlantic salmon (Salmon salar) CCL19l1, which belonged to the fish CCL19a.1 subcluster. PaCCL19l was constitutively expressed in the tested ayu tissues and peripheral blood mononuclear cells (PBMCs), with the highest transcript level in PBMCs. Upon infection with Vibrio anguillarum, the expressions of PaCCL19l in the head kidney, liver, spleen, PBMCs, and monocytes/macrophages (MO/MΦ) were dramatically up-regulated. Recombinant PaCCL19l (rPaCCL19l) exhibited a significant effect on the chemotaxis of lymphocytes and MO/MΦ in vitro and in vivo. Meanwhile, rPaCCL19l exerted a high chemotaxic activity for lipopolysaccharide (LPS)-stimulated MO/MΦ (M1-type), but not for cyclic adenosine monophosphate (cAMP)-stimulated MO/MΦ (M2-type). When ayu MO/MΦ was treated with rPaCCL19l along with Vibrio anguillarum infection, the mRNA expression of proinflammatory cytokines (IL-1ß, TNFα, IL-6, IL-12b, and IFN-γ) was up-regulated, while that of anti-inflammatory cytokines (IL-10, TGFß, and IL-22) was down-regulated. Ayu MO/MΦ treated with anti-PaCCL19l IgG gave the opposite result. These results implicated that PaCCL19l is involved in the selective chemotaxis of ayu immune cells and promotes the host at a pro-inflammatory state.


Assuntos
Quimiocina CCL19/genética , Quimiocina CCL19/imunologia , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Osmeriformes/genética , Osmeriformes/imunologia , Sequência de Aminoácidos , Animais , Quimiocina CCL19/química , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica , Leucócitos/metabolismo , Filogenia , Alinhamento de Sequência/veterinária , Vibrio/fisiologia , Vibrioses/imunologia
17.
J Immunol ; 198(12): 4692-4706, 2017 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-28500070

RESUMO

The study of multiple copies of chemokine receptor genes in various teleosts has long appealed to investigators seeking to understand the evolution of the immune system. The CXCR CXCR3 gene has two isoforms, CXCR3.1 and CXCR3.2, which are both expressed in macrophages. The distinct roles of teleost CXCR3s have not been identified previously. In this article, we found that CXCR3.1 and CXCR3.2 differentially contributed to macrophage polarization in the teleosts: ayu (Plecoglossus altivelis), grass carp (Ctenopharyngodon idella), and spotted green pufferfish (Tetraodon nigroviridis). In ayu macrophages, the P. altivelis CXCR3.1 (PaCXCR3.1) gene was constitutively expressed, whereas the P. altivelis CXCR3.2 (PaCXCR3.2) gene was induced postinfection with Escherichia coli Upon E. coli infection, PaCXCR3.1+ and PaCXCR3.2+ macrophages showed an M1 and an M2 phenotype, respectively. CXCL9-11-like proteins mediated M1 and M2 polarization by interacting with the PaCXCR3.1 and PaCXCR3.2 proteins on macrophages, respectively. The transcription factors P. altivelis STAT1 and P. altivelis STAT3 were activated in PaCXCR3.1+ and PaCXCR3.2+ macrophages, respectively. Furthermore, the prognosis of septic ayu adoptively transferred with PaCXCR3.2+ macrophages was improved. Our data reveal a previously unknown mechanism for macrophage polarization, suggesting that redundant genes may regulate crucial functions in the teleost immune system.


Assuntos
Carpas/imunologia , Proteínas de Peixes/genética , Macrófagos/fisiologia , Osmeriformes/imunologia , Receptores CXCR3/genética , Tetraodontiformes/imunologia , Animais , Carpas/genética , Carpas/metabolismo , Diferenciação Celular , Clonagem Molecular , Proteínas de Peixes/metabolismo , Peixes/classificação , Peixes/imunologia , Regulação da Expressão Gênica , Macrófagos/imunologia , Monócitos/imunologia , Osmeriformes/genética , Osmeriformes/metabolismo , Fagocitose , Filogenia , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Receptores CXCR3/metabolismo , Fator de Transcrição STAT1/genética , Fator de Transcrição STAT1/metabolismo , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Tetraodontiformes/genética , Tetraodontiformes/metabolismo
18.
PLoS One ; 12(5): e0177960, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28542591

RESUMO

Hypoxia-inducible factor-1α (HIF-1α) plays a critical role in immune and inflammatory responses and is important in controlling a variety of processes in monocytes and macrophages. However, very little information is available about the functions of HIF-1α in fish monocytes/macrophages (MO/MФ). In this study, the cDNA sequence of the mudskipper (Boleophthalmus pectinirostris) HIF-1α gene (BpHIF-1α) was determined. Sequence comparison and phylogenetic tree analysis showed that BpHIF-1α is clustered in the fish HIF-1α tree. Constitutive expression of BpHIF-1α mRNA was detected by real-time quantitative PCR in all tested tissues, and the expression was found to be dramatically increased in the skin, liver, spleen, and kidney after Edwardsiella tarda infection. In addition, hypoxia and infection induced the expression of the BpHIF-1α transcript and protein in MO/MФ, respectively. Hypoxia caused an increase in phagocytic and bactericidal capacity of mudskipper MO/MФ in a BpHIF-1α-dependent manner. BpHIF-1α induced an anti-inflammatory status in MO/MФ upon E. tarda infection and hypoxia. Therefore, BpHIF-1α may play a predominant role in the modulation of mudskipper MO/MФ function in the innate immune system.


Assuntos
Edwardsiella tarda/imunologia , Infecções por Enterobacteriaceae/imunologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Macrófagos/imunologia , Monócitos/imunologia , Fagocitose/imunologia , Sequência de Aminoácidos/genética , Animais , Sequência de Bases , Hipóxia Celular/fisiologia , Células Cultivadas , Clonagem Molecular , Infecções por Enterobacteriaceae/microbiologia , Peixes , Imunidade Inata/imunologia , Inflamação/imunologia , Fígado/metabolismo , Interferência de RNA , RNA Interferente Pequeno/genética , Alinhamento de Sequência , Análise de Sequência de DNA , Pele/metabolismo
19.
Fish Shellfish Immunol ; 62: 257-264, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28130078

RESUMO

Monocytes/macrophages (MO/MФ) play an important role in the response to infection in Plecoglossus altivelis (ayu). However, the role of transcription factors in the function of ayu MO/MФ is poorly understood. Here, we cloned the cDNA sequence of the Kruppel-like factor 2 (PaKLF2) gene from ayu. Phylogenetic analysis indicated that PaKLF2 was closest to that of Atlantic salmon (Salmo salar). Real time quantitative PCR (RT-qPCR) revealed that the PaKLF2 mRNA level was highest in the peripheral blood mononuclear cells among all tested tissues. The mRNA expression of PaKLF2 was upregulated in the head kidney, liver, spleen, and brain after Listonella anguillarum infection. Subsequently, PaKLF2 was expressed and purified to prepare anti-PaKLF2 antibodies. After L. anguillarum challenge, the PaKLF2 mRNA and protein levels were significantly upregulated in ayu MO/MФ. Moreover, PaKLF2 knockdown in MO/MФ resulted in the enhancement of cytokine production as well as phagocytotic and bactericidal capability. Therefore, PaKLF2 may modulate the immune response in ayu by suppressing the function of MO/MФ.


Assuntos
Doenças dos Peixes/genética , Proteínas de Peixes/genética , Fatores de Transcrição Kruppel-Like/genética , Ativação de Macrófagos , Osmeriformes , Vibrioses/veterinária , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/química , Proteínas de Peixes/metabolismo , Fatores de Transcrição Kruppel-Like/química , Fatores de Transcrição Kruppel-Like/metabolismo , Macrófagos/imunologia , Monócitos/imunologia , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Alinhamento de Sequência/veterinária , Regulação para Cima , Vibrio/fisiologia , Vibrioses/genética , Vibrioses/imunologia , Vibrioses/microbiologia
20.
Fish Shellfish Immunol ; 59: 66-76, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27765699

RESUMO

Liver-expressed antimicrobial peptide 2 (LEAP-2) is a cationic peptide that plays an important role in the host's innate immune system. However, the mechanism by which LEAP-2 modulates/regulates the host defense against pathogens remains largely unknown. In this study, we identified a cDNA sequence encoding LEAP-2 homolog (BpLEAP-2) in the mudskipper, Boleophthalmus pectinirostris. Sequence analysis revealed that BpLEAP-2 belonged to the fish LEAP-2A cluster and that it was closely related to ayu LEAP-2. BpLEAP-2 mRNA was detected in a wide range of tissues, with the highest level of transcripts found in the liver. Upon infection with Edwardsiella tarda, BpLEAP-2 mRNA expression was significantly increased in the liver, kidney, spleen, and gill, but decreased in the intestine. Chemically synthesized BpLEAP-2 mature peptide did not exhibit antibacterial activity against E. tarda in vitro. Intraperitoneal injection of BpLEAP-2 (1.0 or 10.0 µg/g) resulted in significantly improved survival rate and reduced tissue bacterial load in E. tarda-infected mudskippers. In E. tarda-infected fish, BpLEAP-2 (0.1, 1.0, or 10.0 µg/g) eliminated E. tarda-induced tissue mRNA expression of BpTNF-α and BpIL-1ß. In monocytes/macrophages (MO/MФ), BpLEAP-2 (1.0 or 10.0 µg/ml) induced chemotaxis, enhanced respiratory burst, and inhibited E. tarda-induced mRNA expression of BpTNF-α and BpIL-1ß. At a concentration of 10.0 µg/ml, BpLEAP-2 also significantly enhanced the bacterial killing efficiency of MO/MФ. No significant effect was seen in the phagocytic activity of MO/MФ upon treatment with BpLEAP-2. Our study provides evidence, for the first time, that LEAP-2 exhibited immunomodulatory effects on immune cells, and protected the host from pathogenic infections independent of direct bacterial killing function.


Assuntos
Peptídeos Catiônicos Antimicrobianos/imunologia , Infecções por Enterobacteriaceae/veterinária , Doenças dos Peixes/imunologia , Peixes , Imunomodulação , Sequência de Aminoácidos , Animais , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/genética , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Edwardsiella tarda/fisiologia , Infecções por Enterobacteriaceae/imunologia , Infecções por Enterobacteriaceae/microbiologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Imunidade Inata/genética , Macrófagos/imunologia , Monócitos/imunologia , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Alinhamento de Sequência/veterinária
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