Developmental and comparative immunology

Publications in this journal

• Article: Identification of interferon- -inducible-lysosomal thiol reductase(GILT) gene from Mefugu (Takifugu obscures) and its immune response to LPS challenge.

  Meng Liu, Hongxin Ai, Wen Xiao, Yuefen Shen, Yang Shen, Xianwei Cui, Shuangquan Zhang
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  ABSTRACT: Interferon-γ-inducible-lysosomal thiol reductase (GILT) plays a key role in the processing and presentation of MHC class II-restricted antigen (Ag) by catalyzing disulfide bond reduction. In this study, a mefugu cDNA (ToGILT) encodes a deduced protein of 242 amino acids with a putative molecular weight of 28.6 kDa. It contains typical features of GILT proteins including the signature sequence CQHGX2ECX2NX4C, CXXC motif and other five cysteines. Genomic analysis revealed that ToGILT gene exhibited a similar exon-intron organization to human and mouse GILT. Phylogenetic analysis showed that ToGILT derived from a common ancestor with other vertebrate GILT proteins. The ToGILT mRNA was expressed in a tissue-specific manner and obviously up-regulated in spleen and kidney after LPS induction. These results suggest that ToGILT may be involved in the immune response to bacteria challenge in Takifugu obscurus .
  Developmental and comparative immunology 05/2013;
• Article: Overview of Drosophila immunity: a historical perspective.

  Jean-Luc Imler
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  ABSTRACT: The functional analysis of genes from the model organism Drosophila melanogaster has provided invaluable information for many cellular and developmental or physiological processes, including immunity. The best-understood aspect of Drosophila immunity is the inducible humoral response, first recognized in 1972. This pioneering work led to a remarkable series of findings over the next thirty years, ranging from the identification and characterization of the antimicrobial peptides produced, to the deciphering of the signalling pathways activating the genes that encode them and, ultimately, to the discovery of the receptors sensing infection. These studies on an insect model coincided with a revival of the field of innate immunity, and had an unanticipated impact on the biomedical field.
  Developmental and comparative immunology 05/2013;
• Article: Avian CD4(+)CD25(+) Regulatory T cells: Properties and Therapeutic Applications.

  Ramesh K Selvaraj
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  ABSTRACT: Regulatory T cells (Tregs) are a subset of T cells that specialize in immune suppression. CD4(+)CD25(+)FoxP3(+) T cells have been characterized as Tregs and extensively studied in mammals. In the absence of a putative FoxP3 ortholog in avians, CD4(+)CD25(+) cells is characterized as Tregs in avians. Avian CD4(+)CD25(+) cells produce high amounts of IL-10, TGF-β, CTLA-4, and LAG-3 mRNA; lack IL-2 mRNA; and suppress T cell proliferation in vitro through both contact-dependent and -independent pathways. Depleting avian CD4(+)CD25(+) cells increases the proliferation of, IL-2 amount, and IFNγ mRNA amount of CD4(+)CD25(-) cells. Avian CD4(+)CD25(+) cells lose their suppressive properties immediately after inflammation and acquire supersuppressive properties once inflammation subsides. Although Treg activity could be beneficial to the host, Tregs simultaneously inhibit host immunity and cause persistent infections of certain pathogens. Therapy targeted towards alleviating Treg mediated immune suppression can improve host immunity against those persistent pathogens and benefit poultry production.
  Developmental and comparative immunology 05/2013;
• Article: PmSERPIN3 from black tiger shrimp Penaeus monodon is capable of controlling the proPO system.

  Natthiya Wetsaphan, Vichien Rimphanitchayakit, Anchalee Tassanakajon, Kunlaya Somboonwiwat
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  ABSTRACT: Serpin or serine proteinase inhibitor is a family of protease inhibitors that are involved in controlling the proteolytic cascade in various biological processes. In shrimp, several serpins have been identified but only a few have been characterized. Herein, the PmSERPIN3 gene identified from Penaeus monodon EST database was studied. By using the 5'-and 3'-Rapid Amplification cDNA End (RACE) techniques, the full-length of PmSERPIN3 cDNA was obtained. The cDNA contained an open reading frame of 1,233 bp encoding for 410 amino acid residues protein. Genome sequence analysis revealed that the PmSERPIN3 was an intronless gene. RT-PCR analysis revealed that it was constitutively expressed in all developmental stages, all shrimp tissues tested, and upon pathogen infections. The recombinant mature PmSERPIN3 protein (rPmSERPIN3) produced in Escherichia coli exhibited inhibitory activity against subtilisin. The rPmSERPIN3 also inhibited the shrimp prophenoloxidase system activation in vitro. Injecting the rPmSERPIN3 along with Vibrio harveyi into the shrimp decreased the clearance rate of bacteria in the hemolymph. Potentially, the PmSERPIN3 functions as a regulator of the proPO activating system.
  Developmental and comparative immunology 05/2013;
• Article: Antigen Targeting to Apc: from Mice to Veterinary Species.

  B Alvarez, T Poderoso, F Alonso, A Ezquerra, J Domínguez, C Revilla
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  ABSTRACT: Antigen delivery to receptors expressed on antigen presenting cells (APC) has shown to improve immunogenicity of vaccines in mice. An enhancement of cytotoxic T lymphocyte (CTL), helper T cell or humoral responses was obtained depending on the type of APC and the surface molecule targeted. Although this strategy is being also evaluated in livestock animals with promising results, some discrepancies have been found between species and pathogens. The genetic diversity of livestock animals, the different pattern of expression of some receptors among species, the use of different markers to characterize APC in large animals and sometimes the lack of reagents make difficult to compare results obtained in different species. In this review, we summarize the data available regarding antigen targeting to APC receptors in cattle, sheep and pig and discuss the results found in these animals in the context of what has been obtained in mice.
  Developmental and comparative immunology 05/2013;
• Article: Immunity to bacterial infection in the chicken.

  Paul Wigley
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  ABSTRACT: Bacterial infections remain important to the poultry industry both in terms of animal and public health, the latter due to the importance of poultry as a source of foodborne bacterial zoonoses such as Salmonella and Campylobacter. As such, much focus of research to the immune response to bacterial infection has been to Salmonella. In this review we will focus on how research on avian salmonellosis has developed our understanding of immunity to bacteria in the chicken from understanding the role of TLRs in recognition of bacterial pathogens, through the role of heterophils, macrophages and γδ lymphocytes in innate immunity and activation of adaptive responses to the role of cellular and humoral immunity in immune clearance and protection. What is known of the immune response to other bacterial infections and in particular infections that have emerged recently as major problems in poultry production including Campylobacter jejuni, Avian Pathogenic Escherichia coli, Ornithobacterium rhinotracheale and Clostridium perfringens are discussed.
  Developmental and comparative immunology 05/2013;
• Article: Intracellular infections in Drosophila melanogaster: host defense and mechanisms of pathogenesis.

  Claire B Péan, Marc S Dionne
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  ABSTRACT: The fruit-fly Drosophila melanogaster has emerged as a powerful model to study innate immunity against intracellular pathogens. To combat infection, the fly relies on multiple lines of defense, many of which are shared with mammals and arthropod vectors of human diseases. In addition to conserved immune pathways, the ease of performing sophisticated genetic screens has allowed the identification of novel host immune factors and novel pathogen virulence factors. Recently, some groups have exploited this to simultaneously analyze the host and pathogen genetics of intracellular infection. This review aims to unravel the Drosophila immune response against intracellular pathogens, highlighting recent discoveries.
  Developmental and comparative immunology 05/2013;
• Article: Chicken immunoregulatory Ig-like receptor families: an overview and expression details on ggTREM-A1.

  Birgit C Viertlboeck, Matthias A Hanczaruk, Barbara Amann, Sophie R Bader, Ramona Schmitt, Beatrice Sperling, Susanne C N Schwarz, Wolfgang Schmahl, Cornelia A Deeg, Thomas W Göbel
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  ABSTRACT: Paired immunoregulatory receptors facilitate the coordination of the immune response at the cellular level. In recent years, our group characterized chicken homologues to mammalian immunoregulatory Ig-like receptor families. The first part of this review focuses on the current progress on chicken immunoregulatory Ig-like receptor families. One of these receptors is gallus gallus TREM-A1, which was described as the only member of the chicken TREM family with activating potential. The second part of this review presents a study initiated to further characterize ggTREM-A1 expression. For this purpose we established real-time RT-PCR and generated a specific mab to analyze the expression profile of ggTREM-A1 on mRNA and protein level, respectively. GgTREM-A1 mRNA was predominantly expressed in macrophages, but was also detected in brain, bone marrow, bursa, thymus, spleen and PBMC. Analyzing ggTREM-A1 surface expression by mab staining validated the expression on macrophages. Additionally, we showed high expression on blood monocytes, heterophils and NK cells and on monocytes isolated from bone marrow. Moreover, we detected ggTREM-A1 protein also on thrombocytes, B and T cell subsets, but antigen expression seemed to be lower and more variable in these cells. Immunohistochemistry of chicken brain tissue, combining ggTREM-A1 mab and various markers specific for various brain cell subsets showed expression of ggTREM-A1 on microglial cells, but also on neurons, astrocytes and oligodendrocytes. In conclusion, ggTREM-A1 is expressed on a variety of cells, relevant for the immune system, possibly combining physiological function of different mammalian TREM.
  Developmental and comparative immunology 05/2013;
• Article: Avian Host Defense Peptides.

  Tryntsje Cuperus, Maarten Coorens, Albert van Dijk, Henk P Haagsman
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  ABSTRACT: Host defense peptides (HDPs) are important effector molecules of the innate immune system of vertebrates. These antimicrobial peptides are also present in invertebrates, plants and fungi. HDPs display broad-spectrum antimicrobial activities and fulfill an important role in the first line of defense of many organisms. It is becoming increasingly clear that in the animal kingdom the functions of HDPs are not confined to direct antimicrobial actions. Research in mammals has indicated that HDPs have many immunomodulatory functions and are also involved in other physiological processes ranging from development to wound healing. During the past five years our knowledge about avian HDPs has increased considerably. This review addresses our current knowledge on the evolution, regulation and biological functions of HDPs of birds.
  Developmental and comparative immunology 05/2013;
• Article: Innate and adaptive immune mechanisms are effectively induced in ileal Peyer's patches of Salmonella typhimurium infected pigs.

  Rodrigo Prado Martins, Valentina Lorenzi, Cristina Arce, Concepción Lucena, Ana Carvajal, Juan José Garrido
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  ABSTRACT: In this report we employed laser-capture microdissection (LCM) coupled to qPCR technology and bioinformatic analysis to characterize, for the first time, the response of Peyer's patches (PP) from orally infected animals to S. typhimurium, in a model of non-typhoidal salmonellosis. Pathogen was highly found in the cytoplasm of phagocytes in PP and differential gene expression analysis indicated an up-regulation of proinflammatory molecules, establishment of a Th1 driven response and triggering of DC and T-cell activity. Furthermore, predictions by bioinformatic analysis pointed to an activation of processes regarding stimulation and maturation of DC, influx of leukocytes in tissue and T lymphocytes priming and differentiation. In short, the approach used in this study proved to be a promising strategy to explore infectious processes. Indeed, it revealed an effective induction of innate and adaptive immune mechanisms in swine PP which appear to be distinct from those observed in mesenteric lymph nodes and closely related to response of gut mucosa.
  Developmental and comparative immunology 05/2013;
• Article: Comparative genomic and phylogenetic analyses of the intelectin gene family: implications for their origin and evolution.

  Jie Yan, Lingxiao Xu, Yan Zhang, Changqing Zhang, Chao Zhang, Fei Zhao, Lijun Feng
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  ABSTRACT: Intelectin is a newly characterized gene family involved in early embryogenesis, host-pathogen interactions and iron metabolism. In this study, we searched the genomes of metazoans by extensive BLAST survey and found no intelectin homologs in invertebrate metazoans but 12 in amphioxus Branchiostoma floridae and 21 in ascidians Ciona intestinalis. Some ascidians oocyte cortical granule lectins (CGLs) have unknown insertion sequences between fibrinogen-related domain (FReD) and Intelectin Domain, the boundaries of which are equivalent to exon structures. In addition to ascidians intelectins/CGLs located in the base, phylogenetic tree comprises four main clades representing mammal, frog, fish, and amphioxus, indicating that intelectin genes undergo extensive lineage-specific duplication or gene conversion. However, genomic neighborhood surrounding analysis shows that clear proto-orthologies are difficult to be established among these counterparts. In addition, sequence comparison and phylogenetic analysis of FReDs from intelectins and other fibrinogen-like proteins from choanoflagellate, anemone, frog and human indicate FReDs of intelectins are unique. Likewise, these choanoflagellate and anemone genes may be close to intelectin gene.
  Developmental and comparative immunology 04/2013;
• Article: Conventional and Non-Conventional Drosophila Toll Signaling.

  Scott A Lindsay, Steven A Wasserman
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  ABSTRACT: The discovery of Toll in Drosophila and of the remarkable conservation in pathway composition and organization catalyzed a transformation in our understanding of innate immune recognition and response. At the center of that picture is a cascade of interactions in which specific microbial cues activate Toll receptors, which then transmit signals driving transcription factor nuclear localization and activity. Experiments gave substance to the vision of pattern recognition receptors, linked phenomena in development, gene regulation, and immunity into a coherent whole, and revealed a rich set of variations for identifying non-self and responding effectively. More recently, research in Drosophila has illuminated the positive and negative regulation of Toll activation, the organization of signaling events at and beneath membranes, the sorting of information flow, and the existence of non-conventional signaling via Toll-related receptors. Here, we provide an overview of the Toll pathway of flies and highlight these ongoing realms of research.
  Developmental and comparative immunology 04/2013;
• Article: Identification, expression profiling of a grass carp TLR8 and its inhibition leading to the resistance to reovirus in CIK cells.

  Xiaohui Chen, Qing Wang, Chunrong Yang, Youliang Rao, Qingmei Li, Quanyuan Wan, Limin Peng, Shuqin Wu, Jianguo Su
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  ABSTRACT: TLR8 (toll-like receptor 8), a homolog of TLR3, TLR7 and TLR9 as prototypical intracellular members of TLR family, is generally associated with sensing single stranded RNA and plays a pivotal role in antiviral immune response. In this study, a TLR8 gene from grass carp Ctenopharyngodon idella (designated as CiTLR8) was obtained and characterized. The full-length cDNA of CiTLR8 was of 3766 bp. The open reading frame was of 3072 bp and encoded a polypeptide of 1023 amino acids, including seventeen LRR (leucine-rich repeat) motifs, one transmembrane domain and one TIR (toll/interleukin-1 receptor) domain. A single intron with the size of 839 bp was found on the neck of start codon (ATG). CiTLR8 mRNA was ubiquitously expressed in the 15 tested tissues and the expression level in gas bladder, spleen, brain, hindgut and trunk kidney tissues was high. Besides, the CiTLR8 expression in spleen and head kidney was significantly up-regulated and reached peak at 24 h post-injection of grass carp reovirus (GCRV). CiTLR8 transcription reached peak at 8 h and then declined below the normal level post-GCRV infection in the C. idella kidney (CIK) cell line; and it was rapidly and significantly down-regulated by the stimulation of the synthetic double-stranded RNA polyriboinosinic-polyribocytidylic acid sodium salt (poly I:C) in CIK in a dose and time-dependent manner. The inhibitor expression vectors were constructed and transfected into CIK cell line to obtain stably expressing shRNA targeting TLR8. In CiTLR8-knockdown cells, CiTLR7 transcript weakly increased, CiIFN-I mRNA was significantly down-regulated, and the expression of CiMyD88, CiIRF7 and CiMx1 scarcely changed. Post poly I:C stimulation, CiTLR8, CiTLR7 and CiMyD88 transcripts significantly increased, CiIRF7 was down-regulated after an initial phase of increase, and CiIFN-I and CiMx1 transcripts were up-regulated. After GCRV infection, the transcripts of CiTLR8, CiTLR7, CiMyD88 and CiIRF7 were up-regulated, but CiIFN-I and CiMx1 transcripts were inhibited. Nevertheless, cells transfected with pshTLR8 vectors had strong resistance against GCRV injection, suggesting CiTLR8 might play a negative role in antiviral immune response. These results collectively suggested that CiTLR8 was a novel member of TLR gene family, engaging in antiviral innate immune defense in C. idella, which laid a foundation for the further mechanism research of TLR8 in fishes.
  Developmental and comparative immunology 04/2013;
• Article: Identification of cathepsin B in the razor clam Sinonovacula constricta and its role in innate immune responses.

  Donghong Niu, Kai Jin, Lie Wang, Fanyue Sun, Jiale Li
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  ABSTRACT: Cathepsin B, a lysosomal cysteine protease, has drawn much attention in vertebrates. However, very little is known about the functions of cathepsin B in bivalves. In this study, we identified the cathepsin B gene in the razor clam Sinonovacula constricta. The protein has a typical cysteine protease structure, comprising a 15-residue putative signal peptide, a 75-residue propeptide and a 249-residue mature domain. In the mature domain, there is an occluding loop, an oxyanion hole (Gln) and a catalytic triad (Cys, His and Asn). The cathepsin B gene is expressed in a wide range of tissues but appears to exhibit greatest level of expression in the liver. During the early developmental stages, the transcript could be detected widely. After the clam was infected with Vibrio anguillarum, the expression of the cathepsin B gene showed the most significant up-regulation in the liver and mantle tissues at 8 h after infection. The fact that bacterial infection can induce the expression of the cathepsin B transcript suggests that cathepsin B could play an important role in the innate immunity of clams.
  Developmental and comparative immunology 04/2013;
• Article: Unique Features of Chicken Toll-like Receptors.

  A Marijke Keestra, Marcel R de Zoete, Lieneke I Bouwman, Mahdi M Vaezirad, Jos P M van Putten
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  ABSTRACT: Toll-like receptors (TLRs) are a major class of innate immune pattern recognition receptors that have a key role in immune homeostasis and the defense against infections. The research explosion that followed the discovery of TLRs more than a decade ago has boosted fundamental knowledge on the function of the immune system and the resistance against disease, providing a rational for clinical modulation of the immune response. In addition, the conserved nature of the ancient TLR system throughout the animal kingdom has enabled a comparative biology approach to understand the evolution, structural architecture, and function of TLRs. In the present review we focus on TLR biology in the avian species, and, especially, on the unique functional properties of the chicken TLR repertoire.
  Developmental and comparative immunology 04/2013;
• Article: Immune Responses of Poultry to Newcastle Disease Virus.

  Darrell R Kapczynski, Claudio L Afonso, Patti J Miller
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  ABSTRACT: Newcastle disease (ND) remains a constant threat to poultry producers worldwide, in spite of the availability and global employment of ND vaccinations since the 1950s. Strains of Newcastle disease virus (NDV) belong to the order Mononegavirales, family Paramyxoviridae, and genus Avulavirus, are contained in one serotype and are also known as avian paramyxovirus serotype-1 (APMV-1). They are pleomorphic in shape and are single-stranded, non-segmented, negative sense RNA viruses. The virus has been reported to infect most orders of birds and thus has a wide host range. Isolates are characterized by virulence in chickens and the presence of basic amino acids at the fusion protein cleavage site. Low virulent NDV typically produce subclinical disease with some morbidity, whereas virulent isolates can result in rapid, high mortality of birds. Virulent NDV are listed pathogens that require immediate notification to the Office of International Epizootics and outbreaks typically result in trade embargos. Protection against NDV is through the use of vaccines generated with low virulent NDV strains. Immunity is derived from neutralizing antibodies formed against the viral hemagglutinin and fusion glycoproteins, which are responsible for attachment and spread of the virus. However, new techniques and technologies have also allowed for more in depth analysis of the innate and cell-mediated immunity of poultry to NDV. Gene profiling experiments have led to the discovery of novel host genes modulated immediately after infection. Differences in virus virulence alter host gene response patterns have been demonstrated. Furthermore, the timing and contributions of cell-mediated immune responses appear to decrease disease and transmission potential. In view of recent reports of vaccine failure from many countries on the ability of classical NDV vaccines to stop spread of disease, renewed interest in a more complete understanding of the global immune response of poultry to NDV will be critical to developing new control strategies and intervention programs for the future.
  Developmental and comparative immunology 04/2013;
• Article: Defense genes missing from the flight division.

  Katharine E Magor, Domingo Miranzo Navarro, Megan R W Barber, Kristina Petkau, Ximena Fleming-Canepa, Graham A D Blyth, Alysson H Blaine
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  ABSTRACT: Birds have a smaller repertoire of immune genes than mammals. In our efforts to study antiviral responses to influenza in avian hosts, we have noted key genes that appear to be missing. As a result, we speculate that birds have impaired detection of viruses and intracellular pathogens. Birds are missing TLR8, a detector for single-stranded RNA. Chickens also lack RIG-I, the intracellular detector for single-stranded viral RNA. Riplet, an activator for RIG-I, is also missing in chickens. IRF3, the nuclear activator of interferon-beta in the RIG-I pathway is missing in birds. Downstream of interferon signalling, some of the antiviral effectors are missing, including ISG15, and ISG54 and ISG56 (IFITs). Birds have only three antibody isotypes and IgD is missing. Ducks, but not chickens, make an unusual truncated IgY antibody that is missing the Fc fragment. Chickens have an expanded family of LILR leukocyte receptor genes, called CHIR genes, with hundreds of members, including several that encode IgY Fc receptors. Intriguingly, LILR homologues appear to be missing in ducks, including these IgY Fc receptors. The truncated IgY in ducks, and the duplicated IgY receptor genes in chickens may both have resulted from selective pressure by a pathogen on IgY FcR interactions. Birds have a minimal MHC, and the TAP transport and presentation of peptides on MHC class I is constrained, limiting function. In addition, ducks appear to lack tapasin, a chaperone involved in loading peptides on MHC class I. Finally, the absence of lymphotoxin-alpha and beta may account for the observed lack of lymph nodes in birds. Overall, the picture that emerges is some impairment of innate and adaptive immune responses in birds, especially to RNA viruses.
  Developmental and comparative immunology 04/2013;
• Article: Detection of the hepcidin prepropeptide and mature peptide in liver of rainbow trout.

  Claudio A Alvarez, Paula A Santana, Fanny Guzmán, Sergio Marshall, Luis A Mercado
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  ABSTRACT: Hepcidin is a small, cationic peptide which displays antimicrobial activities and iron regulatory function. Originally identified in mammals, this peptide is also present in fish. Hepcidin mRNA is predominantly expressed in liver and is regulated by iron and pathogen infection. In this work, we characterized the expression of trout hepcidin at protein level using rabbit antisera. Results showed that the prepropeptide of hepcidin can be detected by Western Blot in liver tissue from trout injected with iron or lipopolysaccharide. The mature hepcidin peptide was detected at the ionized state 5+ (m/z 577.2) by HPLC-ESI-MS in acid extracts from liver tissue. Moreover, hepcidin peptide was located in trout liver imprints by immunofluorescence. These results showed that hepcidin peptide is up-regulated by iron and bacterial components in the trout liver. This up-regulation could be a potential indicator of disease susceptibility, suggesting that hepcidin regulates iron homeostasis in salmonids.
  Developmental and comparative immunology 04/2013;
• Article: Molecular and cellular analyses of a ryanodine receptor from hemocytes of Pieres rapae.

  Shunfan Wu, Fei Wang, Jia Huang, Qi Fang, Zhicheng Shen, Gongyin Ye
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  ABSTRACT: Ryanodine receptors (RyRs) are located in the sarcoplasmic/endoplasmic reticulum membrane and are a distinct class of ligand-gated calcium channels controlling the release of calcium from intracellular stores. Intracellular calcium level has a definite role in innate and adaptive immune signaling. However, very few information are accessible about calcium transients of invertebrate immunocytes, especially of insect hemocytes, the effector cells of insect immunity. In this study, we show that the RyR-stimulating agent flubendiamide inhibit hemocyte spreading and phagocytosis in the cabbage white butterfly, Pieris rapae. Furthermore, we cloned a cDNA encoding a ryanodine receptor (PrRyR) from the hemocytes of P. rapae. It encodes 5107 amino acids with a predicted molecular weight of 578.2 kDa. PrRyR shares a common feature with known RyRs: a well-conserved COOH-terminal domain with two consensus calcium-binding EF-hands and six transmembrane domains, and a large hydrophilic NH2-terminal domain. In the larval stage, PrRyR was highly expressed in epidermis tissue and also expressed in hemocytes at a moderate level. In the adult stage, PrRyR was expressed at high levels in thoraces and legs, while low levels in abdomens and antennae. Quantitative real-time PCR analysis showed that its expression did not display any significant change in response to bacterial challenge. Western blot analysis and immunohistochemistry assay displayed that PrRyR was detected and presented on hemocytes. We also showed that flubendiamide, a RyR-activating insecticide, induced Ca(2+) release and thereby confirmed functional expression of the PrRyR in the hemocytes of P. rapae.
  Developmental and comparative immunology 04/2013;
• Article: Genomic characterization and transcriptional evidence for the involvement of complement component 7 in immune response of rock bream (Oplegnathus fasciatus).

  W D Niroshana Wickramaarachchi, Ilson Whang, Eunmi Kim, Bong-Soo Lim, Hyung-Bok Jeong, Mahanama De Zoysa, Myung-Joo Oh, Sung-Ju Jung, Sang-Yeob Yeo, Sung Yeon Kim, Hae-Chul Park, Jehee Lee
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  ABSTRACT: The complement component 7 (C7) is the central mediator of pathogenic attack at the membrane surface and its binding to the C5b-7 complex triggers cytolytic signaling. In this study, C7 of rock bream (Oplegnathus fasciatus) was identified (Rb-C7) and characterized at the genomic level. The Rb-C7 gene contains 18 exons and 17 introns and is composed of a 2490 bp complete open reading frame (ORF). The encoded polypeptide (830 amino acids) contains a number of well-conserved C7 signature domains. Important putative transcription factor binding sites, including those for NF-κB, SP-1, C/EBP, AP-1 and OCT-1, are present in the 5'-flanking region of Rb-C7. Phylogenetic analysis revealed a close proximity of Rb-C7 with the orthologues in tilapia and Japanese flounder. Quantitative real-time PCR (qPCR) analysis confirmed constitutive Rb-C7 expression throughout all the examined tissue of healthy rock bream, with highest expression in liver. In immune challenge experiment, Rb-C7 expression was up-regulated in head kidney and liver in response to Edwardsiella tarda, Streptococcus iniae, lipopolysaccharide and rock bream iridovirus (RBIV). Furthermore, significant increases of both intracellular expression level and the number of Rb-C7-expressing cells were detected by in situ hybridization assay in head kidney and liver tissues upon E. tarda infection. These results suggested that Rb-C7 is lytic pathway gene in complement system and its transcriptional regulation may be an important immune response in pathogenic defense mechanism of rock bream.
  Developmental and comparative immunology 04/2013;