Identification and characterization of common carp (Cyprinus carpio L.) granzyme A/K, a cytotoxic cell granule-associated serine protease

Huang, Rong; Zhong, Shan; Liu, Hong; Kong, Renqiu; Wang, Yaping; Hu, Wei; Guo, Qinghai

doi:10.1016/j.fsi.2010.04.002

Cited by 19 publications

(12 citation statements)

References 50 publications

(53 reference statements)

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“…Both gzma transcription and/or tryptase/GzmA/K activity are increased in HKLs during CMC assays against NNV-infected target cells and up-regulated upon in vivo NNV infection in both fish species. Similarly, mRNA levels of gzma are also up-regulated in fish by viral infections: in common carp ( Cyprinus carpio ) by spring viremia of carp virus (SVCV) (17), in rainbow trout ( Oncorhynchus mykiss ) RTS-11 cell line and leucocytes by viral haemorrhagic septicaemia virus (VHSV) (43), in Atlantic salmon ( Salmo salar ) by infectious pancreatic necrotic virus (IPNV) (44, 45) or in sea bass retina by NNV (46). In addition, mRNA levels of Atlantic salmon and ginbuna crucian carp gzma were also up-regulated by bacterial infections (24, 45).…”

Section: Discussionmentioning

confidence: 99%

Fish Granzyme A Shows a Greater Role Than Granzyme B in Fish Innate Cell-Mediated Cytotoxicity

et al. 2019

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Granzymes (Gzm) are serine proteases, contained into the secretory granules of cytotoxic cells, responsible for the cell-mediated cytotoxicity (CMC) against tumor cells and intracellular pathogens such as virus and bacteria. In fish, they have received little attention to their existence, classification or functional characterization. Therefore, we aimed to identify and evaluate their functional and transcriptomic relevance in the innate CMC activity of two relevant teleost fish species, gilthead seabream and European sea bass. Afterwards, we wanted to focus on their regulation upon nodavirus (NNV) infection, a virus that causes great mortalities to sea bass specimens while seabream is resistant. In this study, we have identified genes encoding GzmA and GzmB in both seabream and sea bass, as well as GzmM in seabream, which showed good phylogenetic relation to their mammalian orthologs. In addition, we found enzymatic activity related to tryptase (GzmA and/or GzmK), aspartase (GzmB), metase (GzmM), or chymase (GzmH) in resting head-kidney leucocytes (HKLs), with the following order of activity: GzmA/K ~ GzmM >> GzmH >>> GzmB. In addition, during innate CMC assays consisting on HKLs exposed to either mock- or NNV-infected target cells, though all the granzyme transcripts were increased only the tryptase activity did. Thus, our data suggest a high functional activity of GzmA/K in the innate CMC and a marginal one for GzmB. Moreover, GzmB activity was detected into target cells during the CMC assays. However, the percentage of target cells with GzmB activity after the CMC assays was about 10-fold lower than the death target cells, demonstrating that GzmB is not the main inductor of cell death. Moreover, in in vivo infection with NNV, gzm transcription is differently regulated depending on the fish species, genes and tissues. However, the immunohistochemistry study revealed an increased number of GzmB stained cells and areas in the brain of seabream after NNV infection, which was mainly associated with the lesions detected. Further studies are needed to ascertain the molecular nature, biological function and implication of fish granzymes in the CMC activity, and in the antiviral defense in particular.

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Section: Discussionmentioning

confidence: 99%

Fish Granzyme A Shows a Greater Role Than Granzyme B in Fish Innate Cell-Mediated Cytotoxicity

et al. 2019

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“…Of the remaining five host defence proteins, granzyme A/K is a member of the serine protease family (Huang et al . ) along with natural killer cell enhancing factor, which is a component of natural kill cells, and macrophage migration inhibitory factor (MIF) is a member of the cytokine signalling pathway (Bernhagen et al . ).…”

Section: Discussionmentioning

confidence: 99%

“…In addition to the C3-H1 protein, three other proteins (complements C3-S, C4-1 and C4-2) were identified in the complement host defence pathway (Kato et al 2004). Of the remaining five host defence proteins, granzyme A/K is a member of the serine protease family (Huang et al 2010) along with natural killer cell enhancing factor, which is a component of natural kill cells, and macrophage migration inhibitory factor (MIF) is a member of the cytokine signalling pathway (Bernhagen et al 1993). Belcourt et al (1995) demonstrated that granulin-1 localizes to macrophages in common carp and gold fish, Carassius auratus auratus, and granulin-3 which was identified in the gel analysis also is likely part of the host defence pathway.…”

Section: Discussionmentioning

confidence: 99%

Antibody screening identifies 78 putative host proteins involved in Cyprinid herpesvirus 3 infection or propagation in common carp, Cyprinus carpio L

Gotesman

Soliman

El-Matbouli

2013

Journal of Fish Diseases

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Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of a serious and notifiable disease afflicting common and koi carp, Cyprinus carpio L., termed koi herpesvirus disease (KHVD). Significant progress has been achieved in the last 15 years, since the initial reports surfaced from Germany, USA and Israel of the CyHV-3 virus, in terms of pathology and detection. However, relatively few studies have been carried out in understanding viral replication and propagation. Antibody-based affinity has been used for detection of CyHV-3 in enzyme-linked immunosorbent assay and PCR-based techniques, and immunohistological assays have been used to describe a CyHV-3 membrane protein, termed ORF81. In this study, monoclonal antibodies linked to Nhydroxysuccinimide (NHS)-activated spin columns were used to purify CyHV-3 and host proteins from tissue samples originating in either CyHV-3 symptomatic or asymptomatic fish. The samples were next analysed either by polyacrylamide gel electrophoresis (PAGE) and subsequently by electrospray ionization coupled to mass spectrometry (ESI-MS) or by ESI-MS analysis directly after purification. A total of 78 host proteins and five CyHV-3 proteins were identified in the two analyses. These data can be used to develop novel control methods for CyHV-3, based on pathways or proteins identified in this study.

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“…Perforin gene expression is usually up-regulated after immunization of ginbuna crucian carp with tumor cells (Toda et al, 2011a), after PHA-L (Phaseolus vulgaris leucoagglutinin) stimulation of trout CD8 + cells (Takizawa et al, 2011), after VHSV infection of RTS11 cell line (Ordás et al, 2011) and after viral infection or DNA vaccination in rainbow trout (unpublished data), whilst down-regulated after cadmium exposure (Auslander et al, 2008). In a similar fashion, granzyme genes are up-regulated by bacterial vaccination (Caipang et al, 2008), viral infections (VHSV in RTS11 cell line and SVCV in carp) (Huang et al, 2010;Ordás et al, 2011) and viral infection or DNA vaccination (unpublished data). The transcript level of CD8 gene is related to the CTL presence, abundance and activity.…”

Section: Modulation Of the Fish Cytotoxic Activitymentioning

confidence: 91%

“…The release of perforin and granzyme contained in the granules is calcium-dependent and the NCC activity is greatly inhibited or completely abrogated by Ca 2+ -chelators demonstrating their involvement in the NCCmediated cytotoxic activity (Carlson et al, 1985;Hogan et al, 1999). In the last decade, fish perforin (Athanasopoulou et al, 2009;Hwang et al, 2004;Toda et al, 2011a) and granzyme (Huang et al, 2010;Praveen et al, 2004Praveen et al, , 2006Wernersson et al, 2006) sequences have been obtained but their gene expression or function has been scarcely related to the innate cytotoxic activity (Ordás et al, 2011;Praveen et al, 2006). The granule-independent killing mechanism has also been identified in fish NCCs by the use of commercial antibodies or functional studies (Ca-chelators) leading to the identification of the Fas/FasL system in fish NCCs (Bishop et al, 2002;Cuesta et al, 2003a;Evans et al, 200, 2001;Jaso-Friedmann et al, 2000;Kaur et al, 2004;Long et al, 2004).…”

Section: Lymphocytesmentioning

confidence: 99%

Fighting Virus and Parasites with Fish Cytotoxic Cells

Esteban¹,

Meseguer²,

Cuesta³

2012

Health and Environment in Aquaculture

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Identification and characterization of common carp (Cyprinus carpio L.) granzyme A/K, a cytotoxic cell granule-associated serine protease

Cited by 19 publications

References 50 publications

Fish Granzyme A Shows a Greater Role Than Granzyme B in Fish Innate Cell-Mediated Cytotoxicity

Fish Granzyme A Shows a Greater Role Than Granzyme B in Fish Innate Cell-Mediated Cytotoxicity

Antibody screening identifies 78 putative host proteins involved in Cyprinid herpesvirus 3 infection or propagation in common carp, Cyprinus carpio L

Fighting Virus and Parasites with Fish Cytotoxic Cells

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