Regulation of transcription of the Aedes albopictus cecropin A1 gene: A role for p38 mitogen-activated protein kinase

Moon, Alice; Walker, Anthony J.; Goodbourn, Stephen

doi:10.1016/j.ibmb.2011.04.001

Cited by 15 publications

(12 citation statements)

References 51 publications

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“…In contrast to Drosophila , deletion of GATA sites did not affect B. mori CecA1 promoter activity, and this despensible role of GATA site was also be identified in BmCecropinB1 and A. albopictus Cecropin A1 promoter (Moon et al ., ). In addition, the F2BPI‐binding site which consists of two CATTA and is necessary for full activation of BmCeccropinB1 appeared to be nonfunctional at all since deletion of one CATTA upstream of κB motif had no effect on promoter activity, and the proximal CATTA cannot sustain the promoter activity in the absence of κB motif, suggesting that no other factors independent of NF‐κB contributed to the inducibility of CecA1 promoter.…”

Section: Discussionmentioning

confidence: 97%

Characterization of the Bombyx mori Cecropin A1 promoter regulated by IMD pathway

Hua

Xue

et al. 2015

Insect Science

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Cecropin A1 (CecA1) promoter from Bombyx mori was cloned and characterized to provide insight into the transcriptional control of this antimicrobial peptide gene upon immune challenges. Reporter gene assays demonstrated that both Escherichia coli and lipopolysaccharide could induce expression in BmE cells but B. bombyseptieus or peptidoglycan failed, and the induction pattern of the reporter gene was coincident with the endogenous CecA1. Analysis of deletion and mutation constructs revealed that the regulatory region was the κB motif located between -176 and -166, and no other predicted elements on CecA1 promoter affected its inducibility. Insertion of additional κB motifs increased the activity of CecA1 promoter. Furthermore, binding of Relish to κB motif was confirmed by electrophoretic mobility shift assay. These findings indicate the regulatory mechanism of CecA1 expression in IMD pathway and suggest an approach of engineering antimicrobial peptide promoter with enhanced activities that may lead to broad applications.

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Section: Discussionmentioning

confidence: 97%

Characterization of the Bombyx mori Cecropin A1 promoter regulated by IMD pathway

Hua

Xue

et al. 2015

Insect Science

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“…gambiae (5.1* and Sua5B), Lutzomya longipalpis (LL5 cells), Ae. albopictus (especially C6/36 cells), among other insects, have long been used to investigate different aspects of insect immunity, being essential tools for the construction of the knowledge we have today regarding Toll, IMD, Jak/STAT and RNAi pathways in insects [17-24]. …”

Section: Introductionmentioning

confidence: 99%

“…albopictus larvae homogenates by Singh [28]. C6/36 has been used to understand the regulation of the synthesis and secretion of several important proteins/processes, such as defensin [29], cecropin [24] or phagocytosis [19] but mostly this cell has been used to study aspects of insect-virus relations [17]. This is because one of the main features of C6/36 is the capacity of growing a very broad spectrum of viruses and producing higher virus titers than any other cell [30].…”

Section: Introductionmentioning

confidence: 99%

Validation of Aedes aegypti Aag-2 cells as a model for insect immune studies

Barletta

Silva²,

Sorgine

2012

Parasites Vectors

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BackgroundThe understanding of mosquito immune responses can provide valuable tools for development of novel mosquito control strategies. Aiming the study at insect innate immunity, continuous insect cell lines have been established and used as research tools due to the fact that they constitute more homogeneous, sensitive, and reproducible systems than the insects from which they originated. More recently, Aag-2, an Aedes aegypti cell lineage, began to be frequently used as a model for studies of mosquito immunity. Nevertheless, to our knowledge, no study has systematically characterized the responses of Aag-2 cell line against different kinds of pathogens and compared its response to those exhibited by whole mosquitoes. For this reason, in this study we characterized gene expression profiles of the Aag-2 cell line in response to different kinds of immune challenges, such as Gram negative and positive bacteria, fungi and viruses, comparing the obtained results with the ones already described in the literature for whole mosquitoes.MethodsAedes aegypti Aag-2 cells were exposed to different immune stimuli (gram-positive and gram negative heat inactivated bacteria, zymosan or Sindbis virus) for 24 hours and the expression of selected marker genes from toll, IMD and Jak/STAT pathways was analyzed by qPCR. Also, cells were incubated with fluorescent latex beads for evaluation of its phagocytosis capacity.ResultsAag-2 cells were stimulated with two concentrations of heat-killed Gram negative (Enterobacter cloacae) or Gram positive (Micrococcus luteus) bacteria, Zymosan or infected with Sindbis virus and the expression of key genes from the main immune related pathways, Toll, IMD and Jak/STAT, were investigated. Our results suggest that Toll and IMD pathways are activated in response to both Gram positive and negative bacteria and Zymosan in Aag-2 cells, displaying an immune profile similar to those described in the literature for whole mosquitoes. The same stimuli were also capable of activating Jak/STAT pathway in Aag-2 cells. Infection with Sindbis virus led to an up-regulation of the transcription factor STAT but was not able to induce the expression of any other gene from any of the pathways assayed. We also showed that this cell line is able to phagocytose latex beads in culture.ConclusionsOur results characterize the expression profile of Aag-2 cells in response to different immune stimuli and demonstrate that this cell lineage is immune-competent and closely resembles the response described for whole Ae. aegypti mosquitoes. Hence, our findings support the use of Aag-2 as a tool to comprehend Ae. aegypti immune response both at cellular and humoral levels.

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“…) and cecropin (Moon et al . ). Mostly, however, this cell line has been used to study aspects of vector–virus relationships (Sanchez‐Vargas et al .…”

Section: Discussionmentioning

confidence: 97%

“…Most studies that have focused on mosquito immune responses against pathogens have employed the lineage C6/36 (Igarashi 1978;Miller & Brown 1992;Adelman et al 2002;Sanchez-Vargas et al 2004). The C6/36 cell line has been used to demonstrate regulation of the synthesis and secretion of several important proteins, such as defensin (Cheng et al 2001) and cecropin (Moon et al 2011). Mostly, however, this cell line has been used to study aspects of vector-virus relationships (Sanchez-Vargas et al 2004).…”

Section: Defensin a Transcript And Peptide Expression In C6/36 Infectionmentioning

confidence: 99%

Induction of defensin response to dengue infection in Aedes aegypti

Wasinpiyamongkol

Missé

Luplertlop

2014

Entomological Science

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Innate immune‐related defensin peptide expression, the major antimicrobial peptide (AMP) in the dengue vector mosquito Aedes aegypti, was analyzed following infection by dengue virus type 2 (strain 16681) (DENV‐2). In vitro, the mosquito cell line C6/36 was exposed to dengue virus at different multiplicities of infection (MOI) in an hour; it was found that the expression level of defensin transcripts was dependent upon viral dose. In addition, using both polymerase chain reaction (PCR) and Western blotting assays to determine defensin transcription and translation at a series of times post‐inoculation, we found that the time course of defensin transcripts correlated with the expression of defensin peptide post‐infection. In vitro, female Ae. aegypti mosquitoes were fed different meals (DENV‐2 infected blood, non‐infectious blood and sugar) at 14 days post‐feeding, to determine defensin transcription in response to dengue infection, by surface‐enhanced laser desorption/ionization time‐of‐flight mass spectrometry (SELDI‐TOF‐MS) assay. The results showed that a peak of 4.25 kDa peptide, composed of defensin, was predominantly induced by DENV‐2 infected blood‐fed mosquitoes. We observed that DENV‐2 could trigger the defensin peptide, defined molecularly as an innate immune response. Therefore, this peptide may be involved in dengue infection and/or transmission. Improved understanding of the mosquito's responses to dengue virus should strengthen our understanding of this vector's innate immune system.

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Regulation of transcription of the Aedes albopictus cecropin A1 gene: A role for p38 mitogen-activated protein kinase

Cited by 15 publications

References 51 publications

Characterization of the Bombyx mori Cecropin A1 promoter regulated by IMD pathway

Characterization of the Bombyx mori Cecropin A1 promoter regulated by IMD pathway

Validation of Aedes aegypti Aag-2 cells as a model for insect immune studies

Induction of defensin response to dengue infection in Aedes aegypti

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