A Proteomic Approach To Investigate the Distribution and Abundance of Surface and Internal<i>Fasciola hepatica</i>Proteins during the Chronic Stage of Natural Liver Fluke Infection in Cattle

Haçarız, Orçun; Sayers, G.; Baykal, Ahmet Tarık

doi:10.1021/pr300015p

Cited by 47 publications

(62 citation statements)

References 72 publications

(139 reference statements)

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“…Previously, we have isolated F. hepatica tegumental coat Ag (FhTeg) and examined its interactions with innate immune cells. Most of the isolated proteins from FhTeg (21)(22)(23) and the contents of the glycocalyx (24) are identified. More than 50 different proteins are present in the preparation, and the biological active component or components of FhTeg have yet to be identified.…”

Section: Ast Cells Are Associated With Helminth Infection Butmentioning

confidence: 99%

Fasciola hepatica Tegumental Coat Impairs Mast Cells’ Ability To Drive Th1 Immune Responses

Vukman

Adams

Metz

et al. 2013

The Journal of Immunology

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The parasitic worm Fasciola hepatica induces strong Th2 and T-regulatory immune responses while simultaneously suppressing Th1-driven immune responses to bystander microbial infections. It also prevents the initiation of Th1-mediated autoimmune disorders in mice through the suppression of Th17 and Th1 immune responses, and this can be mimicked by parasite-derived molecules. We have isolated F. hepatica tegumental coat Ag (FhTeg) and demonstrated its suppressive effect in vivo by directly targeting dendritic cells, impairing their ability to drive Th1 responses. Mast cells are critical in promoting Th1 protective immunity during bacterial infection and in driving Th1-mediated pathological conditions in autoimmune diseases. In this article, we show that FhTeg inhibits the ability of mast cells to drive the Th1 immune response by suppressing cytokine secretion (TNF-α, IL-6, IFN-γ, and IL-10) and ICAM1 expression in mast cells stimulated with LPS or heat-inactivated Bordetella pertussis Ag. These heat-inactivated B. pertussis Ag/LPS–stimulated mast cells fail to promote Th1 immune responses in CD4+ T cells when pretreated with FhTeg, and a role for ICAM1 in this process was demonstrated. FhTeg suppresses the activation of transcription factors in the TLR signaling pathway, which explains the decrease in cytokine production and cell surface marker expression. We demonstrated that FhTeg suppresses MAPK and NF-κB activation and enhances SOCS3 expression, which could explain its negative effect on the TLR pathways. We conclude that FhTeg targets innate immune cells, inhibiting their ability to drive Th1 immune responses.

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Section: Ast Cells Are Associated With Helminth Infection Butmentioning

confidence: 99%

Fasciola hepatica Tegumental Coat Impairs Mast Cells’ Ability To Drive Th1 Immune Responses

Vukman

Adams

Metz

et al. 2013

The Journal of Immunology

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“…Sample preparation was done according to an earlier published protocol (27). Cultured cells were collected with HyQtase (Hyclone, SV3003001) treatment and later washed twice with 50 mM ice-cold ammonium bicarbonate.…”

Section: Sample Preparationmentioning

confidence: 99%

“…The parameters for the LC-MS analysis were optimized previously (27). A tryptic peptide mixture was spiked with 50 fmol MassPREP Enolase Digestion Standard (Waters Corp.).…”

Section: Lc-ms/ms Analysismentioning

confidence: 99%

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Proteomic evidence for the plasticity of cultured vascular smooth muscle cells

Baykal¹,

Baykal²,

Serhatlı³

et al. 2013

Turk J Biol

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Morphological, functional, and gene expression studies have established the phenotypic plasticity of smooth muscle cells (SMCs). These cells have been shown to respond to environmental stimulants such as extracellular matrix and growth factors. Cellular changes can vary between extremes defined as the contractile and synthetic states. Various growth factors have been shown to have profound effects on the phenotype of these cells. In this study, we intended to investigate the effects of growth factor-rich medium on the protein expression of vascular SMCs in culture. Interestingly, transiently changing the type of medium did not result in any apparent morphological differences, yet we hypothesized that some cellular factors might still be altered. In order to understand what kind of intracellular molecular changes should be expected to occur during the medium change, we analyzed global protein expression changes using nano-LC-MS/MS in smooth muscle cell cultures that were isolated and grown in one medium formulation and temporarily switched to the other. Our data indicate that proteins playing a role in energy metabolism (glycolysis), translation, and folding of proteins are affected, along with regulatory molecules and cytoskeletal proteins. The individual proteins and their significance are discussed within the scope of this paper.

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“…Parazit-konakçı ilişkisinin iyi bir şekilde anlaşılması, aşı veya daha etkili ilaç uygulamaları geliştirmek için çalışmalar yapılmaktadır (5,6). Genel olarak, deneysel çalışmalarda, F. hepatica parazitleri son konakçının karaciğer kanallarından çıkarıldıktan sonra FTS (fosfat tamponlu su) içerisine bırakılmaktadır.…”

Section: Introductionunclassified

Investigation of the Abundance of Proteins Secreted by Fasciola hepatica, Which is Exposed to Environmental Change in Experimental Studies, with an Advanced Proteomic Approach

Haçarız¹,

Baykal²

2014

urkiyeParazitolDerg

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ÖZETAmaç: Ana konakçıdan çıkarıldıktan sonra çevresel değişikliğe maruz kalan Fasciola hepatica'nın salgıladığı protein miktarlarını bir ileri proteomik yaklaşımla incelemek. Yöntemler: Son konakçıdan toplanan ergin F. hepatica parazitleri doğrudan fosfat tamponlu su (FTS, oda sıcaklığında) içerisine konuldu ve 37°C'de 2 saat bekletildi (Enstitü'ye 1 saat içerisinde getirildikten sonra). Daha sonra, FTS, bir revize F. hepatica protein veri bankası (Universal Protein Resource; UniProt) ve veri bağımsız edinim yöntemi ile ileri bir proteomik yöntem (yüksek başarımlı sıvı kromatografisine bağlı, elektriksel püskürtme ile iyonlaştırma ve dört kutup zaman bazlı ölçüm içeren kütle spektrometresi sistemi; nanoUPLC-ESI-QTOF-MS) kullanılarak incelendi. Bulgular: Parazitlerle bekletilme sonrası FTS'nin proteomik analizinde, cathepsin L protease 1, fatty acid binding protein 1 ve 2, thioredoxin peroxidase (TPx), ve kunitz type proteinase inhibitor tanımlandı. Fasciola hepatica TPx miktarı, bu çalışmada tanımlanan diğer proteinlerin miktarlarına göre yaklaşık olarak 2-6 kat daha fazla bulundu (p<0,01). Sonuç: Çevresel değişiklikten kaynaklanan parazit üzerindeki stres, TPx salınımının uyarılması ile ilişkili olabilir. İleri proteomik yaklaşımların uygulanması, parazit-konakçı etkileşiminin aydınlatılmasında ve parazite karşı etkili korunma yöntemlerinin geliştirilmesinde yararlı veriler sağlayabilir. (Turkiye Parazitol Derg 2014; 38: 106-10) Anahtar Sözcükler: Fasciola hepatica, TPx, proteomik, çevresel değişiklik, stres Geliş Tarihi: 16.11.2013Kabul Tarihi: 24.01.2014ABSTRACT Objective: Investigation of the abundance of proteins secreted by Fasciola hepatica, which is exposed to environmental change after it is removed from the main host, with an advanced proteomic approach. Methods: Adult Fasciola hepatica parasites, obtained from the main host, were directly placed in phosphate-buffered saline (PBS, at room temperature) and incubated at 37°C for 2 hours (after arrival at the Institute within 1 hour). After this, without applying extra procedures, such as washing the parasites, secreted parasite proteins in PBS were investigated using an advanced proteomic method [a mass spectrometry system with electrospray ionization and quadrupole time-of-flight source coupled to ultra performance liquid chromatography, nano UPLC-ESI-QTOF-MS] with a reviewed F. hepatica protein database (Universal Protein Resource; UniProt) and data-independent acquisition method. ), pozitif iyon V modu, MS ve MS/MS fonksiyonları (1,5 saniye aralıklarla 6V düşük enerji ve 15-40V yüksek enerji) kullanılarak pepResults: With the proteomic analysis of the PBS, after incubation with the parasites, cathepsin L protease 1, fatty acid-binding protein 1 and 2, thioredoxin peroxidase (TPx), and kunitz-type proteinase inhibitor were identified. The abundance of Fasciola hepatica TPx was approximately 2-6 times higher than that of the other proteins identified in this study (p<0.01). Conclusion: The stress on the parasite stem from environmental change could b...

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A Proteomic Approach To Investigate the Distribution and Abundance of Surface and InternalFasciola hepaticaProteins during the Chronic Stage of Natural Liver Fluke Infection in Cattle

Cited by 47 publications

References 72 publications

Fasciola hepatica Tegumental Coat Impairs Mast Cells’ Ability To Drive Th1 Immune Responses

Fasciola hepatica Tegumental Coat Impairs Mast Cells’ Ability To Drive Th1 Immune Responses

Proteomic evidence for the plasticity of cultured vascular smooth muscle cells

Investigation of the Abundance of Proteins Secreted by Fasciola hepatica, Which is Exposed to Environmental Change in Experimental Studies, with an Advanced Proteomic Approach

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