De Bosscher, Karolien

Dreesen, Leentje

Geldhof, Peter

Li, RW

Paerewijck, Oonagh

Ratman, Dariusz

Van Meulder, Frederik

English

Ghent University Academic Bibliography

 The aim of the study was to unravel the effector mechanisms related to the protective IL-17A response against G. muris in mice. This research is financed by the Research Foundation - Flanders  C57Bl/6 WT and C57Bl/6 IL-17RA KO mice were orally infected with 103 G. muris cysts. 2 groups of respective non-infected control mice were included in the study. The protozoan parasite Giardia duodenalis is a highly common intestinal pathogen with a wide vertebrate host range, including humans. Giardia muris is a natural parasite of rodents. Giardia species have a simple and direct life cycle. After infective cysts are ingested orally, excystation occurs, resulting in the release of flagellated trophozoites that attach to the mucosa of the small intestine. After some rounds of asexual reproduction, encystation takes place. The cysts are then shed with the faeces, ready to be taken up by a new host.  An infection with Giardia is in many cases acute and self-limiting. However, a significant proportion of infected hosts develop a chronic infection. In mice infected with G. muris, clearing of the parasite is achieved around week 3 post-infection. Transcriptional analysis of small intestinal tissue revealed strong upregulation of IL-17A after infection. Furthermore, 3 weeks post-infection, IL-17A receptor A KO mice were unable to mount a protective immune response against G. muris.! There is a very dinstinct gene expression pattern in the small intestine of G. muris infected C57Bl/6 WT mice versus IL-17RA KO mice, reflected by 287 differentially expressed genes. In IL-17RA KO mice that fail to mount a protective immune response against the parasite, there is evidence for ongoing tissue damage 3 weeks post-infection, instead of tissue repair.   3 weeks post-infection the intestinal transcriptome of the 4 groups of mice was analysed by RNA-seq. Differentially expressed genes in the 4 comparisons were identified with DESeq2 software and used as an input for Ingenuity Pathway Analysis (IPA) software in order to extract biological pathways and functions. Genes were plotted in heatmaps to visualize dominant expression patterns.   Paerewijck Oonagh1,  Dreesen L.1, Van Meulder F.1, Ratman D.2, De Bosscher K.2,  Li R.W.3, Geldhof P.1 1Department of Virology, Parasitology and Immunology, Laboratory of Parasitology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium 2VIB Department of Medical Protein Research, Ghent University, Ghent, Belgium 3United States Department of Agriculture, Agriculture Research Service, Animal Genomics and Improvement Laboratory, Beltsville, Maryland, United States of America Corresponding author: Oonagh.Paerewijck@UGent.be   WT Infected    WT Control  KO Infected    KO Control  Unraveling the role of IL-17A in the intestinal immune response against the protozoan parasite Giardia muris by an RNA sequencing approach   Table 1. Top 5 of the most impacted and increased functions as predicted by IPA in the comparisons WT infected versus WT control and KO infected versus KO control A heatmap was created with 140 genes that are responsive to infection in either genotype, but that are differentially expressed between WT and KO mice upon infection. A second heatmap provides an overview of 31 differentially expressed genes between WT control versus KO control mice, that differentially respond upon infection in WT versus KO  mice.     Pathway analysis revealed the involvement of pathways linked to tissue damage in KO mice and pathways involved in tissue repair in WT mice. Figure 1. 844 genes were differentially expressed between WT infected and WT control mice 153 genes between KO infected and KO control mice 287 genes between WT infected and KO infected mice 415 genes between WT control and KO control mice   HEATMAP 1 HEATMAP 2 KO INF vs KO CT             153 WT INF vs KO INF              287 WT_CtrlWT_InfKO_CtrlKO_Inf Ly6g6dAA467197Prg2Mid1Klrb1bClec1bKlrb1cEtv4Klra3Cxcl5Shroom1PigrMe1Abcc3Nox1Angpt1Mfap5Clec9aBarx2Dock2Cd8b1Gm3893Zbtb16Sgk1SiglechAmotl1Pck1BC064078Gm11166Cecr6Bcl2l15−1 0 1Row Z−ScoreColor KeyWT_CtrlWT_InfKO_CtrlKO_InfCyp2j6BcheAkr1c19Olfr165Gpr1511100001G20RikKlra5AfmReg3bApol11bReg3gOas1cAcadlGm156Reg3aAldh1a1Slc6a20aDdit4lCd7Apol10aPsmb9Fscn3Cpn2Car2PtprcapOas3Gm12250IgtpGcsamTap1Irf7Trim30aGzmaRasd1Cxcl9Nkg7Il2rbWdpcpSellHavcr1ThpoZbp1Oas1aOlfr56Herc6Oas1gCcl5Ifi44Oasl2Phf11dGzmbIfi47Gbp6Bst2Oas2Gbp10Dusp2Slamf1Serpina3fIl18bpTgtp1Il10Mybl1Tox2Tgtp2Cd6A430093F15RikCd5Slc13a2osIl17aDppa3AvilNmuSh2d7Mbl2RetnlgAlox5Nfe2Ccdc129Alox5apItga2bEdn1Emilin2Gfi1bGsdmc2GsdmcStrip2Rgs22Frmpd1Clca3BC048546Gm973Tmem27Pou2f3Gp1baOptcCldn20Reg4Lama3Rgs13Trpc3Slc4a11Defa17ChatRgs18Ly6kTal1Kcnj16Ang4Actc1Mrgpra9A4gntNos2Csf2rb2Gpr97CpvlGata2Cd177Rab44Cd34Gpr123Cd300lbDuox2Adcy8Krt12Crybb3Tnfsf15CemipVgfGmprSpink4Sucnr1Gnat3Gm15299Defa−ps1KlAgr3Gm5475Tmem229aAcox2−1 0 1Row Z−ScoreColor KeyHeatmap 2. Heatmap of genes that are differentially expressed between WT control versus KO control mice and that differentially respond upon infection in WT infected versus KO infected mice Polymeric immunoglobulin receptor  Transporter protein able to carry large quantities of IgA across the intestinal epithelium  WT INF vs KO INF Fold change 2,05 WT_CtrlWT_InfKO_CtrlKO_InfCyp2j6BcheAkr1c19Olfr165Gpr1511100001G20RikKlra5AfmReg3bApol11bReg3gOas1cAcadlGm156Reg3aAldh1a1Slc6a20aDdit4lCd7Apol10aPsmb9Fscn3Cpn2Car2PtprcapOas3Gm12250IgtpGcsamTap1Irf7Trim30aGzmaRasd1Cxcl9Nkg7Il2rbWdpcpSellHavcr1ThpoZbp1Oas1aOlfr56Herc6Oas1gCcl5Ifi44Oasl2Phf11dGzmbIfi47Gbp6Bst2Oas2Gbp10Dusp2Slamf1Serpina3fIl18bpTgtp1Il10Mybl1Tox2Tgtp2Cd6A430093F15RikCd5Slc13a2osIl17aDppa3AvilNmuSh2d7Mbl2RetnlgAlox5Nfe2Ccdc129Alox5apItga2bEdn1Emilin2Gfi1bGsdmc2GsdmcStrip2Rgs22Frmpd1Clca3BC048546Gm973Tmem27Pou2f3Gp1baOptcCldn20Reg4Lama3Rgs13Trpc3Slc4a11Defa17ChatRgs18Ly6kTal1Kcnj16Ang4Actc1Mrgpra9A4gntNos2Csf2rb2Gpr97CpvlGata2Cd177Rab44Cd34Gpr123Cd300lbDuox2Adcy8Krt12Crybb3Tnfsf15CemipVgfGmprSpink4Sucnr1Gnat3Gm15299Defa−ps1KlAgr3Gm5475Tmem229aAcox2−1 0 1Row Z−ScoreColor KeyHeatmap 1. Heatmap of genes that differentially respond upon infection in WT infected mice versus KO infected mice Angiogenin 4 Paneth-cell derived anti-microbial peptide important in epithelial host defense in the small intestine  WT INF vs KO INF Fold change 4,35  Mannose-binding lectin 2 Humoral pattern-recognition molecule with a role in innate immunity, activates the complement system  WT INF vs KO INF Fold change 20,37  Materials and Methods Aim of the study Introduction Results Conclusions WT INF vs WT CT   Vasoconstriction    Development of cardiovascular system   Vasculogenesis   Development of blood vessel   Development of epithelial tissue KO INF vs KO CT   Apoptosis of neurons   Quantity of anion   Apoptosis of brain cells   Damage of kidney   Blood pressure WT_CtrlWT_InfKO_CtrlKO_Inf Cyp2j6BcheAkr1c19Olfr165Gpr1511100001G20RikKlra5AfmReg3bApol11bReg3gOas1cAcadlGm156Reg3aAldh1a1Slc6a20aDdit4lCd7Apol10aPsmb9Fscn3Cpn2Car2PtprcapOas3Gm12250IgtpGcsamTap1Irf7Trim30aGzmaRasd1Cxcl9Nkg7Il2rbWdpcpSellHavcr1ThpoZbp1Oas1aOlfr56Herc6Oas1gCcl5Ifi44Oasl2Phf11dGzmbIfi47Gbp6Bst2Oas2Gbp10Dusp2Slamf1Serpina3fIl18bpTgtp1Il10Mybl1Tox2Tgtp2Cd6A430093F15RikCd5Slc13a2osIl17aDppa3AvilNmuSh2d7Mbl2RetnlgAlox5Nfe2Ccdc129Alox5apItga2bEdn1Emilin2Gfi1bGsdmc2GsdmcStrip2Rgs22Frmpd1Clca3BC048546Gm973Tmem27Pou2f3Gp1baOptcCldn20Reg4Lama3Rgs13Trpc3Slc4a11Defa17ChatRgs18Ly6kTal1Kcnj16Ang4Actc1Mrgpra9A4gntNos2Csf2rb2Gpr97CpvlGata2Cd177Rab44Cd34Gpr123Cd300lbDuox2Adcy8Krt12Crybb3Tnfsf15CemipVgfGmprSpink4Sucnr1Gnat3Gm15299Defa−ps1KlAgr3Gm5475Tmem229aAcox2−1 0 1Row Z−ScoreColor KeyWT_CtrlWT_InfKO_CtrlKO_InfLy6g6dA467197Prg2Mid1Klrb1bClec1bKlrb1cEtv4Klra3Cxcl5Shroom1PigrMe1Abcc3Nox1Angpt1Mfap5Clec9aBarx2Dock2Cd8b1Gm3893Zbtb16Sgk1SiglechAmotl1Pck1BC064078Gm11166Cecr6Bcl l15−1 0 1Row Z−ScoreColor Key

Unraveling the role of IL-17A in the intestinal immune response against the protozoan parasite Giardia muris by an RNA sequencing approach

https://biblio.ugent.be/publication/7277840/file/7277920.pdf

Unraveling the role of IL-17A in the intestinal immune response against the protozoan parasite Giardia muris by an RNA sequencing approach

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