De Bosscher, Karolien

Dreesen, Leentje

Geldhof, Peter

Li, RW

Paerewijck, Oonagh

Ratman, Dariusz

Van Meulder, Frederik

English

Ghent University Academic Bibliography

Authors	   O.	  Paerewijck1,	  L.	  Dreesen1,	  F.	  Van	  Meulder1,	  D.	  Ratman2,	  K.	  De	  Bosscher2,	  R.W.	  Li3,	  P.	  Geldhof1	  Affiliations	   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	  3Agriculture	   Research	   Service,	   Animal	   Genomics	   and	   Improvement	   Laboratory,	  Beltsville,	  Maryland,	  United	  States	  of	  America	  Title	   Unraveling	   the	   role	   of	   IL-­‐17A	   in	   the	   intestinal	   immune	   response	   against	   the	  protozoan	  parasite	  Giardia	  muris	  by	  an	  RNA	  sequencing	  approach	  	   Background	  The	   intestinal	  protozoan	  parasite	  Giardia	  duodenalis	  has	  a	  wide	  vertebrate	  host	  range.	   	   An	   infection	   with	   Giardia	   can	   lead	   to	   gastro-­‐intestinal	   complaints.	  Although	  in	  most	  hosts	  these	  symptoms	  rapidly	  disappear,	  a	  chronic	  situation	  can	  develop.	   Recent	   data	   obtained	   in	   mice,	   cattle	   and	   humans	   indicates	   that	  development	  of	   immunity	   is	  dependent	  on	   IL-­‐17A.	  The	  aim	  of	   this	   study	  was	   to	  further	   unravel	   the	   IL-­‐17A	   induced	   effector	   mechanisms	   in	   a	   Giardia-­‐mouse	  infection	  model.	  Methods	  C57Bl/6	   wildtype	   (WT)	   and	   IL-­‐17RA-­‐KO	   mice	   were	   orally	   infected	   with	   103	  G.	  muris	  cysts.	  Three	  weeks	  post-­‐infection,	   intestinal	  tissue	  samples	  were	  collected	  for	   RNA	   purification	   and	   subsequent	   transcriptome	   analysis.	   Samples	   collected	  from	   non-­‐infected	   WT	   and	   KO	   animals	   served	   as	   negative	   controls.	   Genes	   of	  interest	   were	   further	   subjected	   to	   transcription	   and	   expression	   analysis	   and	  immunolocalisation.	   Their	   functional	   role	   was	   investigated	   by	   performing	  infection	  experiments	  in	  KO	  mice,	  pending	  availability.	  	  Results	  Transcriptome	   analysis	   indicated	   that	   844	   genes	   were	   differentially	   expressed	  between	   WT	   infected	   and	   WT	   control	   mice,	   many	   of	   which	   have	   never	   been	  linked	   to	   Giardia	   infection	   before,	   including	   genes	   associated	   with	   circadian	  rhythm.	  Comparison	  of	  WT	  infected	  with	  IL-­‐17RA-­‐KO	  infected	  mice	  resulted	  in	  the	  identification	   of	   287	   differentially	   transcribed	   genes.	   One	   of	   these,	   mannose-­‐binding	   lectin	   2,	   is	   particularly	   interesting	   as	   it	   is	   involved	   in	   complement	  activation.	   Further	   work	   indicated	   that	   the	   expression	   and	   secretion	   of	   MBL2	  following	  a	  Giardia	   infection	   is	   IL-­‐17A	  dependent	  and	   infection	   studies	   in	  Mbl2-­‐KO	  mice	  showed	  a	  higher	  load	  of	  Giardia	  trophozoites	  in	  comparison	  to	  WT	  mice.	  	  Conclusions	  The	  transcriptome	  approach	  that	  was	  followed	  resulted	  in	  the	  identification	  of	  a	  number	   of	   genes	   that	   seem	   to	   play	   a	   role	   in	   the	   IL-­‐17A	   response	   following	   a	  Giardia	   infection.	   Additional	   research	   is	   now	   ongoing	   to	   further	   unravel	   the	  involvement	  of	  these	  genes	  in	  the	  development	  of	  intestinal	  immunity.	  	  	  

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

Paerewijck, OonaghUGent0000706791498020016415589739261787670000-0002-8272-0559096077CA-F0EE-11E1-A9DE-61C894A0A6B4

Dreesen, LeentjeUGent00200304312880200063680297829900272802F72A5A-F0EE-11E1-A9DE-61C894A0A6B4

Van Meulder, Frederik00007074856480200095882197791520646431BCB29C-F0EE-11E1-A9DE-61C894A0A6B4

Ratman, DariuszUGent0000814191708020005879959748581526461C262D1E-F0EE-11E1-A9DE-61C894A0A6B4

De Bosscher, KarolienGE318010010331090000-0001-5059-9718F4EF58C4-F0ED-11E1-A9DE-61C894A0A6B4

Geldhof, PeterDI048010012358960000-0001-8222-2966F57157B6-F0ED-11E1-A9DE-61C894A0A6B4

Archivsystem Ask23

 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

https://biblio.ugent.be/publication/7278001/file/7278002.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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Ghent University Academic Bibliography

Archivsystem Ask23

Ghent University Academic Bibliography