To counteract host immunity, Cryptosporidium parvum has evolved multiple strategies to suppress host antimicrobial defense. One such strategy is to reduce the production of the antimicrobial peptide beta-defensin 1 (DEFB1) by host epithelial cells but the underlying mechanisms remain unclear. Recent studies demonstrate that a panel of parasite RNA transcripts of low protein-coding potential are delivered into infected host cells and may modulate host gene transcription. Using in vitro models of intestinal cryptosporidiosis, in this study, we analyzed the expression profile of host beta-defensin genes in host cells following infection. We found that C. parvum infection caused a significant downregulation of the DEFB1 gene. Interestingly, downregulation of DEFB1 gene was associated with host delivery of Cdg7_FLc_1000 RNA transcript, a C. parvum RNA that has previously demonstrated to be delivered into the nuclei of infected host cells. Knockdown of Cdg7_FLc_1000 in host cells could attenuate the trans-suppression of host DEFB1 gene and decreased the parasite burden. Therefore, our data suggest that trans-suppression of DEFB1 gene in intestinal epithelial cells following C. parvum infection involves host delivery of parasite Cdg7_FLc_1000 RNA, a process that may be relevant to the epithelial defense evasion by C. parvum at the early stage of infection.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Abrahamsen MS, Templeton TJ, Enomoto S, Abrahante JE, Zhu G, Lancto CA, Deng M, Liu C, Widmer G, Tzipori S, Buck GA, Xu P, Bankier AT, Dear PH, Konfortov BA, Spriggs HF, Iyer L, Anantharaman V, Aravind L, Kapur V (2004) Complete genome sequence of the apicomplexan, Cryptosporidium parvum. Science 304(5669):441–445. https://doi.org/10.1126/science.1094786
Carmen JC, Sinai AP (2007) Suicide prevention: disruption of apoptotic pathways by protozoan parasites. Mol Microbiol 64(4):904–916. https://doi.org/10.1111/j.1365-2958.2007.05714.x
Carmen JC, Hardi L, Sinai AP (2006) Toxoplasma gondii inhibits ultraviolet light-induced apoptosis through multiple interactions with the mitochondrion-dependent programmed cell death pathway. Cell Microbiol 8:301–315
Checkley W, White AC, Jaganath D, Arrowood MJ, Chalmers RM, Chen XM, Fayer R, Griffiths JK, Guerrant RL, Hedstrom L, Huston CD, Kotloff KL, Kang G, Mead JR, Miller M, Petri WA Jr, Priest JW, Roos DS, Striepen B, Thompson RC, Ward HD, Van Voorhis WA, Xiao L, Zhu G, Houpt ER (2015) A review of the global burden, novel diagnostics, therapeutics, and vaccine targets for Cryptosporidium. Lancet Infect Dis 15(1):85–94. https://doi.org/10.1016/S1473-3099(14)70772-8
Chen XM, LaRusso NF (2000) Mechanisms of attachment and internalization of Cryptosporidium parvum to biliary and intestinal epithelial cells. Gastroenterology 118(2):368–379. https://doi.org/10.1016/S0016-5085(00)70219-8
Chen XM, Levine SA, Splinter PL, Tietz PS, Ganong AL, Jobin C, Gores GJ, Paya CV, LaRusso NF (2001) Cryptosporidium parvum activates nuclear factor kappaB in biliary epithelia preventing epithelial cell apoptosis. Gastroenterology 120(7):1774–1783. https://doi.org/10.1053/gast.2001.24850
Chen XM, Keithly JS, Paya CV, LaRusso NF (2002) Cryptosporidiosis. N Engl J Med 346(22):1723–1731. https://doi.org/10.1056/NEJMra013170
Chen XM, O’Hara SP, Nelson JB, Splinter PL, Small AJ, Tietz PS, Limper AH, LaRusso NF (2005) Multiple toll-like receptors are expressed in human cholangiocytes and mediate host epithelial responses to C. parvum via activation of NF-kappaB. J Immunol 175(11):7447–7456. https://doi.org/10.4049/jimmunol.175.11.7447
Deng M, Lancto CA, Abrahamsen MS (2004) Cryptosporidium parvum regulation of human epithelial cell gene expression. Int J Parasitol 34(1):73–82. https://doi.org/10.1016/j.ijpara.2003.10.001
Hu G, Gong AY, Roth AL, Huang BQ, Ward HD, Zhu G, Larusso NF, Hanson ND, Chen XM (2013) Release of luminal exosomes contributes to TLR4-mediated epithelial antimicrobial defense. PLoS Pathog 9(4):e1003261. https://doi.org/10.1371/journal.ppat.1003261
Jarczak J, Kościuczuk EM, Lisowski P, Strzałkowska N, Jóźwik A, Horbańczuk J, Krzyżewski J, Zwierzchowski L, Bagnicka E (2013) Defensins: natural component of human innate immunity. Hum Immunol 74(9):1069–1079. https://doi.org/10.1016/j.humimm.2013.05.008
Kotloff KL, Nataro JP, Blackwelder WC, Nasrin D, Farag TH, Panchalingam S, Wu Y, Sow SO, Sur D, Breiman RF, Faruque AS, Zaidi AK, Saha D, Alonso PL, Tamboura B, Sanogo D, Onwuchekwa U, Manna B, Ramamurthy T, Kanungo S, Ochieng JB, Omore R, Oundo JO, Hossain A, Das SK, Ahmed S, Qureshi S, Quadri F, Adegbola RA, Antonio M, Hossain MJ, Akinsola A, Mandomando I, Nhampossa T, Acácio S, Biswas K, O'Reilly CE, Mintz ED, Berkeley LY, Muhsen K, Sommerfelt H, Robins-Browne RM, Levine MM (2013) Burden and aetiology of diarrhoeal disease in infants and young children in developing countries (the Global Enteric Multicenter Study, GEMS): a prospective, case-control study. Lancet 382(9888):209–222. https://doi.org/10.1016/S0140-6736(13)60844-2
Lantier L, Lacroix-Lamandé S, Potiron L, Metton C, Drouet F, Guesdon W, Gnahoui-David A, Le Vern Y, Deriaud E, Fenis A, Rabot S, Descamps A, Werts C, Laurent F (2013) Intestinal CD103+ dendritic cells are key players in the innate immune control of Cryptosporidium parvum infection in neonatal mice. PLoS Pathog 9(12):e1003801. https://doi.org/10.1371/journal.ppat.1003801
Laurent F, Kagnoff MF, Savidge TC, Naciri M, Eckmann L (1998) Human intestinal epithelial cells respond to Cryptosporidium parvum infection with increased prostaglandin H synthase 2 expression and prostaglandin E2 and F2alpha production. Infect Immun 66(4):1787–1790
Liao Q, Shen J, Liu J, Sun X, Zhao G, Chang Y, Xu L, Li X, Zhao Y, Zheng H, Zhao Y, Wu Z (2014) Genome-wide identification and functional annotation of Plasmodium falciparum long noncoding RNAs from RNA-seq data. Parasitol Res 113(4):1269–1281. https://doi.org/10.1007/s00436-014-3765-4
Liu J, Deng M, Lancto CA, Abrahamsen MS, Rutherford MS, Enomoto S (2009) Biphasic modulation of apoptotic pathways in C. parvum-infected intestinal epithelial cells. Infect Immun 77(2):837–849. https://doi.org/10.1128/IAI.00955-08
McDonald V, Korbel DS, Barakat FM, Choudhry N, Petry F (2013) Innate immune responses against Cryptosporidium parvum infection. Parasite Immunol 35(2):55–64. https://doi.org/10.1111/pim.12020
Ming ZP, Gong AY, Wang Y, Zhang XT, Li M, Mathy NW, Strauss-Soukup JK, Chen XM (2018) Involvement of Cryptosporidium parvum Cdg7_FLc_1000 RNA in the attenuation of intestinal epithelial cell migration via trans-suppression of host cell SMPD3 gene. J Infect Dis 217:122–133
O’Donoghue PJ (1995) Cryptosporidium and cryptosporidiosis in man and animals. Int J Parasitol 25(2):139–195. https://doi.org/10.1016/0020-7519(94)E0059-V
O'Connor RM, Wanyiri JW, Wojczyk BS, Kim K, Ward H (2007) Stable expression of Cryptosporidium parvum glycoprotein gp40/15 in Toxoplasma gondii. Mol Biochem Parasitol 152(2):149–158. https://doi.org/10.1016/j.molbiopara.2007.01.003
Petersen CA, Krumholz KA, Carmen J, Sinai AP, Burleigh BA (2006) Trypanosoma cruzi infection and nuclear factor kappa B activation prevent apoptosis in cardiac cells. Infect Immun 74(3):1580–1587. https://doi.org/10.1128/IAI.74.3.1580-1587.2006
Prasanth KV, Spector DL (2007) Eukaryotic regulatory RNAs: an answer to the ‘genome complexity’ conundrum. Genes Dev 21(1):11–42. https://doi.org/10.1101/gad.1484207
Puiu D, Enomoto S, Buck GA, Abrahamsen MS, Kissinger JC (2004) CryptoDB: the Cryptosporidium genome resource. Nucleic Acids Res 32:329–331
Rogers KA, Rogers AB, Leav BA, Sanchez A, Vannier E, Uematsu S, Akira S, Golenbock D, Ward HD (2006) MyD88-dependent pathways mediate resistance to Cryptosporidium parvum infection in mice. Infect Immun 74(1):549–556. https://doi.org/10.1128/IAI.74.1.549-556.2006
Schmidt W, Wahnschaffe U, Schäfer M, Zippel T, Arvand M, Meyerhans A, Riecken EO, Ullrich R (2001) Rapid increase of mucosal CD4 T cells followed by clearance of intestinal cryptosporidiosis in an AIDS patient receiving highly active antiretroviral therapy. Gastroenterology 120(4):984–987. https://doi.org/10.1053/gast.2001.22557
Semple F, Dorin JR (2012) β-Defensins: multifunctional modulators of infection, inflammation and more? J Innate Immun 4(4):337–348. https://doi.org/10.1159/000336619
Sibley LD (2004) Intracellular parasite invasion strategies. Science 304(5668):248–253. https://doi.org/10.1126/science.1094717
Striepen B (2013) Parasitic infections: time to tackle cryptosporidiosis. Nature 503(7475):189–191. https://doi.org/10.1038/503189a
Ulitsky I, Bartel DP (2013) lincRNAs: genomics, evolution, and mechanisms. Cell 154(1):26–46. https://doi.org/10.1016/j.cell.2013.06.020
Vembar SS, Scherf A, Siegel TN (2014) Noncoding RNAs as emerging regulators of Plasmodium falciparum virulence gene expression. Curr Opin Microbiol 20:153–161
Vinayak S, Pawlowic MC, Sateriale A, Brooks CF, Studstill CJ, Bar-Peled Y, Cipriano MJ, Striepen B (2015) Genetic modification of the diarrhoeal pathogen Cryptosporidium parvum. Nature 523(7561):477–480. https://doi.org/10.1038/nature14651
Wang Y, Gong AY, Ma S, Chen X, Li Y, Su CJ, Norall D, Chen J, Strauss-Soukup JK, Chen XM (2017a) Delivery of parasite RNA transcripts into infected epithelial cells during Cryptosporidium infection and its potential impact on host gene transcription. J Infect Dis 215(4):636–643. https://doi.org/10.1093/infdis/jiw607
Wang Y, Gong AY, Ma S, Chen X, Strauss-Soukup JK, Chen XM (2017b) Delivery of parasite Cdg7_Flc_0990 RNA transcript into intestinal epithelial cells during Cryptosporidium parvum infection suppresses host cell gene transcription through epigenetic mechanisms. Cell Microbiol 19(11):e12760. https://doi.org/10.1111/cmi.12760
Yamagishi J, Wakaguri H, Sugano S, Kawano S, Fujisaki K, Sugimoto C, Watanabe J, Suzuki Y, Kimata I, Xuan X (2011) Construction and analysis of full-length cDNA library of Cryptosporidium parvum. Parasitol Int 60(2):199–202. https://doi.org/10.1016/j.parint.2011.03.001
Yang YL, Serrano MG, Sheoran AS, Manque PA, Buck GA, Widmer G (2009) Over-expression and localization of a host protein on the membrane of Cryptosporidium parvum infected epithelial cells. Mol Biochem Parasitol 168(1):95–101. https://doi.org/10.1016/j.molbiopara.2009.07.004
Zaalouk TK, Bajaj-Elliott M, George JT, McDonald V (2004) Differential regulation of beta-defensin gene expression during Cryptosporidium parvum infection. Infect Immun 72(5):2772–2779. https://doi.org/10.1128/IAI.72.5.2772-2779.2004
Zhou R, Hu G, Liu J, Gong AY, Drescher KM, Chen XM (2009) NF-kappaB p65-dependent transactivation of miRNA genes following Cryptosporidium parvum infection stimulates epithelial cell immune responses. PLoS Pathog 5(12):e1000681. https://doi.org/10.1371/journal.ppat.1000681
Zhou R, Gong AY, Eischeid AN, Chen XM (2012) miR-27b targets KSRP to coordinate TLR4-mediated epithelial defense against Cryptosporidium parvum infection. PLoS Pathog 8(5):e1002702. https://doi.org/10.1371/journal.ppat.1002702
We thank Dr. Guanghui Zhao (College of Veterinary Medicine, Northwest A&F University) for helpful and stimulating discussions, and Barbara L. Bittner (Creighton University) for her assistance in writing the manuscript. This work was supported by funding from the National Institutes of Health (AI116323 and AI136877) and the Nebraska Stem Cell Research Program (LB606), and by revenue from Nebraska’s excise tax on cigarettes awarded to Creighton University through the Nebraska Department of Health & Human Services (DHHS) (LB595). Dr. Zhenping Ming was a visiting scholar supported by the China Scholarship Council and the National Natural Science Foundation of China (NSFC No. 31372194). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health, the State of Nebraska, DHHS or NSFC.
Conflict of interest
The authors declare that they have no conflict of interest.
Section Editor: Lihua Xiao
About this article
Cite this article
Ming, Z., Gong, A., Wang, Y. et al. Trans-suppression of defense DEFB1 gene in intestinal epithelial cells following Cryptosporidium parvum infection is associated with host delivery of parasite Cdg7_FLc_1000 RNA. Parasitol Res 117, 831–840 (2018). https://doi.org/10.1007/s00436-018-5759-0
- Intestinal epithelium
- Gene transcription
- Epithelial defense