Cryptosporidium parvum alters glucose transport mechanisms in infected enterocytes
- 135 Downloads
The parasite Cryptosporidium parvum Tyzzer 1912 destroys parts of the intestinal brush border membrane which is important for the uptake of nutrients like glucose. In this study, glucose transport mechanisms of the host cells (IPEC-J2 cells) infected by C. parvum were investigated. The mRNA expression levels of glucose transporters (GLUT) 1 and 2 and Na+-coupled glucose transporter (SGLT) 1 were compared in infected and uninfected cells over an infection time of 24–96 h by RT-qPCR. Furthermore, the protein expression of SGLT 1 and GLUT 2 was quantified in western blot studies. While the protein expression of SGLT 1 was not altered in infected cells, mRNA expression of SGLT 1 and GLUT 1 was significantly increased 24 h p. i. and decreased 96 h p. i. The mRNA expression of GLUT 2 was significantly decreased 24 h, 72 h, and 96 h p. i. and also correlated significantly with the infection dose at 72 h p. i. In contrast to that, the protein expression of GLUT 2 was significantly increased 48 h p. i., associated with a significantly higher intracellular glucose level in infected cells compared with control cells at that time point of infection. This points to an adaptation of the host cells’ glucose uptake taking place in the acute phase of the infection. A better understanding of these molecular mechanisms following a C. parvum infection may probably lead to an improvement of therapy strategies in the future.
KeywordsC. parvum infection Glucose transport IPEC-J2 cells Parasite-host interaction
The authors want to thank S. Gawlowska and I. Urbansky for excellent technical assistance.
This study was funded by a starting grant of the Faculty of Veterinary Medicine, University of Leipzig as well as the “Freundeskreis Tiermedizin der Veterinärmedizinischen Fakultät Leipzig e.V.” The Leica TCS SP8 laser scanning microscope as well as the graphic work station equipped with IMARIS 9.3 was provided by the BioImaging Core Facility, University of Leipzig.
Compliance with ethical standards
Conflict of interest statement
The authors declare that they have no conflict of interest.
Statement of informed consent
All applicable international, national, and/or institutional guidelines for the care and use of the animals were followed. All procedures concerning the animals, which were used for the passage of the parasite, were performed in accordance with the ethical standards of the institution (Regional Council of Saxony following German law: TierSchG, TieSchVersV; permit number: A 06/19).
- Berschneider HM (1989) Development of normal cultured small intestinal epithelial cell lines which transport Na and Cl (Abstract). Gasteroenterology 96:A41Google Scholar
- Bürzle M, Hediger MA (2012) Functional and physiological role of vitamin C transporters. Curr Top Membr 70:357–375. https://doi.org/10.1016/B978-0-12-394316-3.00011-9 CrossRefPubMedGoogle Scholar
- Dengler F, Rackwitz R, Pfannkuche H, Gäbel G (2018) Coping with hypoxia: adaptation of glucose transport mechanisms across equine jejunum epithelium. JEVS 69:1–10Google Scholar
- Dominguez JH, Song B, Maianu L, Garvey WT, Qulali M (1994) Gene expression of epithelial glucose transporters: the role of diabetes mellitus. J Am Soc Nephrol. 5:29–36Google Scholar
- Ferguson SH, Foster DM, Sherry B, Magness ST, Nielsen DM, Gookin JL (2019) Interferon-λ3 promotes epithelial defense and barrier function against Cryptosporidium parvum infection. Cell Mol Gastroenterol Hepatol 8:1–20. https://doi.org/10.1016/j.jcmgh.2019.02.007 CrossRefPubMedPubMedCentralGoogle Scholar
- Foster DM, Stauffer SH, Stone MR, Gookin JL (2012) Proteasome inhibition of pathologic shedding of enterocytes to defend barrier function requires X-linked inhibitor of apoptosis protein and nuclear factor κB. Gastroenterology 143:133–144. https://doi.org/10.1053/j.gastro.2012.03.030 CrossRefPubMedGoogle Scholar
- Göhring F, Möller-Holtkamp P, Daugschies A, Lendner M (2014) Untersuchungen zur Häufigkeit von Cryptosporidium parvum bei Durchfallkälbern und der Einfluss von Koinfektionen auf das Krankheitsgeschehen. Tierärztl Umschau 69:000–000Google Scholar
- Kellett GL, Brot-Laroche E, Mace OJ, Leturque A (2008) Sugar absorption in the intestine: the role of GLUT2. Annu Rev Nutr 28:35–54. https://doi.org/10.1146/annurev.nutr.28.061807.155518 CrossRefPubMedGoogle Scholar
- 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 382:209–222CrossRefGoogle Scholar
- Lendner M, Etzold M, Daugschies A (2011) Kryptosporidiose – ein Update. Berl Münch Tierärztl Wochenschr 124:10–21Google Scholar
- Long W, Cheeseman CI (2015) Cell Health Cytoskeleton 7:167-83Google Scholar
- Mirhashemi ME, Noubary F, Chapman-Bonofiglio S, Tzipori S, Huggins GS, Widmer G (2018) Transcriptome analysis of pig intestinal cell monolayers infected with Cryptosporidium parvum asexual stages. Parasit Vectors 11:176. https://doi.org/10.1186/s13071-018-2754-3 CrossRefPubMedPubMedCentralGoogle Scholar
- Notari L, Riera DC, Sun R, Bohl JA, McLean LP, Madden KB, van Rooijen N, Vanuytsel T, Urban JF Jr, Zhao A, Shea-Donohue T (2014) Role of macrophages in the altered epithelial function during a type 2 immune response induced by enteric nematode infection. PLoS One 9:e84763. https://doi.org/10.1371/journal.pone.0084763 eCollection 2014CrossRefPubMedPubMedCentralGoogle Scholar
- O’Hara SP, Gajdos GB, Trussoni CE, Splinter PL, LaRusso NF (2010) Cholangiocyte myosin IIB is required for localized aggregation of sodium glucose cotransporter 1 to sites of Cryptosporidium parvum cellular invasion and facilitates parasite internalization. Infect Immun 78:2927–2936. https://doi.org/10.1128/IAI.00077-10 CrossRefPubMedPubMedCentralGoogle Scholar
- Platts-Mills JA, Babji S, Bodhidatta L, Gratz J, Haque R, Havt A, McCormick BJ, McGrath M, Olortegui MP, Samie A, Shakoor S, Mondal D, Lima IF, Hariraju D, Rayamajhi BB, Qureshi S, Kabir F, Yori PP, Mufamadi B, Amour C, Carreon JD, Richard SA, Lang D, Bessong P, Mduma E, Ahmed T, Lima AA, Mason CJ, Zaidi AK, Bhutta ZA, Kosek M, Guerrant RL, Gottlieb M, Miller M, Kang G, Houpt ER (2015) MAL-ED Network Investigators. Pathogen – specific burdens of community diarrhoea in developing countries: a multisite birth cohort study (MAL-ED). Lancet Glob Health 3:e564–e575CrossRefGoogle Scholar
- Schierack P, Nordhoff M, Pollmann M, Weyrauch KD, Amasheh S, Lodemann U, Jores J, Tachu B, Kleta S, Blikslager A, Tedin K, Wieler LH (2006) Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine. Histochem Cell Biol 125:293–305CrossRefGoogle Scholar
- Sekikawa S, Kawai Y, Fujiwara A, Takeda K, Tegoshi T, Uchikawa R, Yamada M, Arizono N (2003) Alterations in hexose, amino acid and peptide transporter expression in intestinal epithelial cells during Nippostrongylus brasiliensis infection in the rat. Int J Parasitol 33 (12):1419–1426CrossRefGoogle Scholar
- Steinhoff-Wagner J, Zitnan R, Schönhusen U, Pfannkuche H, Hudakova M, Metges CC (2014) Diet effects on glucose absorption in the small intestine of neonatal calves: importance of intestinal mucosal growth, lactase activity, and glucose transporters. J Dairy Sci. https://doi.org/10.3168/jds.2014-8391 CrossRefGoogle Scholar
- Tyzzer EE (1912) Cryptosporidium parvum (sp. nov.), a coccidium found in the small intestine of the common mouse. Arch Protistenkd 26:394–412Google Scholar