{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T07:04:59Z","timestamp":1781247899389,"version":"3.54.1"},"reference-count":49,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2026,6,10]],"date-time":"2026-06-10T00:00:00Z","timestamp":1781049600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>For Entamoeba histolytica to establish an infection, it must employ several mechanisms of pathogenicity to produce and secrete virulence factors that allow the parasite to adhere to and finally colonize and invade the host. However, in the intestinal epithelium, trophozoites (amoebae) encounter lactoferrin (Lf), a glycoprotein of the first line of defense, together with immunoglobulins and other molecules. We previously reported that iron-free bovine Lf (bLf) could kill amoebae both in vitro and in animal models of intestinal and hepatic amoebiasis. In this work, selected pathogenic mechanisms were evaluated in trophozoites by exposing cultures to sublethal concentrations of bLf to determine which amoebic functions could be altered. At a sublethal bLf concentration, this glycoprotein was removed from the trophozoite. In the presence of erythrocytes, bLf colocalized with erythrocytes at the capping site; this was observed by confocal microscopy of living cells. In addition, the erythrophagocytosis rate, proteolytic activity, adhesion, and cytotoxic properties towards Caco2 colonic cancer cells were reduced in the presence of bLf. Lactoferrin could be a particularly important protein that naturally protects colonic epithelial cells from E. histolytica infection.<\/jats:p>","DOI":"10.3390\/ijms27125257","type":"journal-article","created":{"date-parts":[[2026,6,11]],"date-time":"2026-06-11T02:59:31Z","timestamp":1781146771000},"page":"5257","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Low-Iron Bovine Lactoferrin Affects Adhesion, Erythrophagocytosis, Cytotoxicity, and Proteolytic Activity in Entamoeba histolytica Trophozoites"],"prefix":"10.3390","volume":"27","author":[{"given":"Magda","family":"Reyes-L\u00f3pez","sequence":"first","affiliation":[{"name":"Departamento de Biolog\u00eda Celular, Centro de Investigaci\u00f3n y de Estudios Avanzados (Cinvestav-IPN), Ave. IPN 2508, Mexico City 07360, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5620-7284","authenticated-orcid":false,"given":"Christian","family":"\u00c1valos-G\u00f3mez","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Celular, Centro de Investigaci\u00f3n y de Estudios Avanzados (Cinvestav-IPN), Ave. IPN 2508, Mexico City 07360, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2546-2792","authenticated-orcid":false,"given":"Gerardo","family":"Ram\u00edrez-Rico","sequence":"additional","affiliation":[{"name":"Laboratorio de Patolog\u00eda Molecular Veterinaria, Facultad de Estudios Superiores de Cuautitl\u00e1n, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Cuautitl\u00e1n Izcalli 54714, Estado de Mexico, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jes\u00fas","family":"Serrano-Luna","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Celular, Centro de Investigaci\u00f3n y de Estudios Avanzados (Cinvestav-IPN), Ave. IPN 2508, Mexico City 07360, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mireya de la","family":"Garza","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Celular, Centro de Investigaci\u00f3n y de Estudios Avanzados (Cinvestav-IPN), Ave. IPN 2508, Mexico City 07360, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"151358","DOI":"10.1016\/j.ijmm.2019.151358","article-title":"Intestinal amoebiasis: 160 years of its first detection and still remains as a health problem in developing countries","volume":"310","author":"Carrero","year":"2020","journal-title":"Int. J. Med. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"110100","DOI":"10.1016\/j.intimp.2023.110100","article-title":"Entamoeba histolytica induced NETosis and the dual role of NETs in amoebiasis","volume":"118","author":"Roy","year":"2023","journal-title":"Int. Immunopharmacol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"108841","DOI":"10.1016\/j.exppara.2024.108841","article-title":"Role of various virulence factors involved in the pathogenesis of Entamoeba histolytica","volume":"266","author":"Mathur","year":"2024","journal-title":"Exp. Parasitol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Boettner, D.R., Huston, C.D., Linford, A.S., Buss, S.N., Houpt, E., Sherman, N.E., and Petri, W.A. (2008). Entamoeba histolytica Phagocytosis of Human Erythrocytes Involves PATMK, a Member of the Transmembrane Kinase Family. PLoS Pathog., 4.","DOI":"10.1371\/journal.ppat.0040008"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.mib.2015.07.009","article-title":"Taking a bite: Amoebic trogocytosis in Entamoeba histolytica and beyond","volume":"28","author":"Ralston","year":"2015","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1046\/j.1365-2958.1998.00662.x","article-title":"Isolation and molecular characterization of a surface-bound proteinase of Entamoeba histolytica","volume":"27","author":"Jacobs","year":"1998","journal-title":"Mol. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Arg\u00fcello-Garc\u00eda, R., Carrero, J.C., and Ortega-Pierres, M.G. (2023). Extracellular Cysteine Proteases of Key Intestinal Protozoan Pathogens\u2014Factors Linked to Virulence and Pathogenicity. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241612850"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/0166-6851(85)90041-6","article-title":"Subcellular fractionation of amoebapore and plasma membrane components of Entamoeba histolytica using self-generating Percoll gradients","volume":"14","author":"Rosenberg","year":"1985","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"178","DOI":"10.1016\/S0169-4758(97)01038-7","article-title":"Amoebapores","volume":"13","author":"Leippe","year":"1997","journal-title":"Parasitol. Today"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/j.pt.2004.10.009","article-title":"Ancient weapons: The three-dimensional structure of amoebapore A","volume":"21","author":"Leippe","year":"2005","journal-title":"Trends Parasitol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1016\/S0140-6736(03)12830-9","article-title":"Amoebiasis","volume":"361","author":"Stanley","year":"2003","journal-title":"Lancet"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1016\/j.arcmed.2005.10.005","article-title":"Host Tissue Invasion by Entamoeba histolytica Is Powered by Motility and Phagocytosis","volume":"37","year":"2006","journal-title":"Arch. Med. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/S0014-4894(02)00106-6","article-title":"Entamoeba histolytica: Acute granulomatous intestinal lesions in normal and neutrophil-depleted mice","volume":"101","author":"Tsutsumi","year":"2002","journal-title":"Exp. Parasitol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Le\u00f3n-Sicairos, N., Ordaz-Pichardo, C., Carrero, J., and de la Garza, M. (2017). Lactoferrin in the Battle against Intestinal Parasites: A Review. Natural Remedies in the Fight Against Parasites, InTech.","DOI":"10.5772\/66819"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1080\/08927014.2013.773317","article-title":"Mini-review: Lactoferrin: A bioinspired, anti-biofilm therapeutic","volume":"29","author":"Ammons","year":"2013","journal-title":"Biofouling"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Kowalczyk, P., Kaczy\u0144ska, K., Kleczkowska, P., Bukowska-O\u015bko, I., Kramkowski, K., and Sulejczak, D. (2022). The Lactoferrin Phenomenon-A Miracle Molecule. Molecules, 27.","DOI":"10.3390\/molecules27092941"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Rosa, L., Cutone, A., Lepanto, M.S., Paesano, R., and Valenti, P. (2017). Lactoferrin: A Natural Glycoprotein Involved in Iron and Inflammatory Homeostasis. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18091985"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.biochi.2008.05.015","article-title":"Antimicrobial properties of lactoferrin","volume":"91","author":"Jenssen","year":"2009","journal-title":"Biochimie"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Reyes-L\u00f3pez, M., Ram\u00edrez-Rico, G., Serrano-Luna, J., and de la Garza, M. (2022). Activity of Apo-Lactoferrin on Pathogenic Protozoa. Pharmaceutics, 14.","DOI":"10.3390\/pharmaceutics14081702"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4550","DOI":"10.1128\/iai.63.11.4550-4552.1995","article-title":"Giardicidal activity of lactoferrin and N-terminal peptides","volume":"63","author":"Turchany","year":"1995","journal-title":"Infect. Immun."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"18020","DOI":"10.1038\/s41598-018-36563-1","article-title":"Lactoferrin and lactoferricin endocytosis halt Giardia cell growth and prevent infective cyst production","volume":"8","author":"Frontera","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1139\/bcb-2016-0079","article-title":"Parasiticidal effect of synthetic bovine lactoferrin peptides on the enteric parasite Giardia intestinalis","volume":"95","author":"Nazmi","year":"2017","journal-title":"Biochem. Cell Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1111\/j.1550-7408.1997.tb05694.x","article-title":"Ultrastructural effects of lactoferrin binding on Giardia lamblia trophozoites","volume":"44","author":"Turchany","year":"1997","journal-title":"J. Eukaryot. Microbiol."},{"key":"ref_24","first-page":"3821","article-title":"Effect of lactoferrin protein on red blood cells and macrophages: Mechanism of parasite-host interaction","volume":"9","author":"Anand","year":"2015","journal-title":"Drug Des. Dev. Ther."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0014-4894(87)90072-5","article-title":"Plasmodium falciparum: Inhibition in vitro with lactoferrin, desferriferrithiocin, and desferricrocin","volume":"63","author":"Fritsch","year":"1987","journal-title":"Exp. Parasitol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1111\/j.1550-7408.1988.tb04086.x","article-title":"Trypanosoma cruzi: A specific surface marker for the amastigote form","volume":"35","author":"Lima","year":"1988","journal-title":"J. Protozool."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3859","DOI":"10.1099\/mic.0.28121-0","article-title":"Human hololactoferrin: Endocytosis and use as an iron source by the parasite Entamoeba histolytica","volume":"151","author":"Arroyo","year":"2005","journal-title":"Microbiology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"106","DOI":"10.3121\/cmr.4.2.106","article-title":"Amoebicidal activity of milk, apo-lactoferrin, sIgA and lysozyme","volume":"4","year":"2006","journal-title":"Clin. Med. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1139\/o11-084","article-title":"Effect of bovine lactoferrin in a therapeutic hamster model of hepatic amoebiasis","volume":"90","year":"2012","journal-title":"Biochem. Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1139\/o2012-008","article-title":"Oral lactoferrin treatment resolves amoebic intracecal infection in C3H\/HeJ mice","volume":"90","author":"Carrero","year":"2012","journal-title":"Biochem. Cell Biol."},{"key":"ref_31","first-page":"327","article-title":"Microbicidal action of lactoferrin and lactoferricin and their synergistic effect with metronidazole in Entamoeba histolytica","volume":"84","year":"2006","journal-title":"Biochem. Cell Biol."},{"key":"ref_32","first-page":"173","article-title":"Scanning electron microscopy of erythrophagocytosis by Entamoeba histolytica trophozoites","volume":"23","author":"Tsutsumi","year":"1992","journal-title":"Arch. Med. Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1016\/0035-9203(92)90555-Q","article-title":"Entamoeba histolytica: Erythrophagocytosis, collagenolysis, and liver abscess production as virulence markers","volume":"86","author":"Tsutsumi","year":"1992","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Conesa, C., Bell\u00e9s, A., Grasa, L., and S\u00e1nchez, L. (2023). The Role of Lactoferrin in Intestinal Health. Pharmaceutics, 15.","DOI":"10.3390\/pharmaceutics15061569"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Begum, S., Quach, J., and Chadee, K. (2015). Immune Evasion Mechanisms of Entamoeba histolytica: Progression to Disease. Front. Microbiol., 6.","DOI":"10.3389\/fmicb.2015.01394"},{"key":"ref_36","first-page":"626259","article-title":"Erythrophagocytosis in Entamoeba histolytica and Entamoeba dispar: A comparative study","volume":"2014","year":"2014","journal-title":"Biomed. Res. Int."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1137","DOI":"10.1084\/jem.148.5.1137","article-title":"Surface properties of Entamoeba: Increased rates of human erythrocyte phagocytosis in pathogenic strains","volume":"148","author":"Trissl","year":"1978","journal-title":"J. Exp. Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1007\/s00436-012-2820-2","article-title":"Differences in cap formation between invasive Entamoeba histolytica and non-invasive Entamoeba dispar","volume":"111","year":"2012","journal-title":"Parasitol. Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1084\/jem.151.1.184","article-title":"Surface redistribution and release of antibody-induced caps in entamoebae","volume":"151","author":"Acosta","year":"1980","journal-title":"J. Exp. Med."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1006\/expr.1994.1104","article-title":"Entamoeba histolytica: Mechanism of surface receptor capping","volume":"79","year":"1994","journal-title":"Exp. Parasitol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e1283386","DOI":"10.1080\/21688370.2017.1283386","article-title":"Entamoeba histolytica: Host parasite interactions at the colonic epithelium","volume":"5","author":"Cornick","year":"2017","journal-title":"Tissue Barriers"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.micpath.2011.10.005","article-title":"Anoikis potential of Entameba histolytica secretory cysteine proteases: Evidence of contact independent host cell death","volume":"52","author":"Kumar","year":"2012","journal-title":"Microb. Pathog."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1473","DOI":"10.1038\/sj.cdd.4401723","article-title":"Anoikis","volume":"12","author":"Gilmore","year":"2005","journal-title":"Cell Death Differ."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1006\/expr.1998.4258","article-title":"Entamoeba histolytica HM1:IMSS: Hemoglobin-degrading neutral cysteine proteases","volume":"89","author":"Negrete","year":"1998","journal-title":"Exp. Parasitol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1007\/978-1-4939-6352-2_3","article-title":"Production and Purification of Polyclonal Antibodies","volume":"1474","author":"Nakazawa","year":"2016","journal-title":"Methods Mol. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1677","DOI":"10.1126\/science.142.3600.1677","article-title":"Antiserums Prepared with Acrylamide Gel Used as Adjuvant","volume":"142","author":"Weintraub","year":"1963","journal-title":"Science"},{"key":"ref_47","unstructured":"Ordaz-Pichardo, C., Le\u00f3n-Sicairos, N., Canizalez-Roman, A., Cornejo-Cort\u00e9s, M., Ort\u00edz-Estrada, G., and De la Garza, M. (2013). Chapter 7. Lactoferrin: A protein of the innate immune system capable of killing parasitic protozoa. A review. Parasites: Ecology, Diseases and Management, Nova Science Publishers Inc."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"BSR20180850","DOI":"10.1042\/BSR20180850","article-title":"Synthetic bovine lactoferrin peptide Lfampin kills Entamoeba histolytica trophozoites by necrosis and resolves amoebic intracecal infection in mice","volume":"39","author":"Bobes","year":"2019","journal-title":"Biosci. Rep."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"4142","DOI":"10.1128\/iai.59.11.4142-4146.1991","article-title":"A new in vitro model of Entamoeba histolytica adhesion, using the human colon carcinoma cell line Caco-2: Scanning electron microscopic study","volume":"59","author":"Rigothier","year":"1991","journal-title":"Infect. Immun."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/12\/5257\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T06:55:05Z","timestamp":1781247305000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/12\/5257"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,10]]},"references-count":49,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["ijms27125257"],"URL":"https:\/\/doi.org\/10.3390\/ijms27125257","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,10]]}}}