Immunochemical detection of human lactoferrin in feces as a new marker for inflammatory gastrointestinal disorders and colon cancer

Uchida, Kazuo; Matsuse, Ryoichi; Tomita, Shinobu; Sugi, Kazunori; Saitoh, Osamu; Ohshiba, Saburo

doi:10.1016/0009-9120(94)90027-2

Cited by 74 publications

(47 citation statements)

References 12 publications

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“…On mucosal surfaces, iron is mainly sequestered by Lf, and its concentration has been used as a parameter to identify intestinal inflammatory disorders, such as ulcerative colitis (Kane et al, 2003;Masson et al, 1966;Uchida et al, 1994). This study was undertaken to investigate whether E. histolytica is able to use Lf as a source of iron for growth.…”

Section: Discussionmentioning

confidence: 99%

Human hololactoferrin: endocytosis and use as an iron source by the parasite Entamoeba histolytica

et al. 2005

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Entamoeba histolytica is an enteric protozoan that exclusively infects human beings. This parasite requires iron for its metabolic functions. Lactoferrin is a mammalian glycoprotein that chelates extracellular iron on mucosal surfaces, including the surface of the large intestine, where E. histolytica initiates infection. This work examined the interaction in vitro of E. histolytica trophozoites with human hololactoferrin (iron-saturated lactoferrin). A minimum concentration of 50 mM Fe from hololactoferrin supported growth of the amoeba. Amoebic binding sites for hololactoferrin were different from those for human apolactoferrin, holotransferrin and haemoglobin. One amoebic hololactoferrrin-binding polypeptide of 90 kDa was found, which was not observed after treatment of trophozoites with trypsin. Hololactoferrin-binding-protein levels increased in amoebas starved of iron, or grown in hololactoferrin. Internalization of hololactoferrin was inhibited by filipin. Endocytosed hololactoferrin colocalized with an anti-chick embryo caveolin mAb in amoebic vesicles, and lactoferrin was further detected in acidic vesicles; amoebic caveolin of 22 kDa was detected by Western blotting using this antibody. Cysteine proteases from amoebic extracts were able to cleave hololactoferrin. Together, these data indicate that E. histolytica trophozoites bind to hololactoferrin through specific membrane lactoferrin-binding proteins. This ferric protein might be internalized via caveolae-like microdomains, then used as an iron source, and degraded.

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Section: Discussionmentioning

confidence: 99%

Human hololactoferrin: endocytosis and use as an iron source by the parasite Entamoeba histolytica

et al. 2005

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“…Thus, lactoferrin appears to be particularly important in the health and function of the intestinal tract. It greatly reduces inflammation in some bowel diseases, 36 and lactoferrin concentration has been found to be augmented in the large intestine in patients with colitis or active ulcerative colitis, colon cancer, 37 colon disorders and in other pathologies. 38 These results indicate the crucial participation of lactoferrin at the large intestine level.…”

Section: Amoebicidal Effect Of Human Milk Is Due To the Protein Fractmentioning

confidence: 99%

Amoebicidal Activity of Milk, Apo-lactoferrin, sIgA and Lysozyme

León-Sicairos¹,

López-Soto²,

Reyes-López³

et al. 2006

Clinical Medicine & Research

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Entamoeba histolytica is a pathogenic, parasitic protozoan that can infect the large intestine and cause amoebiasis, an illness responsible for at least 100,000 deaths annually worldwide. The capacity of human milk to kill amoebas 6,15 and the amoebicidal action of secretory immunoglobulin type A (sIgA) 16 have been reported. However, it is unknown which other components of milk are amoebicides. As milk contains several microbicidal components, we fractionated milk and tested each fraction against trophozoites in axenic cultures to describe any amoebicidal effects of other components of human and bovine milk such as lactoferrin, lysozyme and sIgA. We also sought to identify any combined effect among these molecules. Objectives: To identify amoebicidal components in human milk and the effect of iron on the amoebicidal activity. Original ResearchDesign: Investigation in axenic cultures of Entamoeba histolytica trophozoites.Methods: Amoebas were treated with 5%-20% of human, bovine and swine milk, with 10% of human milk fractions (i.e., casein, proteins except casein and fat) or with 1 mg/ml of human milk apo-lactoferrin, human secretory immunoglobulin type A (sIgA) and chicken egg-white lysozyme (i.e., purified proteins). Milk proteins were detected using immunoblot. Confocal microscopy was used to define the interaction of milk proteins (100 µM each) and amoebas. Experiments were done at least three times in triplicate, and mean and standard deviations were calculated.Results: Human and bovine milk were amoebicidal showing a concentration-dependent effect. The amoebicidal effect was increased in the absence of iron. Milk protein fractions, with the exception of casein, were the components responsible for the amoebicidal activity found. Apo-lactoferrin, sIgA and lysozyme were identified in the amoebicidal milk protein fraction. Apo-lactoferrin showed the major amoebicidal effect. These proteins, either alone or in combination, showed a killing effect on the trophozoites.They bound to the amoebic membrane causing cell rounding, lipid disruption and damage. Conclusions:Milk proteins such as apo-lactoferrin, sIgA and lysozyme are able to kill Entamoeba histolytica trophozoites.This study confirms the importance of feeding breast milk to newborns.

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“…An alternative approach to the assessment of the presence of intestinal inflammation [12] is to analyze the whole gut lavage fluid or to quantitate the protease resistant neutrophil derived proteins such as lactoferrin [13,14] in the patient's feces, and this approach can be non-invasive for the patient. Calprotectin is one of these proteins.…”

Section: Introductionmentioning

confidence: 99%

Clinical value of fecal calprotectin in determining disease activity of ulcerative colitis

Xiang¹,

Ouyang²,

Li³

et al. 2008

WJG

101

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AIM:To investigate possibility and clinical application of fecal calprotectin in determining disease activity of ulcerative colitis (UC). METHODS:The enzyme-linked immunosorbent assay (ELISA) was used to measure the concentrations of calprotectin in feces obtained from 66 patients with UC and 20 controls. C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), acid glycoprotein (AGP) were also measured and were compared with calprotectin in determining disease activity of UC. The disease activity of UC was also determined by the Sutherland criteria. RESULTS:The fecal calprotectin concentration in the patients with active UC was signifi cantly higher than that in the inactive UC and in the controls (402.16 ± 48.0 μg/g vs 35.93 ± 3.39 μg/g, 11.5 ± 3.42 μg/g, P < 0.01).The fecal calprotectin concentration in the inactive UC group was signifi cantly higher than that in the control group (P < 0.05). A signifi cant difference was also found in the patients with active UC of mild, moderate and severe degrees. The area under the curve of the receiver operating characteristics (AUC ROC ) was 0.975, 0.740, 0.692 and 0.737 for fecal calprotectin, CRP, ESR and AGP, respectively. There was a strong correlation between the fecal calprotectin concentration and the endoscopic gradings for UC (r = 0.866, P < 0.001). CONCLUSION:Calprotectin in the patient's feces can refl ect the disease activity of UC and can be used as a rational fecal marker for intestinal infl ammation in clinical practice. This kind of marker is relatively precise, simple and noninvasive when compared with other commonlyused markers such as CRP, ESR and AGP.

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Immunochemical detection of human lactoferrin in feces as a new marker for inflammatory gastrointestinal disorders and colon cancer

Cited by 74 publications

References 12 publications

Human hololactoferrin: endocytosis and use as an iron source by the parasite Entamoeba histolytica

Human hololactoferrin: endocytosis and use as an iron source by the parasite Entamoeba histolytica

Amoebicidal Activity of Milk, Apo-lactoferrin, sIgA and Lysozyme

Clinical value of fecal calprotectin in determining disease activity of ulcerative colitis

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