Volume 119, Issue 5, Pages (November 2000)

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Volume 119, Issue 5, Pages 1236-1242 (November 2000) Mannose-binding lectin is a component of innate mucosal defense against Cryptosporidium parvum in AIDS Paul Kelly, Dominic L. Jack, Arifa Naeem, Benson Mandanda, Richard C.G. Pollok, Nigel J. Klein, Malcolm W. Turner, Michael J.G. Farthing Gastroenterology Volume 119, Issue 5, Pages 1236-1242 (November 2000) DOI: 10.1053/gast.2000.19573 Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 1 MBL binding to C. parvum sporozoites demonstrated by immunofluorescence. Sporozoites were incubated with 1750 ng/mL MBL and then FITC–anti-MBL. No fluorescence was detected on control sporozoites incubated without MBL. Gastroenterology 2000 119, 1236-1242DOI: (10.1053/gast.2000.19573) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 2 Representative flow-cytometric profiles of C. parvum. Typical size and granularity profiles of (A) sporozoites and (B) oocysts using identical machine settings. Forward scatter (size) is shown on the x-axis and side scatter (granularity) on the y-axis. Fluorescence histograms of (C) sporozoites and (D) oocysts. The open histogram represents organisms after incubation with 1750 ng/mL MBL and then FITC–anti-MBL; the shaded histogram represents organisms incubated with FITC–anti-MBL alone. (E) Sporozoites incubated with 50 ng/mL MBL (open histogram). Fluorescence units are shown on the x-axis and number of events on the y-axis. Gastroenterology 2000 119, 1236-1242DOI: (10.1053/gast.2000.19573) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 3 Concentration dependence of MBL binding to C. parvum sporozoites. Binding of purified MBL at different concentrations was determined by flow cytometry and expressed as median fluorescence intensity. Each point represents the mean ± SEM of 3 experiments. Gastroenterology 2000 119, 1236-1242DOI: (10.1053/gast.2000.19573) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 4 Density dot plots of MBL binding with C4 deposition on sporozoites after the binding of MBL/MASP. (A) Sporozoites incubated with 100 μg/mL of purified C4 and not incubated with MBL. (B and C) Binding of C4 on sporozoites preincubated with 0.2 and 2 μg/mL MBL, respectively. PE-Cy5, phycoerythrin-Cy5 conjugate. Gastroenterology 2000 119, 1236-1242DOI: (10.1053/gast.2000.19573) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 5 Reducing sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis of duodenal fluid aspirates. The upper gel has been stained silver to show total protein, and the lower gel has been blotted and probed with anti-MBL antibodies using an ECL detection system. M, purified MBL; 1–5, different duodenal fluid aspirates. Gastroenterology 2000 119, 1236-1242DOI: (10.1053/gast.2000.19573) Copyright © 2000 American Gastroenterological Association Terms and Conditions