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Biomedical subjects

D Magne

Publications and source records attributed to D Magne.

29 records · Page 2Linked to original sources

Cryptosporidiosis-induced impairment of ion transport and Na+-glucose absorption in adult immunocompromised mice.

Electrolyte transport was investigated during chronic cryptosporidiosis in adult anti-interferon-gamma-treated SCID mice by means of Ussing chamber techniques. In basal conditions, infection of immunocompromised mice with Cryptosporidium parvum resulted in a 30% reduction (P < .05) in the ileal short-circuit (Isc) current related to a 28% reduction (P < .05) in tissue conductance compared with controls. The rises in Isc and transepithelial potential difference induced by glucose (10 mM) were significantly reduced by Cryptosporidium infection (P < .01) compared with controls. In contrast, responses to mucosal glutamine were marginally affected. Electrical parameters of the ileum were not affected by the addition of indomethacin or furosemide, in either control or Cryptosporidium-infected mice. Thus, long-term cryptosporidiosis in immunocompromised animals leads to a reduction in net ion exchanges, decreased paracellular shunting, and impaired Na+-glucose cotransport in the ileum, without prostanoid- or enterotoxin-mediated electrogenic Cl- secretion.

Animals↗

Kinetics of mucosal ileal gamma-interferon response during cryptosporidiosis in immunocompetent neonatal mice.

The kinetics of serum and ileal interferon-gamma (IFN-gamma) content were determined during recovery from cryptosporidiosis in NMRI suckling mice. A total of 60 mice aged 4 days were inoculated by intragastric gavage with 10(4) cryptosporidia (n = 30) or phosphate-buffered saline (n = 30). Six animals per group were killed on days 0, 3, 6, 9 and 13 postinoculation. Blood samples and ileum were collected. Experimental infection was followed by a rise in parasite load in the ileum starting on day 3 postinfection, which peaked at day 6 postinoculation. Ileal IFN-gamma levels increased rapidly in parasitized mice from day 3 to day 6, then fell rapidly. These levels were significantly higher than the control values (day 3 P < 0.05, days 6 and 9 P < 0.001). IFN-gamma secretion began before parasite excretion, but the curves of these two parameters correlated positively. Recovery from cryptosporidiosis in immunocompetent neonatal mice is thus associated with an early and marked increase in ileal IFN-gamma content.

Animals↗

Inefficacy of intestinal secretory immune response to Cryptosporidium in acquired immunodeficiency syndrome.

BACKGROUND/AIMS: An alteration of the secretory immune response has been forwarded to explain frequent and chronic mucosal infections in patients with acquired immunodeficiency syndrome (AIDS). The aim of this study was to explore the intestinal immunoglobulin (Ig) secretions in patients with AIDS and their relationships to cryptosporidiosis. METHODS: Patients with AIDS and enteric cryptosporidiosis (n = 12), other enteric infections (n = 10), and no identifiable enteric pathogen (n = 10) and human immunodeficiency virus-seronegative controls (n = 18) were studied. The number of intestinal IgA and IgM plasma cells of the duodenal lamina propria mucosa and total and anti-Cryptosporidium IgA, IgM, and IgG were measured in serum and feces. RESULTS: Although not significantly increased, the number of IgA and IgM plasma cells was greater in patients with AIDS (n = 20) than in controls (n = 5). In feces, total IgA outputs and specific anti-Cryptosporidium IgA levels were significantly higher in patients with AIDS and cryptosporidiosis than in the two other groups of patients with AIDS (P < 0.05 and P < 0.01, respectively) and controls (P < 0.001 and P < 0.01, respectively). Total fecal IgM output and specific anti-Cryptosporidium IgM coproantibodies were increased only in the Cryptosporidium-infected patients relative to the controls (P < 0.05). CONCLUSIONS: Despite the development of pathogen-specific mucosal antibody responses, patients with AIDS and cryptosporidiosis fail to clear the parasite.

Acquired Immunodeficiency Syndrome↗

Intestinal lipid metabolism in suckling rats infected with Giardia duodenalis.

We carried out a quantitative and qualitative analysis of intestinal digestion of neutral lipids in suckling rats infected with Giardia duodenalis. Total lipids were measured after extraction from the contents of the stomach, proximal and distal small bowel, caecum and colon. Amounts gradually fell from the stomach to the colon and were identical in infected animals and controls, although high values were occasionally found in the caecum of infected rats. Relative glyceride quantities were determined by means of high-performance thin-layer chromatography. Triglycerides were absent from the distal small bowel, and only free fatty acids and cholesterol were present in the caecum, reflecting normal digestion of neutral lipids in infected suckling rats. Our results suggest that G. duodenalis does not impair intestinal fat digestion in suckling rats.

Animals↗

Evaluation of intestinal clearance and faecal excretion of alpha 1-antiproteinase and immunoglobulins during Crohn's disease and ulcerative colitis.

The intestinal clearance of alpha 1-antiproteinase, monomeric IgA and IgG, and the daily fecal output of polymeric IgA and IgM were investigated in patients with inflammatory bowel diseases (inactive and active Crohn's disease, ulcerative colitis) and in a control group. The intestinal clearance of alpha 1-antiproteinase was significantly increased in all patients with inflammatory bowel diseases (p less than 0.01), irrespective of the grade of the disease. In contrast, increases in intestinal clearances of monomeric IgA and IgG were more closely related to the severity of the intestinal lesions. The associate determination of these three quantities should therefore be of interest for monitoring the degree of intestinal bowel inflammation. Faecal output of polymeric IgA was significantly increased in active intestinal disease (p less than 0.01), whereas faecal IgM levels were not. The determination of the faecal output of polymeric IgA should contribute to the assessment of the activity of inflammatory bowel diseases, and may provide insight into the activation of the mucosal immune system.

Adolescent↗

Factors influencing the reaction of alpha 1-fetoprotein with concanavalin A and Lens culinaris agglutinin in crossed affinoimmunoelectrophoresis.

Concanavalin A (Con A) and lentil lectin (LCA) analysis of alpha-fetoprotein (AFP) glycosylation heterogeneity is used in a variety of clinical situations. We studied the influence of analytical conditions on the separation of AFP glycoforms by using lectin-crossed affinoimmunoelectrophoresis, regardless of the AFP concentration, which can vary over a wide range in biological fluids. We defined the optimal concentration of Con A (2 g/L) and LCA (0.35 g/L) in the first-dimension gel, together with the optimum antigen (AFP)/antibody ratio in the second-dimension gel. The presence of protein in the diluent used for AFP samples was found to change the shape of crossed affinoimmunoelectrophoresis patterns without changing the percentage composition of AFP fractions. The within-run CV was less than 4% for both lectins, and the between-run CV was less than 6.3%. The minimal quantity of AFP that provided a visible pattern with both lectins was 4 ng, corresponding to 50 microL of an 80 micrograms/L AFP sample. These technical conditions allow the cellular origin of AFP to be determined, regardless of the concentration in the sample. Typical AFP lectin patterns of secreting tumors are compared with fetal and cord serum AFP.

Amniotic Fluid↗

Role of cytoskeleton and surface lectins in Giardia duodenalis attachment to Caco2 cells.

An in vitro model of Giardia duodenalis and the Caco2 cell line enable the study of parameters that could play a part in trophozoite attachment. We explored the role of membranous lectins of G. duodenalis in attachment-inhibition studies using carbohydrates in solution. Attachment rates were reduced by 14% and 23% in the presence of 100 mmol/l mannose-6-phosphate and glucose, respectively, as compared with control values. No significant modification was observed after trophozoite trypsinization at room temperature or at 37 degrees C. The inhibitory effects of colchicine (35%) and nocodazole (70%) suggest a primordial role of the cytoskeleton; microtubules appear to be the principal effectors of trophozoite fixation. Scanning electron microscopy revealed circular imprints on the Caco2 brush border after trophozoite detachment. The mechanisms of attachment of G. duodenalis to intestinal enterocyte-like cells in culture are thus essentially of the mechanical or hydrodynamic type; surface lectins would appear to intervene in the specificity for duodenal cells.

Animals↗

Adherence and multiplication of Giardia intestinalis on human enterocyte-like differentiated cells in vitro.

This report describes a technique for studying the adherence and growth of Giardia intestinalis trophozoites (strains PARIS/86/LCF/1, PARIS/86/LCF/2 and PARIS/88/LCF/8) using the human colon carcinoma cell line Caco2. Giardia trophozoites were cultured with Caco2 cells in a modified HSP3 culture medium. The biochemical differentiation of Caco2 cells was established by an increase in sucrase isomaltase activities to values of 4.51 +/- 0.90 and 10.39 +/- 3.00 milliunits/mg protein for 8- and 12-day-old cultures, respectively. Giardia, adherence to 8- and 12-day-old Caco2 cells reached a value of greater than 75% after 60 and 30 min, respectively. Adherence diminished significantly at 24 degrees C and was almost undetectable at 4 degrees C. Depletion of divalent cations reduced the proportion of adherent trophozoites by up to 46%. Adherence was pH-independent between pH 6.0 and 7.6. Parasite growth increased when Caco2 cell monolayers were used instead of axenic cultures. This in vitro human cell model may contribute to the study of the mechanisms and factors involved in the host-parasite interaction.

Animals↗

Determination of anti-Cryptosporidium coproantibodies by time-resolved immunofluorometric assay.

The role of the mucosal immune response against Cryptosporidium has been suggested by studies on the therapeutic effects of hyperimmune colostrum. In order to study the intestinal response to this infection, we have developed a sandwich-type time-resolved immunofluorometric assay for the determination of anti-Cryptosporidium coproantibodies. This assay has the inherent sensitivity of an immunoassay without the problems due to background responses from other biological compounds, and is thus suitable for faecal samples. The intra-assay coefficients of variation (5.1%, 4.6%, and 5.8% for immunoglobulins (Ig) A, M and G respectively), inter-assay coefficients of variation (9.4%; 10.5% and 12.2% for IgA, IgM and IgG, respectively) and specificity (100% for all 3 isotypes) were all satisfactory. Using this assay to study 12 patients with the acquired immune deficiency syndrome (AIDS) who were infected with cryptosporidiosis, we found a marked elevation of anti-Cryptosporidium IgA and IgM coproantibody titres relative to 18 healthy control values, but no correlation with the gravity of the infection in terms of oocyst shedding. These results suggest that a non-protective mucosal immune response develops to Cryptosporidium in AIDS patients.

AIDS-Related Opportunistic Infections↗