Effect of secretory IgA on transepithelial passage of bacteria across the intact ileum in vitro.
BACKGROUND
Bacterial translocation is a process believed to result in nosocomial infections. Secretory IgA (sIgA) may have a role in the prevention of translocation by its ability to bind and aggregate bacteria, a function termed "immune exclusion." The present study was done to determine the effect of specific binding of sIgA to bacteria on the movement of these organisms across the intact epithelial membrane.


METHODS
Bacterial translocation across intact intestinal segments of rats were assessed in vitro using the Ussing model. Secretory IgA (0.25 mg per mL) from pooled human colostrum was added to the perfused segments of ileum in the Ussing system. Subsequently, the membranes were exposed to 5 x 10(9) cfu per mL Escherichia coli on their mucosal side. A second experiment tested the effect of human IgG when perfused with E. coli using the same preparation. All experiments had paired matched rats in a control group without immunoglobulin. The ability of sIgA and IgG to bind to E. coli was studied by an in vitro assay, as well as by transmission electron microscopy and immunofluorescence of random IgA/E. coli experiments. Measurements obtained in all experimental and control groups were the incidence and amount of bacterial passage and the potential difference generated by the intestinal segments (an index of viability).


RESULTS
There were no differences in potential difference between control and experimental groups in either of the two experiments. Secretory IgA bound E. coli and completely prevented passage of E. coli as compared with rats in the control group. IgG bound E. coli; however, the incidence of passage was equal to that of rats in the control group. However, the presence of IgG resulted in a significantly reduced number of bacteria that passed when compared with controls (p < 0.05). Electron microscopic studies revealed intact surface morphology and immunofluorescence revealed aggregates of IgA and E. coli on the mucosal, but not submucosal, surface of the ileal membranes.


CONCLUSIONS
This study provides direct evidence of immune exclusion by sIgA. When bound to bacteria, it prevents passage across a morphologically intact segment of viable intestinal tissue.
Department of Pediatric Surgery, Children's Hospital of Pittsburgh, PA.
C T
CT
Albanese
S D
SD
Smith
S
S
Watkins
A
A
Kurkchubasche
R L
RL
Simmons
M I
MI
Rowe
D000818:Animals; D004926:Escherichia coli / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D007082:Ileum / Q000166:cytology / Q000276:immunology / Q000382:microbiology; D007071:Immunoglobulin A, Secretory / Q000502:physiology*; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000276:immunology* / Q000382:microbiology*; D008297:Male; D008854:Microscopy, Electron; D051381:Rats; D017207:Rats, Sprague-Dawley
D007071:Immunoglobulin A, Secretory