Validation of a gastrointestinal explant system for measurement of mucosal antibody production.
A gastrointestinal explant culture system was developed and compared to the mononuclear cell extraction and enzyme-linked immunospot assay method for measurement of immunoglobulin A (IgA) and IgG antibody-secreting cells (ASCs) in gastric antral and duodenal biopsies of non-Helicobacter pylori-infected volunteers. IgA and IgG were detected in explant supernatants during 6 to 7 days of culture in all subjects. IgA containing secretory component was also detected throughout the culture period, although peak production occurred only in the first 3 days. During 7 days of culture, the cumulative geometric mean IgA levels produced were 2.2 and 8.02 microg/ml/10 mg of antral and duodenal biopsy tissues, respectively, while the cumulative geometric mean IgG levels were 1.54 and 2.92 microg/ml/10 mg of antral and duodenal biopsy tissues, respectively. Cycloheximide treatment resulted in a >90% reduction in both immunoglobulin classes after 6 days of treatment compared to levels in untreated controls. The detection of IgA and IgG ASCs extracted from biopsies on days 1 and 6 of culture confirmed that the antibody detected was derived from mucosal lamina propria. The IgA and IgG ASC responses were positively correlated with antibody concentrations detected in culture supernatants (r = 0.87 and 0.85, respectively). These results validate the potential usefulness of our gastrointestinal explant system for the evaluation of mucosal effector B-cell function.
Losonsky
G A
GA
Center for Vaccine Development, Division of Infectious Diseases and Tropical Pediatrics and Division of Geographic Medicine, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA. glosonsk@umppa1.ab.umd.edu
Fantry
G T
GT
Reymann
M
M
Lim
Y
Y
D000293:Adolescent; D000328:Adult; D000907:Antibodies, Bacterial / Q000096:biosynthesis*; D000917:Antibody Formation / Q000187:drug effects / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D001706:Biopsy; D018929:Cell Culture Techniques / Q000592:standards*; D003513:Cycloheximide / Q000494:pharmacology; D004386:Duodenum / Q000166:cytology / Q000276:immunology / Q000382:microbiology; D005260:Female; D005753:Gastric Mucosa / Q000166:cytology* / Q000276:immunology* / Q000473:pathology; D016481:Helicobacter Infections / Q000276:immunology*; D016480:Helicobacter pylori / Q000276:immunology*; D006801:Humans; D007070:Immunoglobulin A / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000096:biosynthesis; D007700:Kinetics; D008297:Male; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011706:Pyloric Antrum / Q000166:cytology / Q000276:immunology / Q000382:microbiology; D015203:Reproducibility of Results
D000907:Antibodies, Bacterial; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D011500:Protein Synthesis Inhibitors; D003513:Cycloheximide