A method of obtaining, processing, and analyzing human intestinal secretions for antibody content.
Human intestinal secretions can be readily obtained using a commercially available intestinal lavage solution. Although such secretions contained abundant protease activity, significant loss of immunoglobulins was prevented by the addition of a mixture of protease inhibitors. The total content of IgA, IgM, and IgG antibody in secretions was measured using sandwich ELISA. In the secretions of ten normal volunteers IgA was most abundant (197 micrograms/ml +/- 103 SD) followed by IgM (12.5 micrograms/ml +/- 6.8 SD) and IgG (0.24 micrograms/ml +/- 0.04 SD). The IgA in secretions was predominantly secretory IgA as shown by sucrose density centrifugation. The effect of intestinal secretions on the sensitivity of the antigen-specific ELISA was tested by adding murine myeloma IgA anti-TNP added to samples of human secretions. IgA anti-TNP activity could be detected as low as 1 ng/ml, and there was no evidence of interference with the ELISA by other constituents in the secretions. Using these methods an antigen-specific secretory IgA anti-cholera toxin B subunit response in the secretions of volunteers given an oral B subunit vaccine was readily demonstrated.
Gaspari
M M
MM
Department of Medicine, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298.
Brennan
P T
PT
Solomon
S M
SM
Elson
C O
CO
D000906:Antibodies / Q000032:analysis*; D000907:Antibodies, Bacterial / Q000032:analysis; D002772:Cholera Toxin / Q000276:immunology; D004573:Electrolytes; D004797:Enzyme-Linked Immunosorbent Assay; D005243:Feces / Q000276:immunology*; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis*; D010446:Peptide Fragments / Q000276:immunology; D011092:Polyethylene Glycols; D011480:Protease Inhibitors; D013048:Specimen Handling / Q000379:methods*; D014734:Vibrio cholerae / Q000276:immunology
D000906:Antibodies; D000907:Antibodies, Bacterial; D004573:Electrolytes; C033608:Golytely; D007136:Immunoglobulins; D010446:Peptide Fragments; D011480:Protease Inhibitors; D011092:Polyethylene Glycols; C050310:cholera toxin, B subunit (50-64); D002772:Cholera Toxin