title: Detection of rectal antibodies to HIV-1 by a sensitive chemiluminescent western blot immunodetection method.
abstract: Western blot with a time-dependent enhanced chemiluminescence immunodetection method (ECL-WB) was shown to be 100-fold more sensitive than standard commercial colorimetric Western blots (WB) for detecting serum IgG to human immunodeficiency virus type 1 (HIV-1). ECL-WB was then used to test rectal secretions from 15 HIV-1 infected subjects (HIV+) and 7 uninfected subjects (HIV-) to document local IgG, IgA, and secretory component-associated immunoglobulin (SC-Ig) to HIV-1 proteins. Fourteen of 15 HIV+ subjects had rectal IgA to at least 1 HIV-1 protein, most often to gp41 (80%) or p24 (60%) and 14 (93%) had IgG to gp160, gp120, or gp41. Of seven HIV- subjects, none had detectable bands to HIV-1 proteins. SC-Ig to HIV-1 proteins was detected in all five rectal samples tested. However, the antibody profiles differed from those of rectal IgA, suggesting more than one source of rectal IgA to HIV. ECL-WB requires individual optimization and interpretation for each specimen as well as expensive reagents and is, therefore, not currently applicable to screening assays. However, the method offers promise as a sensitive method to characterize low-level immune responses (IgG, IgA, and SC-Ig) to HIV-1 proteins at local sites such as rectal mucosae.
authors.affiliation: Department of Epidemiology, School of Public Health, University of Washington, Seattle.
authors.forename: O A
authors.initials: OA
authors.lastname: Mohamed
authors.forename: R
authors.initials: R
authors.lastname: Ashley
authors.forename: A
authors.initials: A
authors.lastname: Goldstein
authors.forename: J
authors.initials: J
authors.lastname: McElrath
authors.forename: J
authors.initials: J
authors.lastname: Dalessio
authors.forename: L
authors.initials: L
authors.lastname: Corey
mesh_terms: D015153:Blotting, Western; D015483:HIV Antibodies / Q000032:analysis*; D006679:HIV Seropositivity / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006716:Homosexuality; D006801:Humans; D007071:Immunoglobulin A, Secretory / Q000032:analysis; D007074:Immunoglobulin G / Q000032:analysis; D007413:Intestinal Mucosa / Q000276:immunology / Q000378:metabolism; D008163:Luminescent Measurements; D008297:Male; D012007:Rectum / Q000276:immunology* / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D015483:HIV Antibodies; D007071:Immunoglobulin A, Secretory; D007074:Immunoglobulin G