Localization of inhaled trimellitic anhydride to lung with a respiratory lymph node antibody secreting cell response

Abstract

Male Sprague-Dawley rats were exposed to trimellitic anhydride (TMA) by inhalation (500 μg/m 3), 4 hours a day, for 1 to 10 days. TMA was localized to lung cells by immunoelectron microscopy. Enzyme-linked immunosorbent assay (ELISA) and Western blot analysis were used in an attempt to localize TMA to lung lavage proteins. The lung-associated lymph node (LALN) B-lymphocyte response was measured by quantitation of immunoglobulin (Ig)G, IgA, and IgM antibody secreting cells specific for TMA rat serum albumin (TMRSA) by use of the enzyme-linked immunospot assay (ELISPOT) method. The IgG, IgM, and IgA antibody response to TMRSA in serum and lavage fluid was quantitated by ELISA. Lung injury was assessed by the number of external lung hemorrhagic foci and lung weight. Immunoelectron microscopy localized TMA to alveolar and bronchial cells on all exposure days. ELISA detected trace amounts of TMA haptenized lavage proteins that could not be detected by Western blot analysis. A marked increase occurred in lung injury from day 7 to 10. The LALN IgG, IgA, and IgM antibody secreting cell response to TMRSA paralleled measures of lung injury. IgG, IgM, and IgA serum and lavage antibody to TMRSA were correlated with lung injury measures. Lavage and serum IgG antibody levels had the highest correlation with lung injury.