Effect of primary immunization on pulmonary clearance of nontypable Haemophilus influenzae

Abstract

Nontypable Haemophilus influenzae (NTHI) is being increasingly recognized as a cause of both adult pneumonia and acute infectious exacerbations in chronic bronchitis. We used a mouse model to study the immune enhancement of pulmonary clearance of NTHI after a primary immunization. BALB/c mice were immunized with whole NTHI either by intraperitoneal (i.p.) or intratracheal (i.t.) routes. There was 10-fold more NTHI-directed antibody detected in the serum of the i.p.-immunized mice than in the serum from the i.t.-immunized animals. Western blot analysis revealed that these antibodies were directed against both NTHI lipooligosaccharide and the various outer membrane proteins of NTHI. The development of NTHI-directed antibodies in serum was associated with significant enhancement of early pulmonary clearance of NTHI. Six hours after delivery of an endobronchial challenge with NTHI, the i.p.-immunized mice had cleared most of the organisms from their lungs, while the i.t.-immunized mice did not clear NTHI any more rapidly than did unimmunized mice. Serum from the i.p.-immunized mice caused more than 99% uptake of NTHI in an in vitro opsonophagocytic assay, while serum from i.t.-immunized mice stimulated little or no phagocytosis of this organism. Opsonophagocytosis of NTHI was obtained with bronchoalveolar lavage (BAL) fluid collected from i.p.-immunized mice 6 h after, but not before, an endobronchial challenge with NTHI. Intravenous injection of an opsonic IgG monoclonal antibody directed against NTHI lipooligosaccharide resulted in both the appearance of this antibody in the alveolar spaces of the unperturbed lung and enhanced pulmonary clearance of NTHI. These data indicate that the i.p. (systemic) route of immunization is more effective than the i.t. route in establishing pulmonary immunity to NTHI in this model system. Furthermore, immune enhancement of clearance of NTHI from the lungs after a primary immunization apparently results from the exudation of opsonic and bactericidal antibodies from the serum into the alveolae in response to the inflammatory challenge.