Characterisation of Proton Pump Antibodies and Stomach Pathology in Gastritis Induced by Neonatal Immunisation without Adjuvant

Abstract

It has previously been reported that neonatal BALB.D2 mice injected with native proton pump antigens without adjuvant develop an irreversible gastritis (Claeys et al, 1997). The ease of inititating gastritis in the neonate stands in contrast with the difficulty in initiating gastritis in adult mice that require repeated immunisation in adjuvant that is reversible following cessation of immunisation (Scarff et al, 1997). In view of these contrasting observations, we set out to ascertain whether we could confirm the observations in neonatal mice as well as further characterise the pathology and the autoantibody response. We found that neonatal gastritis-susceptible BALB/c mice (n=12), immunised with either pig or mouse gastric membranes in the absence of adjuvant, develop gastritis without circulating antibody to parietal cells detected by immunofluorescence, a hallmark of murine and human gastritis (Toh et al, 1997). However, mice immunized with pig gastric membranes (n=6) had circulating antibodies reactive by immunofluorescence to recombinant α and/or β subunit of gastric H⁺/K⁺-ATPase expressed by insect cells (Sfα and Sfβ). Four mice from this cohort with antibodies to Sfβ also had reactivity to gastric H⁺/K⁺-ATPase by ELISA, and 3 immunoblotted the β but not the a subunit of the ATPase. In the cohort of mice immunised with mouse gastric membranes (n=6), four produced antibodies reactive by immunofluorescence to Sfα, two of which were also reactive to Sfβ and one developed antibodies detected by ELISA to gastric H⁺/K⁺-ATPase. However, no members of this cohort had antibodies reactive by immunoblot-ting to either the p or a subunit of the ATPase. In all cases gastritic stomachs were characterised by areas deficient in ribosome-rich zymogenic cells and marked reductions in H⁺/K⁺-ATPase-positive parietal cells. Metaplasia detected by Maxwell stain, as clusters of mucus-producing cells throughout gastric units, were non-reactive to stomach mucin autoantibody suggesting the mucins comprise other and/or aberrant form(s). Compared to our previous observations in adult mice, our present data confirms that gastric autoimmunity is more readily induced in the neonate than the adult. Our data also affirms that while the neonatal immune system can mount a damaging inflammatory cellular immune response to gastric antigens, it develops an altered antibody response.

Key Words:

• immunisation
• experimental murine gastritis
• H⁺/K⁺ ATPase