Effects of sample processing on the measurement of specific intestinal IgA immune responses

Abstract

The effects of techniques commonly used in the collection and processing of human intestinal fluid on the specific secretory immunoglobulin A (sIgA) response following oral immunization with the live typhoid vaccine Salmonella typhi Ty21a were examined. It was observed that the failure to adjust specific intestinal anti-typhoid lipopolysaccharide IgA antibody titres for total secretory IgA resulted in a false-negative detection rate of 19.8% and a false-positive detection rate of 7.4%. Furthermore, these specific responses were significantly diminished if the intestinal fluid was subjected to heat inactivation to reduce intestinal protease activity (p=0.0083), but were not affected if stored at −70°C for up to 1 year, without heat inactivation. It was concluded that in the processing of the intestinal fluid samples for specific sIgA determination heat inactivation significantly reduced specific sIgA titres, and that the failure to adjust absolute titres for total sIgA content resulted in a significant false-negative detection rate.