Purification and functional characterization of mucosal IgA from vaccinated and SIV-infected rhesus macaques

- Mucosal IgA was affinity purified from rhesus macaque fecal homogenates.
- Large quantities of fecal IgA could be purified enabling multiple functional assays.
- Purified fecal IgA mediated neutralization, phagocytosis, and transcytosis inhibition.
- Mucosal IgA from SIV-vaccinated vs. infected macaques exhibited functional differences.
- Evaluation of fecal mucosal IgA is potentially applicable to human clinical studies.

Vaccine-induced mucosal antibodies are often evaluated using small volumes of secretory fluids. However, fecal matter containing mucosal IgA is abundant. We purified fecal IgA from five SIV-vaccinated and five SIV-infected rhesus macaques by sequential affinity chromatography. The purified IgA was dimeric by native PAGE, contained secretory component, and was analogous to IgA in colostrum and vaginal fluid by western blot. IgA from one infected and four vaccinated animals neutralized H9-derived SIVmac251 with IC50s as low as 1 μg/mL. Purified IgAs inhibited transcytosis and exhibited phagocytic activity, the latter significantly correlated with SIVmac251 Env-specific IgA in the purified samples. Among different affinity resins, peptide M was optimal compared to jacalin, anti-monkey IgA and SSL7 for IgA purification, as confirmed using tandem peptide M/anti-monkey IgA columns. Fecal IgA provided material sufficient for several assays relevant to protective efficacy, and was shown to be multifunctional. Our approach is potentially applicable to human clinical studies.