Cryptococcus neoformans causes meningitis in 6 to 8% of individuals with AIDS. Recently, immunotherapeutic modalities including antibody therapy have been proposed for the treatment of cryptococcal meningitis in AIDS patients. This is a rational approach because existing antifungal agents fail to eradicate the infection in the setting of profound immunosuppression. Both murine and human antibodies elicited by the investigational cryptococcal capsular polysaccharide vaccine glucuronoxylomannan-tetanus toxoid (GXM-TT) have been shown to be biologically functional in different model systems. The human immunoglobulin M (λ) GXM monoclonal antibody (MAb) 2E9 expresses idiotypes that are also found in naturally occurring anti-GXM antibodies and opsonic GXM-TT sera. However, the specificity of human anti-GXM antibodies and their possible role in protection against cryptococcosis are not known. In an effort to discover epitopes that are recognized by human anti-GXM antibodies, we screened a random decapeptide phage display library with the human anti-GXM MAb 2E9. An enzyme-linked immunosorbent assay (ELISA)-based screening method led to the selection of phages with peptide inserts that bound 2E9 and inhibited 2E9-GXM binding. Analysis of the amino acid sequences of these phages revealed an increased frequency of combinations of QTGLD residues. Inhibition ELISAs demonstrated that phages with QTG/TL/D motifs inhibited 2E9-GXM binding better than phages with different motifs. A peptide synthesized from one of the inhibitory phages, peptide 13 (GMDGT QLDRW), inhibited GXM binding to solid-phase 2E9 and 2E9 binding to solid-phase GXM. Peptide 13 also inhibited the GXM binding of GXM-TT immune sera and naturally occurring serum antibodies from human immunodeficiency virus (HIV)-negative, but nut HIV-positive, individuals. Taken together, our data indicate that the peptide epitopes selected by 2E9 mimic GXM epitopes and that peptide 13 may be a mimotope of a GXM epitope that is recognized by human anti-GXM antibodies.
ASJC Scopus subject areas
- Infectious Diseases