Abstract
Millions of people worldwide are infected with filarial nematodes, responsible for lymphatic filariasis (LF) and other diseases causing chronic disablement. Elimination programs have resulted in a substantial reduction of the rate of infection in certain areas creating a need for improved diagnostic tools to establish robust population surveillance and avoid LF resurgence. Glycans from parasitic helminths are emerging as potential antigens for use in diagnostic assays. However, despite its crucial role in host–parasite interactions, filarial glycosylation is still largely, structurally, and functionally uncharacterized. Therefore, we investigated the glycan repertoire of the filarial nematode Brugia malayi. Glycosphingolipid and N-linked glycans were extracted from several life-stages using enzymatic release and characterized using a combination of MALDI-TOF-MS and glycan sequencing techniques. Next, glycans were purified by HPLC and printed onto microarrays to assess the host anti-glycan antibody response. Comprehensive glycomic analysis of B. malayi revealed the presence of several putative antigenic motifs such as phosphorylcholine and terminal glucuronic acid. Glycan microarray screening showed a recognition of most B. malayi glycans by immunoglobulins from rhesus macaques at different time points after infection, which permitted the characterization of the dynamics of anti-glycan immunoglobulin G and M during the establishment of brugian filariasis. A significant level of IgG binding to the parasite glycans was also detected in infected human plasma, while IgG binding to glycans decreased after anthelmintic treatment. Altogether, our work identifies B. malayi glycan antigens and reveals antibody responses from the host that could be exploited as potential markers for LF.
Highlights
Parasitic worms responsible for several neglected tropical diseases (NTDs) represent a global public health burden and cause disease in hundreds of millions of impoverished people[1,2,3,4,5]
We analyzed N-glycans and GSL glycans isolated from B. malayi adult males, adult females, MF
Up to three of these GlcNAc residues are directly attached to the trimannosyl core possibly extended by additional β(1-4/6) linked GlcNAc elements resulting in N-glycan structures with a maximum of 3 antennae, as described for C. elegans[73]
Summary
Parasitic worms responsible for several neglected tropical diseases (NTDs) represent a global public health burden and cause disease in hundreds of millions of impoverished people[1,2,3,4,5]. N-linked and GSL glycans from approximately 600 adult female worms were extracted, released, labeled as described in the Structural Analysis section above, and pooled in order to obtain a sufficient amount of glycan for microarray generation. Set #2 consisted of 5 individuals obtained from a different study[56,57] in which patients were treated with diethylcarbamazine citrate (DEC) anthelminthic after initial sampling in 1990 Another blood sampling was performed almost 2 years later to address the infection status of the subjects. Journal Pre-proof data are summarized in Table S2.B. Glycan microarray incubation and data analysis Generated B. malayi glycan microarrays were first incubated with the mAbs for validation before human plasma and rhesus macaque sera were screened for antibody binding to glycans. This package, optimized for statistical analysis of microarray data uses a linear model approach to analyze microarray experiments and empirical Bayesian methods to assess significant differences between samples[62,63]
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