A novel SadP-scFv UCHT1 lectibody activates T cells and mediates lysis of Burkitt's lymphoma cells

Abnormal glycosylation has been long considered a hallmark of cancer progression. Carbohydrate-binding proteins, also known as lectins, offer a unique way to target glycosylation changes in malignant cells. The present study repurposes SadP, a monomeric lectin from <i>Streptococcus suis</i>, to target globotriaosylceramide (Gb3), a glycosphingolipid overly abundant in many cancer types including Burkitt's lymphoma. The lectibody was designed as a fusion protein by linking the SadP to the scFv UCHT1 anti-CD3 antibody resulting in a bispecific T cell engager (BiTE)-like protein referred to as lectibody. Protein expression was carried out in <i>Escherichia coli</i> and the resulting lectibody was purified using affinity and size exclusion chromatography. The lectibody was tested for its specificity in binding Gb3-positive cancer cells by flow cytometry. T-cell-mediated cytotoxicity was measured in a bioluminescence-based cytotoxicity assay, and T-cell activation was assessed by evaluating CD69 and CD71 expression on PBMCs, incubated with target cells and the lectibody. The present study demonstrates that the monomeric and monovalent SadP-scFv UCHT1 lectibody can redirect T cell cytotoxicity towards Gb3⁺ Burkitt's lymphoma cells, resulting in a dose-dependent target cell lysis up to 65% <i>in vitro</i> at a concentration of 10 nM. In the same experimental setting, negative control cells characterized by a low or absent Gb3 content remained unaffected. Lectibody-induced T cell activation resulted in a significant increase in CD69 and CD71 surface expression in PBMCs incubated with SadP-scFv UCHT1 and Gb3 positive cancer cells. This study highlights the potential of lectins in immunotherapy for the treatment and eradication of malignant cells. The SadP-based lectibody demonstrates improved efficacy and yield when compared to the previously engineered StxB-scFv UCHT1 lectibody, therefore opening the possibility for its use in an <i>in vivo</i> model.