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

Purpose 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 lectin from Streptococcus suis , to target globotriaosylceramide (Gb3), a glycosphingolipid overly abundant in many cancer types including Burkitt’s lymphoma. SadP was genetically fused to a scFv of an anti-CD3 antibody, resulting in a bispecific T cell engager (BiTE)-like protein referred to as lectibody. Methods The lectibody was designed as a fusion protein by genetically linking the SadP to the scFv UCHT1 anti-CD3 antibody. Protein expression was carried out in Escherichia coli 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. Results The present study demonstrates that the 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% in-vitro 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 ⁺ cancer cells. Conclusion 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 published StxB-scFv UCHT1 lectibody, therefore opening the possibility for its use in an in-vivo model.