Anti-CD۱۹/CD۸ bispecific T cell engager for the potential treatment of B cell malignancies

B cell lymphoma is a malignancy derived from B cells and consists of two subtypes: Hodgkin and non-Hodgkin lymphoma (NHLs). NHL is the most common type (۹۰ %) of lymphoma, which affects children more frequently. In comparison with toxic and traditional chemotherapy, cancer immunotherapy (targeted therapy) is the best approach to primarily focus on cancer cells, even in chemo-resistant tumors. In ۲۰۱۴, blinatumomab, the first FDA-approved Bispecific T-cell engager (BiTE), was approved for treating acute lymphoblastic leukemia. However, the administration of blinatumomab was accompanied by several adverse effects, including activation of regulatory T cells (Tregs) and cytokine storm. The objective of this study was to produce and evaluate a novel anti-CD۸/CD۱۹ BiTE (αCD۸/CD۱۹) with the potency to target CD۸+ T cells directly. In silico studies were utilized for determining physicochemical properties and predicting ۳D model of αCD۸/CD۱۹. Using molecular docking and molecular dynamics simulation techniques, the stability and binding affinity of the recombinant protein were determined. Western blotting and indirect surface staining were used to evaluate the size accuracy and binding potency of the purified protein. Functionality was assessed by CFSE staining for granzyme B production and cytotoxicity. In silico analysis revealed that the construct has a high affinity for the CD۸ receptor. The αCD۸/CD۱۹ protein was produced with a final concentration of ۱.۹۴ mg/l. The presence of αCD۸/CD۱۹ significantly increased CD۸+ T-cell cytotoxicity in comparison to control groups. The maximum functionality was observed on the ۱۰th day with a ۱۰:۱ effector–to–target ratio. The αCD۸/CD۱۹ bound to CD۸+ and CD۱۹+ cell lines and induced significant granzyme B production and cytotoxic activity in the presence of IL-۲ and tumor target cells. The results demonstrated the potential of αCD۸/CD۱۹ as a promising therapeutic agent for targeted cytotoxicity against CD۱۹-positive cancer cells. Further investigations and clinical studies are warranted to validate the efficacy and safety of αCD۸/CD۱۹ in preclinical and clinical settings.

کلیدواژه ها:

Bispecific T-cell Engager ، B cell malignancies ، Blinatumomab ، CD۸+ Tcell Lymphocytes