Preparation and In Vitro Evaluation of RITUXfab-Decorated Lipoplexes to Improve Delivery of siRNA Targeting C1858T PTPN22 Variant in B Lymphocytes

Autoimmune endocrine disorders, such as type 1 diabetes (T1D) and thyroiditis, at present are treated with only hormone replacement therapy. This emphasizes the need to identify personalized effective immunotherapeutic strategies targeting T and B lymphocytes. Among the genetic variants associated with several autoimmune disorders, the C1858T polymorphism of the protein tyrosine phosphatase non-receptor type 22 (PTPN22) gene, encoding for Lyp variant R620W, affects the innate and adaptive immunity. We previously exploited a novel personalized immunotherapeutic approach based on siRNA delivered by liposomes (lipoplexes) that selectively inhibit variant allele expression. In this manuscript, we improved lipoplexes carrying siRNA for variant C1858T by functionalizing them with Fab of Rituximab antibody (Ritux(Fab)-Lipoplex) to specifically target B lymphocytes in autoimmune conditions, such as T1D. Ritux(Fab)-Lipoplexes specifically bind to B lymphocytes of the human Raji cell line and of human PBMC of healthy donors. Ritux(Fab)-Lipoplexes have impact on the function of B lymphocytes of T1D patients upon CpG stimulation showing a higher inhibitory effect on total cell proliferation and IgM+ plasma cell differentiation than the not functionalized ones. These results might open new pathways of applicability of Ritux(Fab)-Lipoplexes, such as personalized immunotherapy, to other autoimmune disorders, where B lymphocytes are the prevalent pathogenic immunocytes.

Automated summary

Autoimmune endocrine disorders such as type 1 diabetes and thyroiditis currently rely on hormone replacement therapy, highlighting the need for personalized immunotherapeutic strategies targeting T and B lymphocytes. This study improved upon a personalized immunotherapeutic approach by functionalizing liposomes with Rituximab antibody, specifically targeting B lymphocytes in conditions like type 1 diabetes. The results showed that these functionalized liposomes had a significant inhibitory effect on the function of B lymphocytes, suggesting potential applicability for personalized immunotherapy in other autoimmune disorders where B lymphocytes are the primary pathogenic immune cells.