Bifunctional antibody fragment-based fusion proteins for the targeted elimination of pathogenic T-cell subsets.
Pathogenic effector T cells are key contributors to autoimmune diseases such as systemic lupus erythematosus (SLE). General inhibition of T cells using, e.g., methotrexate, prednisolone, or TNF blockers, has prominent therapeutic effects frequently at the cost of severe long-term side effects and toxicity. Therefore, targeted strategies that can selectively inhibit or eliminate pathogenic T cells are sought after as a new approach to safely block perpetual inflammatory T-cell responses and inhibit concomitant progressive tissue destruction. Of particular interest in this respect is the use of the so-called single-chain fragments of variable region (scFv) antibody fragments for the targeted reactivation of Fas-dependent activation-induced cell death (AICD). Recently, we demonstrated that a recombinant fusion protein comprising a T-cell-targeted anti-CD7 scFv antibody fragment genetically fused to soluble FasL (sFasL) can eliminate synovial fluid T cells in the absence of activity toward resting peripheral blood cells. Here, we describe a detailed protocol for construction and preclinical evaluation of such scFv:FasL fusion proteins that may be used to selectively eliminate pathogenic immune cells.
Department of Surgery, Translational Surgical Oncology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
Wijnand
W
Helfrich
Edwin
E
Bremer
D000818:Animals; D018033:Antibodies, Bispecific / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D016466:CHO Cells; D003412:Cricetulus; D053222:Fas Ligand Protein / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D008180:Lupus Erythematosus, Systemic / Q000276:immunology / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding / Q000276:immunology; D011993:Recombinant Fusion Proteins; D016176:T-Lymphocyte Subsets / Q000276:immunology* / Q000378:metabolism
D018033:Antibodies, Bispecific; D053222:Fas Ligand Protein; D007128:Immunoglobulin Fragments; D011993:Recombinant Fusion Proteins