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Research Article: Dual targeting of CD244 and CD2 by a viral-immunoevasin-guided engineered CD48-Fc mutant for T- and NK-cell modulation

Date Published: 2026-09-01

Abstract:
Viruses encode immunoevasins that subvert host co-signaling pathways. CD244:CD48 and CD2:CD58 interactions are critical for NK- and T-cell effector responses. We previously identified a herpesvirus-encoded soluble CD48 homolog that functions as a decoy ligand for CD244 and dampens NK-cell activation. Here, we show that this viral protein, A43, also recognizes CD2, and we used its structural features to engineer human CD48-Fc variants with dual CD244/CD2 specificity. The optimized mutein, C5-Fc, which incorporates twelve A43-guided substitutions predicted by structure-based modeling to enhance interface packing and electrostatic complementarity, resulted in high-avidity binding and competitively displacement of native ligands. Fc-silenced C5-Fc (C5-Fc # ) disrupted NK- and T-cell conjugate formation and reduced CD244- and CD2-dependent cytotoxicity. It also interfered with the maturation of the antigen-dependent immunological synapse and inhibited T-cell proliferation and the secretion of pro-inflammatory cytokines. These data establish C5-Fc # as a rationally engineered dual-specificity inhibitor with translational potential for autoimmune diseases.

Introduction:
Viruses have evolved sophisticated mechanisms to subvert antiviral immunity through the expression of immunomodulatory proteins ( 1 , 2 ). Knowledge gained from these potent immunoevasins is key to understand viral pathogenesis and can guide the development of antiviral and immunomodulatory therapeutics. Many viral strategies rely on host genes captured through horizontal gene transfer ( 3 ). Over evolutionary time, such genes have diversified to produce molecules that mimic or inhibit innate and adaptive immune…

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