Differential Modulation of SARS-CoV-2 Infection by Complement Factor H and Properdin

Uday Kishore^1*Praveen Mathews Varghese²Chandan Kumar³Susan Idicula-Thomas³Martin Mayora Neto⁴Anthony George Tsolaki⁵Pretty Ponnachan⁶Khaled Masmoudi⁶Basel K Al-Ramadi⁶Manu Vatish⁷Taruna Madan³Nigel James Temperton⁴Nazar Beirag²

• ¹Biosciences, United Arab Emirates University, Al-Ain, United Arab Emirates
• ²Umea University, Umea, Sweden
• ³ICMR NIRRCH, Mumbai, India
• ⁴University of Kent and Greenwich, Kent, United Kingdom
• ⁵Brunel University of London, London, United Kingdom
• ⁶UAE University, Al Ain, United Arab Emirates
• ⁷University of Oxford, Oxford, United Kingdom

An unbalanced immune response and excessive inflammation are the major hallmarks of severe SARS-CoV-2 infection. The dysregulation of the complement system is known as a crucial factor in the immunopathology of SARS-CoV-2 infection. Complement alternative pathway has been linked to the excessive inflammation in severe SARS-CoV-2 infection in which decreased and elevated levels of factor H (FH) and properdin (FP), respectively were observed. The current study investigated the potential immunomodulatory roles of FP and FH against SARS-CoV-2 infection. The interactions between FH and FP and the SARS-CoV-2 spike (S) and its receptor binding domain (RBD) were evaluated using direct ELISA. The cell binding and luciferase based viral entry assays utilising S protein expressing lentiviral pseudotypes were used to assess the effects of FH, FP, and recombinant thrombospondin repeats 4 and 5 (TSR4+5) on SARS-CoV-2 cell entry. Using RT-qPCR, we also assessed the effects of FH and FP in the cytokine response induced by SARS-CoV-2 pseudotypes. FH and FP bound to RBD and SARS-CoV-2 S protein. The treatment with FP or TSR4+5 enhanced cell binding and entry of SARS-CoV-2 pseudotypes that was administered in A549 cells expressing human ACE2 and TMPRSS2 (A549-hACE2+TMPRSS2 cells). FP increases the affinity between host ACE2 and SARS-CoV-2, according to in silico analysis. In A549-hACE2+TMPRSS2 cells, the effect of FP on viral cell entry and binding was counteracted by anti-FP antibody treatment. SARS-CoV-2 lentiviral pseudotypes' cell entry and binding were decreased by FH. The A549-hACE2+TMPRSS2 cells that were challenged with SARS-CoV-2 alphaviral pseudotypes (expressing spike, envelope, nucleocapsid, and membrane proteins) pre-treated with FP or TSR4+5 showed an upregulation of pro-inflammatory cytokine transcripts, including NF-B and IL-1β, IL-8, IL-6, TNF-α, IFN-α, and RANTES. Contrary to this, the expression of these pro-inflammatory cytokines was downregulated by FH treatment. FH treatment decreased S protein-mediated NF-κB activation, but FP treatment enhanced it in A549-hACE2+TMPRSS2 cells. These results imply that FH may function as a SARS-CoV-2 cell entry and binding inhibitor, reducing the inflammatory response linked to infection independently of complement activation. FP could aid cell viral entry and binding and aggravate hyperinflammation that might contribute to the severity of the infection.