A mannose-binding lectin from Helianthus annuus termed Helja recognizes gp120 from HIV-1 promoting a pro-inflammatory phenotype in dendritic cells

CHOP, MAIA; RADICIONI MELISA; DEL RÍO MARIANELA; SABATTÉ JUAN; REGENTE MARIANA; RODRIGUEZ RODRIGUES, CHRISTIAN

Mar del Plata

Congreso; Reunión Anual de Sociedades de Biociencias (SAIC-SAB-AAFE-AACyTAL); 2023

SAIC-SAB-AAFE-AACyTAL

Background: Helja is a mannose-binding lectin isolated from sunflower seeds. HIV-1 envelope glycoprotein (gp120) contains high-mannose and complex N-glycans. The aim of this work is to analyze if Helja recognizes gp120 leading to a pro-inflammatory response. Methods: Helja was purified from Helianthus annuus seeds by D-mannose-agarose affinity chromatography. Molecular interaction between Helja and recombinant gp120 was evaluated by dot blot and ligand blot approaches using anti-Helja antibodies. Bone marrow dendritic cells (BMDCs) were cultured in complete RPMI medium supplemented with FLT3-L. Cellular activation state was evaluated in Helja-treated BMDCs by measuring membrane proteins (CD86, CD40, MHCII, CD11c, CD80, Clec9a, Ly6G, SIPRα, CD103, SiglecH) by FACS. BMDCs cytokine profile was analyzed by RT-qPCR (il-6, tnf-α, il-12, inf-α tgf-b, and il-10). Results: Helja interaction with gp120 was measured in dot blot assays by immunodetection and confirmed in ligand blot assays detecting a signal of 95 kDa. Also, Helja (10 µg/ml) was added to BMDC cultures inducing an upregulation of MHCII, MHCI, CD80, CD40 and CD86 (n=3,**p