Fusco Luciano S, Lopez Gisela L, Maslovski Franco, Brignone Sofía, Chaves María G, Calvete Juan J, Franco Yanet G, Hernandez David, Van de Velde Andrea, Marin Constanza, Palma Santiago, Maletto Belkys, Moron Gabriel, Leiva Laura C
Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Instituto de Química Básica y Aplicada del Nordeste Argentino (IQUIBA-NEA), CP3400 Corrientes, Argentina.
Facultad de Ciencias Exactas Naturales y Agrimensura, Universidad Nacional del Nordeste, CP3400 Corrientes, Argentina.
Trans R Soc Trop Med Hyg. 2025 Sep 2;119(9):1070-1083. doi: 10.1093/trstmh/trae129.
The WHO states that antivenom is the only safe and effective treatment to neutralize snake venom. Snakebite antivenom typically involves horse hyperimmunization with crude venom and Freund's adjuvant.
In the current work, we analyzed the ascorbyl palmitate liquid crystal structure with snake protein or PLA2, the carrier charge capacity, and we evaluated the immune response induced by the enzyme P9a(Cdt-PLA2) formulated in a nanostructure using CpG-ODN, determining the titer of IgG antibodies. BALB/c mice were subcutaneously immunized on days 0, 15 and 30 with P9a(Cdt-PLA2)/CpG-ODN/Coa-ASC16 or P9a(Cdt-PLA2)/Freund's adjuvant (complete first and incomplete-booster). On day 48 the mice were sacrificed. The neutralization ability of antibodies from animals immunized with P9a(Cdt-PLA2)/CpG-ODN/Coa-ASC16 or P9a(Cdt-PLA2)/Freund's adjuvant was tested against PLA2 activity and venom lethality.
In both groups of immunized mice, the antibody titers in blood samples at the assayed time were high (approximately 1×105). The antibodies were able to neutralize P9a(Cdt-PLA2) activity in vitro and lethality in vivo. Microscopic analysis showed that P9a(Cdt-PLA2)/CpG-ODN/Coa-ASC16 produces minimal damage at injection sites compared with Freund's adjuvant.
The Coa-ASC16/CpG-ODN formulation shows promise as a safe and effective adjuvant against crotalic PLA2, inducing a strong humoral response and reducing local tissue damage compared with Freund's adjuvant.
世界卫生组织指出，抗蛇毒血清是中和蛇毒的唯一安全有效的治疗方法。蛇咬伤抗蛇毒血清通常涉及用粗毒液和弗氏佐剂对马进行超免疫。
在当前的研究中，我们分析了抗坏血酸棕榈酸酯与蛇蛋白或磷脂酶A2（PLA2）的液晶结构、载体电荷容量，并评估了使用CpG-寡脱氧核苷酸（ODN）在纳米结构中配制的酶P9a（Cdt-PLA2）诱导的免疫反应，确定了IgG抗体的滴度。在第0、15和30天，用P9a（Cdt-PLA2）/CpG-ODN/Coa-ASC16或P9a（Cdt-PLA2）/弗氏佐剂（首次为完全佐剂，加强免疫为不完全佐剂）对BALB/c小鼠进行皮下免疫。在第48天处死小鼠。测试用P9a（Cdt-PLA2）/CpG-ODN/Coa-ASC16或P9a（Cdt-PLA2）/弗氏佐剂免疫的动物产生的抗体对PLA2活性和毒液致死性的中和能力。
在两组免疫小鼠中，在测定时间采集的血样中的抗体滴度都很高（约1×10⁵）。这些抗体能够在体外中和P9a（Cdt-PLA2）的活性，并在体内中和致死性。显微镜分析表明，与弗氏佐剂相比，P9a（Cdt-PLA2）/CpG-ODN/Coa-ASC16在注射部位产生的损伤最小。
Coa-ASC16/CpG-ODN制剂有望作为一种安全有效的抗响尾蛇PLA2佐剂，与弗氏佐剂相比，能诱导强烈的体液反应并减少局部组织损伤。
