Garmendia A E, Morgan D O, Baxt B
Molecular Biology Laboratory, USDA, ARS, Plum Island Disease Center, Greenport, New York 11944.
Immunology. 1989 Oct;68(2):265-71.
A neutralizing monoclonal antibody (nmAb) to foot-and-mouth disease virus (FMDV) was used as antibody-1 (AB1) to induce anti-idiotypic antibodies (a-IdAb) in rabbits. The rabbit a-IdAb (AB2) were isolated on protein A-Sepharose, followed by cycles of separation on idiotype and isotype affinity columns. The specificity of the AB2 for the paratope of AB1 was determined by direct binding to AB1 in solid-phase radioimmunoassay (SP-RIA), and by competition RIA (C-RIA) with virus for binding to the AB1. The AB2, termed a-2PD11, was utilized to immunize six groups of female Swiss mice at weekly intervals with either one of three formulations, in doses of 50 micrograms or 5 micrograms, given in single subcutaneous (s.c.) spots. Anti-viral antibody (AB3) was first detected by RIA at the fifth week in the 50 micrograms/dose groups, and maximum levels were reached at the sixth week in the 50 and 5 micrograms/dose groups. The AB3 levels were at least three times higher for mice given 50 micrograms doses. In addition, the AB3 were also shown to neutralize FMDV infectivity in tissue culture and in a suckling mouse protection assay. Overall, mice exhibited variable responses to immunization with AB2. In a subsequent trial, mice received multispot s.c. and footpad injections of 50 micrograms of a-2PD11 coupled to keyhole limpet haemocyanin (KLH) on a weekly basis. In these mice, AB3 was detected earlier than in mice immunized with single s.c. injections. These results support the use of a-IdAb as potential surrogates of critical determinants for FMD vaccines.
一种针对口蹄疫病毒（FMDV）的中和单克隆抗体（nmAb）被用作抗体1（AB1），以在兔体内诱导抗独特型抗体（a-IdAb）。兔a-IdAb（AB2）在蛋白A-琼脂糖凝胶上分离，随后在独特型和同种型亲和柱上进行多轮分离。通过在固相放射免疫分析（SP-RIA）中与AB1直接结合，以及通过与病毒竞争结合AB1的竞争放射免疫分析（C-RIA），确定AB2对AB1互补位的特异性。将名为a-2PD11的AB2以三种制剂之一，按50微克或5微克的剂量，每周一次皮下（s.c.）单点注射，免疫六组雌性瑞士小鼠。在50微克/剂量组中，第五周通过放射免疫分析首次检测到抗病毒抗体（AB3），在50微克/剂量组和5微克/剂量组中，第六周达到最高水平。给予50微克剂量的小鼠的AB3水平至少高三倍。此外，AB3在组织培养和乳鼠保护试验中也显示出能中和FMDV的感染性。总体而言，小鼠对用AB2免疫表现出不同的反应。在随后的试验中，小鼠每周接受多点皮下和足垫注射50微克与钥孔血蓝蛋白（KLH）偶联的a-2PD11。在这些小鼠中，检测到AB3的时间比单次皮下注射免疫的小鼠更早。这些结果支持将a-IdAb用作FMD疫苗关键决定因素的潜在替代物。
