Heterologous abs from llama and chicken egg yolk to prevent rotavirus diarrhea

描述（由申请人提供）：我们将探索口服不同的被动异源抗体（Abs）、鸡卵黄IgY和羊驼重链抗体衍生的重组VHH片段，作为轮状病毒（RV）腹泻预防和治疗的替代或补充方法。被动治疗将在猪和小牛两种新生RV疾病动物模型中进行评估。该研究将同时在美国俄亥俄州立大学Saif博士的实验室（Gn猪）和阿根廷INTA与Parreqo博士合作（新生儿小牛）进行，作为NIAID的延伸，NIH拨款号。R01 ai033561-11（6/25/03-12/31/08）。Saif博士和dr。Parreqo和Fernandez （INTA）促进了阿根廷主要农产品牛的RV感染诊断（分析、试剂）和治疗（疫苗）最先进技术的转让。通过这项提议，我们将继续合作，通过探索新的免疫学概念（被动异源抗体对RV活性抗体反应的影响）和新的治疗方法（被动IgY，VHH）来改善反刍动物和人类的健康，这两种新生动物易患RV腹泻：具有正常肠道微生物菌群的常规小牛和缺乏微生物菌群的Gn猪。外国合作者将生产抗体（羊驼vhh，鸡蛋IgY，牛IgG）并进行小牛实验。INTA的学生将在Saif博士的实验室接受培训，学习新的免疫程序（流式细胞术等），并与专门的动物疾病模型（Gn猪）一起工作，同时提高与粘膜免疫相关的分析技能。因此，INTA团队将接受与Gn猪粘膜免疫反应研究相关的试剂和培训，这些试剂和培训将应用于犊牛实验。我们的合作也将进一步提高INTA在拟议领域作为基础和转化研究卓越中心的声誉。从羊驼中提取重组抗体，将使这种本土动物的可持续应用，用于开发用于治疗这种疾病和其他疾病的VHH抗体。这将有利于骆驼畜牧业生产，也有利于阿根廷或其他从事骆驼生产的南美洲较贫穷发展中国家（秘鲁、玻利维亚等）的区域微观经济。这些异源被动抗体治疗将对早产儿或RV严重爆发环境（医院、日托中心、发展中国家农村地区）的补充药物产生深远影响。它们是在不可能、太昂贵、太迟使用RV疫苗的情况下减少RV腹泻严重程度和死亡的补充治疗；或者治疗婴儿和反刍动物疫苗失败的病例。公共卫生相关性：轮状病毒（RV）是世界范围内幼龄动物和人类婴儿腹泻的主要原因。目前，动物的RV预防是基于灭活RV母疫苗来增强被动免疫，而最近已批准用于人类婴儿的RV减毒活疫苗。疫苗的效力和安全性仍不清楚，特别是在发展中国家。我们将探索口服不同的被动异种抗体、羊驼重链抗体片段（VHH）和卵黄IgY作为控制动物和人类RV腹泻的替代/补充方法，在两种轮状病毒感染和疾病的动物模型中，猪和新生小牛（Gn）。如果我们的异源被动抗体（Ab）策略有效，这些治疗方法作为早产儿或RV严重爆发环境（医院、日托中心、发展中国家的农村地区）的补充，将产生最深远的影响，在RV疫苗不可能使用、太昂贵、太迟的情况下，通过提供一种替代的快速治疗来减少RV腹泻的严重程度和死亡；或者治疗疫苗失败的病例。

DESCRIPTION (provided by applicant): We will explore oral administration of different passive heterologous antibodies (Abs), chicken egg yolk IgY and recombinant VHH fragments derived from llama heavy chain Abs as an alternative or complementary approach to vaccination for rotavirus (RV) diarrhea prevention and treatment. The passive treatments will be evaluated in two neonatal animal models of RV disease, gnotobiotic (Gn) pigs and calves. The research will be done simultaneously, in the US at Dr. Saif's Lab, The Ohio State University (Gn pig) and in Argentina, at INTA in collaboration with Dr. Parreqo (neonatal calves), as an extension of NIAID, NIH Grant No. R01 AI033561-11 (6/25/03-12/31/08). The ongoing and long-term collaboration between Dr. Saif and Drs. Parreqo and Fernandez (INTA) has facilitated transfer of state-of-the-art technology for diagnosis (assays, reagents) and treatments (vaccines) for RV infections of cattle, the major agricultural commodity in Argentina. Through this proposal we continue this collaboration related to improving both ruminant and human health by exploring new immunologic concepts (impact of passive heterologous Ab on active Ab responses to RV) and new treatments (passive IgY,VHH) in two neonatal animals susceptible to RV diarrhea: conventional calves with a normal gut microbial flora and Gn pigs lacking the microbial flora. The foreign collaborator will produce the Abs (llama VHH, chicken egg IgY, bovine IgG) and conduct the calf experiments. Students from INTA will be trained in Dr. Saif's lab to learn newer immunologic procedures (flow cytometry, etc) and work with specialized animal disease models (Gn pigs) as well as to enhance analytical skills related to mucosal immunity. Thus the INTA team will receive reagents and training related to the study of mucosal immune responses in Gn pigs that will be applied to the calf experiments. Our collaboration will also further INTA's reputation as a Center of Excellence for basic and translational studies in the proposed areas. Development of recombinant Abs from llamas, will enable a sustainable application of this native animal for the development of VHH Ab applied to treat this and other diseases. This will benefit llama livestock production, as well as regional micro-economies for Argentina or other poorer developing countries of South America (Peru, Bolivia, etc) engaged in Camelid production. These heterologous passive Ab treatments would have profound impacts as supplements for premature infants or in severe RV outbreak settings (hospitals, day care centers, rural areas of developing countries). They represent complementary treatments to reduce RV diarrhea severity and deaths under conditions where RV vaccine use is not possible, too expensive, too late; or to treat cases of vaccine failure for both infants and ruminants. PUBLIC HEALTH RELEVANCE: Rotavirus (RV) is the leading cause of diarrhea in young animals and human infants, worldwide. Currently, RV prevention in animals is based on inactivated RV maternal vaccines to enhance passive immunity, whereas live-attenuated RV vaccines have been recently approved for human infants. Vaccine efficacy and safety remain unclear especially in developing countries. We will explore oral administration of different passive heterologous antibodies, llama heavy chain antibody fragments (VHH) and chicken egg yolk IgY as alternative/complementary approaches to control RV diarrhea for both animals and humans, in two animal models of rotavirus infection and disease gnotobiotic (Gn) pigs and neonatal calves. If our heterologous passive antibody (Ab) strategies are effective, these treatments would have the most profound impacts as supplements for premature infants or in severe RV outbreak settings (hospitals, day care centers, rural areas of developing countries), by providing an alternative rapid treatment to reduce RV diarrhea severity and deaths under conditions where RV vaccine use is not possible, too costly, too late; or to treat cases of vaccine failure.