Bobeck Elizabeth A, Hellestad Erica M, Sand Jordan M, Piccione Michelle L, Bishop Jeff W, Helvig Christian, Petkovich Martin, Cook Mark E
Animal Sciences Department, University of Wisconsin-Madison, Madison, WI 53706.
OPKO Health, Renal Division, Markham, Ontario, Canada L3R 6H3.
Poult Sci. 2015 Jun;94(6):1128-37. doi: 10.3382/ps/pev085. Epub 2015 Mar 29.
Hyperimmunized hens are an effective means of generating large quantities of antigen specific egg antibodies that have use as oral supplements. In this study, we attempted to create a peptide specific antibody that produced outcomes similar to those of the human pharmaceutical, sevelamer HCl, used in the treatment of hyperphosphatemia (a sequela of chronic renal disease). Egg antibodies were generated against 8 different human intestinal sodium-dependent phosphate cotransporter 2b (NaPi2b) peptides, and hNaPi2b peptide egg antibodies were screened for their ability to inhibit phosphate transport in human intestinal Caco-2 cell line. Antibody produced against human peptide sequence TSPSLCWT (anti-h16) was specific for its peptide sequence, and significantly reduced phosphate transport in human Caco-2 cells to 25.3±11.5% of control nonspecific antibody, when compared to nicotinamide, a known inhibitor of phosphate transport (P≤0.05). Antibody was then produced against the mouse-specific peptide h16 counterpart (mouse sequence TSPSYCWT, anti-m16) for further analysis in a murine model. When anti-m16 was fed to mice (1% of diet as dried egg yolk powder), egg yolk immunoglobulin (IgY) was detected using immunohistochemical staining in mouse ileum, and egg anti-m16 IgY colocalized with a commercial goat anti-NaPi2b antibody. The effectiveness of anti-m16 egg antibody in reducing serum phosphate, when compared to sevelamer HCl, was determined in a mouse feeding study. Serum phosphate was reduced 18% (P<0.02) in mice fed anti-m16 (1% as dried egg yolk powder) and 30% (P<0.0001) in mice fed sevelamer HCl (1% of diet) when compared to mice fed nonspecific egg immunoglobulin. The methods described and the findings reported show that oral egg antibodies are useful and easy to prepare reagents for the study and possible treatment of select diseases.
超免疫母鸡是产生大量具有口服补充剂用途的抗原特异性卵抗体的有效手段。在本研究中,我们试图制备一种肽特异性抗体,其产生的结果与用于治疗高磷血症(慢性肾病的后遗症)的人类药物盐酸司维拉姆相似。针对8种不同的人肠钠依赖性磷酸转运蛋白2b(NaPi2b)肽产生卵抗体,并筛选hNaPi2b肽卵抗体抑制人肠Caco-2细胞系中磷酸盐转运的能力。针对人肽序列TSPSLCWT产生的抗体(抗-h16)对其肽序列具有特异性,与已知的磷酸盐转运抑制剂烟酰胺相比,可将人Caco-2细胞中的磷酸盐转运显著降低至对照非特异性抗体的25.3±11.5%(P≤0.05)。然后针对小鼠特异性肽h16对应物(小鼠序列TSPSYCWT,抗-m16)产生抗体,以便在小鼠模型中进行进一步分析。当将抗-m16喂给小鼠(以干蛋黄粉形式占饮食的1%)时,使用免疫组织化学染色在小鼠回肠中检测到卵黄免疫球蛋白(IgY),并且卵抗-m16 IgY与商业山羊抗-NaPi2b抗体共定位。在一项小鼠喂养研究中,确定了与盐酸司维拉姆相比,抗-m16卵抗体降低血清磷酸盐的有效性。与喂食非特异性卵免疫球蛋白的小鼠相比,喂食抗-m16(以干蛋黄粉形式占1%)的小鼠血清磷酸盐降低了18%(P<0.02),喂食盐酸司维拉姆(占饮食的1%)的小鼠血清磷酸盐降低了30%(P<0.0001)。所描述的方法和所报告的发现表明,口服卵抗体是用于特定疾病研究和可能治疗的有用且易于制备的试剂。