重组人碱性成纤维细胞生长因子在猕猴和大鼠中中和抗体的产生及其在肾脏沉积

Production of neutralizing antibodies to recombinant human basic fibro blast growth factor and its renal deposition in rhesus monkeys and rats

目的：在猕猴和大鼠中进行重组人碱性成纤维细胞生长因子（ｒｈｂ Ｆ Ｇ Ｆ）的重复给药毒性试验时，观察给药后抗 ｒｈｂ Ｆ Ｇ Ｆ 抗体的产生和抗体的中和性， 以及抗体在肾脏的沉积。方法：猕猴分生理盐水对照组、ｒｈｂ Ｆ Ｇ Ｆ ４ μｇ·ｋｇ－ １·ｄ－ １、２０ μｇ·ｋｇ－ １·ｄ－ １和 １００ μｇ·ｋｇ－ １·ｄ－ １ 共４ 组。大鼠分生理盐水对照组、ｒｈｂ Ｆ Ｇ Ｆ ８ μｇ·ｋｇ－ １·ｄ－ １、３２ μｇ·ｋｇ－ １·ｄ－ １和 １２８ μｇ·ｋｇ－ １·ｄ－ １ 共４ 组。采用肌肉注射，连续给药 ３０ ｄ。利用 Ｅ Ｌ Ｉ Ｓ Ａ 法检测血清中抗 ｒｈｂ Ｆ Ｇ Ｆ 的抗体。出现抗体时再用 Ｎ Ｉ Ｈ３ Ｔ３ 细胞株检测抗体的中和性并进行肾组织的免疫组织化学检查。结果：在给药期内，各组猕猴血清内未能检测到抗 ｒｈｂ Ｆ Ｇ Ｆ 抗体。给药 １５ ｄ 后，在大鼠的 ｒｈｂ Ｆ Ｇ Ｆ ３ 个剂量组中可检测出抗ｒｈｂ Ｆ Ｇ Ｆ抗体，并且这种抗体具有抑制ｒｈｂ Ｆ Ｇ Ｆ促进 Ｎ Ｉ Ｈ３ Ｔ３ 细胞增殖的作用。肾脏组织切片的免疫组化结果显示，用药组大鼠肾小球基底膜有抗体沉积。结论：在猕猴上适合进行 ｒｈｂ Ｆ Ｇ Ｆ重复给药毒性试验。中和抗体的产生及其在肾小球基底膜中沉积可?

Objective: To observe production of antibody to rhbFGF, its neutralizing activity and deposition in kidney in conducting toxicity study on repeated doses of recombinant human basic fibroblast growth factor (rhbFGF) in normal rhesus monkeys and Wistar rats. Methods: The monkeys were treated with i.m. rhbFGF at dosages of 4, 20 and 100 μg·kg -1 ·d -1 for 30 days, and the dosages in rats were 8, 32 and 128 μg·kg -1 ·d -1 , respectively. Anti rhbFGF antibodies in sera were examined using ELISA and the neutralizing activities of antibodies were evaluated for their ability to inhibit the proliferation of rhbFGF stimulated NIH3T3 cells. Results: No antibody to rhbFGF was found in rhesus monkeys during experimental period. After 15 days of treatment, anti rhbFGF antibodies were detected in sera of rhbFGF treated rats. Higher titers of anti rhbFGF antibodies developed after 20 and 25 days of treatment. The antibodies inhibited the bFGF stimulated proliferation of NIH3T3 cells. Immunohistochemical studies exhibited glomerular deposition of anti rhbFGF antibodies. Conclusion: The results indicate that the rhesus monkey is suitable for safety assessment of rhbFGF and the production of neutralizing antibodies in rats may be involved in the renal toxicity of rhbFGF.