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Changes of iron and iron-binding proteins in calves infected with small-sized piroplasma

感染小型焦原体的犊牛体内铁及铁结合蛋白的变化

基本信息

批准号：
06660411
负责人：
WATANABE Kiyotaka
金额：
$ 1.28万
依托单位：
KITASATO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06660411/
关键词：
small-sized piroplasma Theileria sergenti serum iron iron-metabolism transferrin transferrin saturation lactoferrin ferritin

项目摘要

1. Ferritin (Ft), transferrin (Tf), and lactoferrin (Lf) were highly purified frombovine spleen, serum, and milk, respectively, and rabbit antisera to each of them were prepared. Although Tf and Lf were immunologically quite different from each other in their native forms, they became cross-reactive when denatured.2. Sandwich ELISA was developed for measuring each iron-binding protein using affinity-purified antibodies.(1) Mean serm Ft concentration of healthy non-pregnant cows was 46<plus-minus>28ng/ml (n=69). (2) As heparin stimulated neutrophils, resulting in Lf release, and as Lf was released from the cells during coagulation, it was necessary to use EDTA plasma for measuring blood Lf. Mean plasma Lf and serum Tf concentrations of healthy non-pregnant cows were 169<plus-minus>63ng/ml (n=16) and 2.86([SY.+-.[)0.43mg/ml (n=72), respectiv3. Holstein female calves aged 5 to 6 months were naturally infected with Theileria sergenti by grazing them in the pasture polluted with the protozoa, and were examined for the changes of serum iron and iron-binding proteins Tf, Lf, and Ft after infection. (1) As the anemia of infected calves progressed, the levels of serum iron and Tf saturation increased over 250mug/dl and 90%, respectively, while serum Tf concentration gradually decreased. Tfa/Tfb ratios of infected calves were 2-fold higher than those of control animals, showing the qualitative change of Tf after infection. (2) With the progress of anemia, serum Ft concentration gradually increased, and erythrocyte Ft content drastically increased in accordance with the increment of MCV and was more than 200 times that of the control. Mononuclear cell Ft content of the infected cow was more than 6 times that of the uninfected cattle. (3) Plasma Lf level of the infected calf decreased to one third of that of the control.

1.分别从牛脾、血清和乳汁中分离纯化铁蛋白(Ft)、转铁蛋白(Tf)和乳铁蛋白(Lf)，制备兔抗血清。尽管天然形式的TF和LF在免疫学上彼此有很大的不同，但变性后它们会产生交叉反应。用亲和纯化抗体建立双抗体夹心ELISA法测定各铁结合蛋白：(1)健康非妊娠奶牛血清Ft平均浓度为46-28 ng/ml(n=69)。(2)由于肝素刺激中性粒细胞，导致LF的释放，以及在凝血过程中细胞释放LF，因此用EDTA血浆测定血中的LF是必要的。健康非妊娠奶牛的平均血浆LF值和血清TF值分别为169±63 ng/ml(n=16)和2.86±0.43 mg/ml(n=72)。用5~6月龄荷斯坦母犊在被原虫污染的草场放牧自然感染瑟氏泰勒氏菌，检测感染后血清铁及铁结合蛋白Tf、Lf、Ft的变化。(1)随着感染犊牛贫血的加重，血清铁水平和血清转铁蛋白饱和度分别升高到250µg/dl和90%以上，而血清转铁蛋白浓度逐渐降低。感染后TFA/TFB比值较对照高2倍，说明感染后Tf发生了质的变化。(2)随着贫血程度的加重，血清Ft浓度逐渐升高，红细胞Ft含量随MCV升高而急剧升高，为对照组的200多倍。感染牛的单个核细胞Ft含量是未感染牛的6倍以上。(3)感染组犊牛血浆LF值降至对照组的1/3。