Komine Ken-ichi, Komine Yumiko, Kuroishi Toshinobu, Kobayashi Jin, Obara Yoshiaki, Kumagai Katsuo
T-Cell Research Institute, In Kyouitsu Seiyaku Corp, Tokyo, Japan.
J Vet Med Sci. 2005 Jul;67(7):667-77. doi: 10.1292/jvms.67.667.
We have identified various lactoferrin (Lf) molecules in mastitic mammary gland secretions (MGSs), and these Lf molecules were examined for their physiological function in MG. These Lf molecules were isolated by Con A affinity chromatography, and then analyzed by various electrophoresis methods and N-terminal amino acid sequencing. The low Con A affinity Lf was found to have low molecular peptides as compared with the 86 kDa of the high Con A affinity Lf, which is usually detected in healthy MGSs. The N-terminal amino acid sequence of each of the small molecular Lfs were confirmed as fragments of 86 kDa Lf. This low Con A affinity Lf stimulated spleen adherent cells to produce more O(2)(-) than 86 kDa Lf. Furthermore, the low Con A affinity Lf showed low antibacterial activity against E. coli and S. aureus, and had decreased iron-binding capacity in comparison with 86 kDa Lf. Moreover, the 86 kDa Lf could stimulate bovine T cells or macrophages to produce IFN-gamma, IL-4, IL-6, and IL-1alpha. However low Con A affinity Lf induced the production of TNFalpha, but not physiological T cell or macrophage cytokines. It was also found that when the healthy MGs of dry cows were injected with the low Con A affinity Lf, there was an increase in polymorphonuclear cells together with TNFalpha, MCP-1, and IL-8 production. These results suggested that low Con A affinity Lf in mastitic MGSs differed from 86 kDa Lf in physiological characteristics, and, that it induced an inflammatory reaction in MGs.
我们已在患乳腺炎的乳腺分泌物（MGSs）中鉴定出多种乳铁蛋白（Lf）分子，并对这些Lf分子在乳腺中的生理功能进行了研究。这些Lf分子通过伴刀豆球蛋白A亲和层析法分离，然后用各种电泳方法和N端氨基酸测序进行分析。结果发现，与通常在健康MGSs中检测到的86 kDa高伴刀豆球蛋白A亲和性Lf相比，低伴刀豆球蛋白A亲和性Lf具有低分子量肽段。每个小分子Lf的N端氨基酸序列被确认为86 kDa Lf的片段。与86 kDa Lf相比，这种低伴刀豆球蛋白A亲和性Lf刺激脾黏附细胞产生更多的超氧阴离子（O₂⁻）。此外，低伴刀豆球蛋白A亲和性Lf对大肠杆菌和金黄色葡萄球菌的抗菌活性较低，并且与86 kDa Lf相比，其铁结合能力下降。此外，86 kDa Lf可刺激牛T细胞或巨噬细胞产生干扰素-γ（IFN-γ）、白细胞介素-4（IL-4）、白细胞介素-6（IL-6）和白细胞介素-1α（IL-1α）。然而，低伴刀豆球蛋白A亲和性Lf可诱导肿瘤坏死因子-α（TNFα）的产生，但不能诱导生理性T细胞或巨噬细胞细胞因子的产生。还发现，当给干奶牛的健康乳腺注射低伴刀豆球蛋白A亲和性Lf时，多形核细胞数量增加，同时TNFα、单核细胞趋化蛋白-1（MCP-1）和白细胞介素-8（IL-）的产生也增加。这些结果表明，患乳腺炎的MGSs中的低伴刀豆球蛋白A亲和性Lf在生理特性上与86 kDa Lf不同，并且它可在乳腺中诱导炎症反应。
