Characterization of leukocyte functions from cattle with leukocyte adhesion deficiency (BLAD) and prevalence of BLAD-carrier in Holstein cattle in Japan

白细胞粘附缺陷 (BLAD) 牛的白细胞功能特征以及日本荷斯坦牛中 BLAD 携带者的患病率

• 批准号: 07660426
• 负责人: NAGAHATA Hajime
• 金额: $ 0.96万
• 依托单位: RAKUNO GAKUEN UNIVERSITY,FACULTY OF VETERINARY MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660426/
• 关键词: BLAD; CD18; bovine immunodeficiency; leukocyte functions; BLAD-carrier; disease control; 好中球機能; DNA-PCR; キャリャ-率; 遺伝子頻度; CD18; 免疫組織検索; Fcリセプター; BLAD-キャリャ-

The above-entitled study was performed and the results were summarized as follows :Severe bone resorption, osteolysis and severe progressive periodontitis in submandibula due to dysfunction of leukocytes in heifers with BLAD were demonstrated by radiographic examination. Immunohistochemical analysis of lymph nodes using anti-CD18 antibody demonstrated that CD18-positive cells were not found on those from the BLAD-heifer.Fc receptor expression, cytoplasmic Ca^<2+> signaling, chemiluminescent (CL) response, and electron spin resonance (ESR) combined with spin trapping of blood mononuclear phagocytes from control heifers and a heifer with BLAD were evaluated to elucidate the relationships between complement receptor type 3 (CR3) and Fc receptor expression and their functional responses. Fc receptors on mononuclear phagocytes from the heifer with BLAD were enhanced, and their cytoplasmic Ca^<2+> singnaling, CL responses, and ESR-spin trapping when stimulated with aggregated IgG (Agg-IgG) o … More psonized zymosan (OPZ) appeared to be associated with enhanced Fc receptors.The adhesiveness of control and CD18-deficient bovine neutrophils on culture plates precoated with collagen I,collagen IV,fibronectin and laminin was measured to evaluate the possible factors for adherence to extracellular matrices. The adhesiveness activities of CD18-deficient neutrophils to collagen I,collagen IV and fibronectin were significantly diminished ; however, similar adhesion to laminin was observed in CD18-deficient neutrophils and control neutrophils.Biosynthesis of CD11/CD18 in bovine leukocytes, intracellular Ca^<2+> signaling, CL responses and membrane fluidity of neutrophils from control heifers and a BLAD-heifer were measured. The synthesis of CD11/CD18 complex was clearly detected in leukocytes from a normal heifer, but not in a BLAD-heifer. The ttransient phase of increased Ca^<2+> was clearly detected in neutrophils from a BLAD-heifer stimulated with OPZ,Agg-IgG,whereas the sustained phase was deficient or significantly decreased compared with controls. No differences in membrane fluidity were detected between neutrophils derived from control and BLAD-cattle.Blood samples from 796 Holstein dairy cows in 20 herds from 6 districts in Japan were examined to determine whether they were BLAD-free, BLAD carriers, or BLAD-affected by use of DNA-PCR analysis, BLAD-carrier rates ranged from 0 to 23.5% and the mean BLAD-carrier rate was 8.1% (70/796). Approximately 35% of the dairy herds had a prevalence of more than 10% of BLAD-carriers. The rate of the occurrence of BLAD in Japanese Holstein dairy cattle was estimated to be 0.16% at birth. A program for the eradication of the gene encoding impaired CD18 for BLAD in Holstein cattle is slowly being enacted in Japan.These results demonstrate that pathophysiological changes of BLAD are closely associated with the deficiency of CD18 expression on BLAD-leukocytes. This study provide a valuable information for the control of BLAD in Japanese Holstein cattle. Less

进行了上述研究，结果总结如下：BLAD小母牛的白细胞功能障碍导致下颌下严重骨吸收、骨质溶解和严重进行性牙周炎，通过使用抗CD18的淋巴结免疫组织化学分析来证明。抗体证明在来自BLAD小母牛的细胞中未发现CD18阳性细胞。Fc受体表达、细胞质Ca ^ 2+ 信号传导、对来自对照小母牛和患有 BLAD 的小母牛的血液单核吞噬细胞的化学发光 (CL) 反应和电子自旋共振 (ESR) 结合自旋捕获进行评估，以阐明补体受体 3 型 (CR3) 和 Fc 受体表达及其功能之间的关系患有BLAD的小母牛的单核噬菌体上的Fc受体反应增强，并且它们的细胞质Ca ^ 2+ 信号传导、CL细胞反应和当用聚集的 IgG (Agg-IgG) o … 更多 psonized zymosan (OPZ) 刺激时，ESR-自旋捕获似乎与增强的 Fc 受体相关。对照和 CD18 缺陷的牛中性粒细胞在预涂胶原蛋白 I、胶原蛋白的培养板上的粘附性IV、测量纤连蛋白和层粘连蛋白以评价CD18缺陷细胞粘附细胞外基质的可能因素。中性粒细胞对 I 型胶原、IV 型胶原和纤连蛋白的粘附显着减少；然而，在 CD18 缺陷型中性粒细胞和对照中性粒细胞中观察到与层粘连蛋白相似的粘附。牛白细胞中 CD11/CD18 的生物合成、细胞内 Ca^2+ 信号传导、CL 反应并测量了对照小母牛和 BLAD 小母牛的中性​​粒细胞的膜流动性。在来自正常小母牛的白细胞中清楚地检测到CD11/CD18复合物，但在BLAD小母牛中没有。在来自用OPZ、Agg-IgG刺激的BLAD小母牛的嗜中性粒细胞中清楚地检测到增加的Ca 2+ 的瞬时阶段。 ，而与对照相比，持续相不足或显着降低。对照和 BLAD 牛的中性粒细胞之间没有检测到膜流动性差异。研究人员对日本 6 个地区 20 群 796 头荷斯坦奶牛的样本进行了 DNA-PCR 分析，以确定它们是否无 BLAD、携带 BLAD 或受 BLAD 影响，BLAD 携带率范围为 0 至 23.5%奶牛群的平均 BLAD 携带率为 8.1% (70/796)。 BLAD 携带者的患病率超过 10% 日本荷斯坦奶牛出生时 BLAD 的发生率为 0.16% 一项旨在消除荷斯坦牛中编码受损 CD18 的基因的计划。这些结果表明BLAD的病理生理变化与BLAD白细胞上CD18表达缺陷密切相关。本研究提供了有价值的信息。用于控制日本荷斯坦牛的 BLAD。

项目成果

Nagahata,H.,et al.: "Biosynthesis of β_2 integrin, calcium signalling and functional respences of borine nectrophils." Research in Veterinary Science. 61. 95-101 (1996)

Nagahata, H., et al.：“β_2 整合素的生物合成、钙信号传导和硼嗜性粒细胞的功能反应。兽医科学研究”61. 95-101 (1996)。

桑原・郷司・稲波・樋口・永幡: "白血球粘着蛋白質欠損症ホルスタイン牛(BLAD)好中球を用いた活性酸素生成・情報伝達機構の研究" 編集 内海英雄,吉川敏一(日本医学館)磁気共鳴と医学(8刊), 5(109-113) (1997)

Kuwabara、Goji、Inami、Higuchi、Nagahata：“利用白细胞粘附蛋白缺陷 (BLAD) 荷斯坦奶牛的中性粒细胞研究活性氧的产生和信息传递机制”编辑：Hideo Utsumi、Toshikazu Yoshikawa（日本医学会）磁共振与医学（第 8 版），5(109-113) (1997)

H.Nagahata: "Neutrophil and mononuclear phagocyte functions from cattle with leucocyte adhesion deficiency (BLAD) and prevalence of BLAD-carrier in Holstein cattle in Japan." Proceedings of XIX World Buiatrics Congress. 2. 489-492 (1996)

H.Nagahata：“白细胞粘附缺陷 (BLAD) 牛的中性粒细胞和单核吞噬细胞功能以及日本荷斯坦牛中 BLAD 携带者的流行。”

Nagahata,H.,et al.: "Functional characteristics of enhanced Fc reuptor expression of β_2-integrin deficient bovire monocytes." Microbiology and Immunology. 40. 389-395 (1996)

Nagahata, H., et al.：“β_2-整合素缺陷的牛单核细胞的 Fc 受体表达增强的功能特征。微生物学和免疫学”40. 389-395 (1996)。

H.Nagahata: "Biosynthesis of B_2 integrin, intracellular calcium signalling and functional responses of normal and CD18-deficient bovine neutrophils." Res.Vet.Sci.61. 95-101 (1996)

H.Nagahata：“B_2 整合素的生物合成、细胞内钙信号传导以及正常和 CD18 缺陷牛中性粒细胞的功能反应。”