Pathobiology of leukocyte functions from cattle with leukocyte adhesion deficiency(BLAD)and trials of bone marrow transplantation to BLAD-cattle.

白细胞粘附缺陷（BLAD）牛白细胞功能的病理学以及 BLAD 牛骨髓移植试验。

• 批准号: 09660344
• 负责人: NAGAHATA Hajime
• 金额: $ 2.05万
• 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: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09660344/
• 关键词: BLAD TRANSFUSION; BONE MARROW TRANSPLANTATION; CD18; GENETIC DISEASE; TRANSFUSION; CATTLE DISEASE

The above-entitled study was performed and the results were summarized as follows :Fc receptor for immunoglobulin G-mediated phagocytosis, superoxide production and intracellular calcium ([Ca^2+]^i) signaling of complement receptor type 3-(CR3) deficient neutrophils from a heifer with leukocyte adhesion deficiency were compared to those of control heifers. The mean phagocytic activity of IgG-coated yeasts and aggregated bovine IgG (Agg-IgG)-induced superoxide production of CR3-deficient neutrophils were 10% and 77.9%, respectively, of those of control neutrophils. The [Ca^2+]^i signals in CR3-deficient neutrophils stimulated with Agg-Igg or concanavalin A were deficient with mean peak [Ca^2+]^i concentrations of 78% and 42% of those of controls. These findings suggest that Fc receptor-mediated neutrophil functions are closely dependent on the presence of CR3 on the neutrophil cell surfaces.The emigration of leukocytes from calves with B2 integrin deficiency (BLAD) into bronchoalveolar … More spaces and scraped tissues was compared to that of normal calves. Neutrophils were found in bronchoalveolar lavage fluid from BLAD-affected calves showing chronic -pneumonia. The neutrophils were CR3-negative when characterized by flow cytometric analysis using anti-CD18 monoclonal antibody. Chemiluminescent response mediated by CR3 in neutrophils isolated from bronchoalveolar lavage fluid from BLAD-calves showed similar findings obtained from CR3-deficient neutrophils. Neutrophils from normal calves migrated to the scraped tissue, evaluated by skin window method. These findings confirmed the extravasating property of normal and CR3-deficient neutrophils into scraped tissues.Granulocyte transfusion as performed in an 8-month-old heifer with BLAD to monitor the changes in transfused CDl8-positive neutrophils and associated neutrophil chemilurninescent response in B2 integrin deficient host. The CD18-positive neutrophils were detected in blood from the BLAD heifer during the first 3 hr after 2.6xl0^9cells were infused by GT, and disappeared by 5 hr after GT.The CL response of neutrophils was increased 1.7 to 2.8-fold in the BLAD heifer during the first 3 hr after GT, thereafter CL response decreased gradually from 2 to 5 hr after GT.Bone marrow transplantation (BMT) was performed in a 9-month-old heifer with BLAD.Clinical and hematological findings, selected neutrophil function and CD18 expression of neutrophils in a B2 integrin-deficient heifer were examined. Twelve months after BMT, a small fluorescent region in the CD18-positive area of a flow cytometric profile was demonstrated and estimated to represent 0.3-0.5% of the CD18-positive neutrophils as measured by flow cytometric analysis following irnmunomagnetic separation. The animal's clinical condition appeared to improve and stabilize over our observation period of 28 months following BMT.Newly expressed CD18 seemed to play an important role in ameliorating the clinical signs of BLAD in this heifer. Less

进行了上述标题的研究，结果总结如下：将来自患有白细胞粘附缺陷的小母牛的补体受体3型（CR 3）缺陷型中性粒细胞的免疫球蛋白G介导的吞噬作用、超氧化物产生和细胞内钙（[Ca 2 +] i）信号传导的Fc受体与对照小母牛的Fc受体进行比较。IgG包被的酵母和聚集的牛IgG（Agg-IgG）诱导的CR 3缺陷型中性粒细胞的超氧化物生成的平均吞噬活性分别为对照中性粒细胞的10%和77.9%。在用Agg-IgG或伴刀豆球蛋白A刺激的CR 3缺陷型中性粒细胞中，[Ca^2 +]^i信号不足，平均峰值[Ca^2+]^i浓度为对照组的78%和42%。这些发现表明Fc受体介导的中性粒细胞功能与中性粒细胞表面CR 3的存在密切相关。 ...更多信息 间隙和刮下的组织与正常小牛的组织进行比较。在显示慢性肺炎的BLAD受影响小牛的支气管肺泡灌洗液中发现中性粒细胞。使用抗CD 18单克隆抗体进行流式细胞术分析时，中性粒细胞呈CR 3阴性。从BLAD小牛支气管肺泡灌洗液中分离的中性粒细胞中CR 3介导的化学发光反应显示出从CR 3缺陷中性粒细胞中获得的类似结果。皮肤窗法检测正常犊牛中性粒细胞向刮取组织的迁移。在患有BLAD的8月龄小母牛中进行粒细胞输注，以监测输注的CD 18阳性中性粒细胞和相关的中性粒细胞在B2整合素缺陷宿主中的瞬时应答的变化。在GT输注2.6x10^9个细胞后的前3小时内，在BLAD小母牛的血液中检测到CD 18阳性中性粒细胞，并在GT后5小时消失。在GT后的前3小时内，BLAD小母牛中中性粒细胞的CL反应增加了1.7至2.8倍，此后，CL反应从GT后2至5小时逐渐降低。在患有BLAD的9月龄小母牛中进行骨髓移植（BMT）。临床和血液学结果，在B2整联蛋白缺乏的小母牛中检测了选定的中性粒细胞功能和中性粒细胞的CD 18表达。BMT后12个月，在流式细胞术的CD 18阳性区域中显示了一个小的荧光区域，并估计代表0.3-0.5%的CD 18阳性中性粒细胞，如免疫磁性分离后通过流式细胞术分析所测量的。动物的临床状况出现改善和稳定在我们的观察期28个月后BMT。新表达的CD 18似乎在改善BLAD在这头母牛的临床症状中发挥了重要作用。少

项目成果

Higuchi,H.,et al.: "Relationship between the chemiluminescent response of borine neutrophils and chauges in 〔Ca^<2+>〕^i concentration." Can.J.Vet.Res.61(1). 57-61 (1997)

Higuchi, H., et al.：“硼中性粒细胞的化学发光反应与 [Ca^<2+>]^i 浓度的变化之间的关系。Can.J.Vet.Res.61(1)”。 (1997)

H.Nagahata: "Survival of transfused CD18-positive granulocytes and their chemiluminescent response in a Heifer with leukocyte adhesion deficiency." J.Vet.Med.Sci.60. 261-262 (1998)

H.Nagahata：“白细胞粘附缺陷的小母牛中输注的 CD18 阳性粒细胞的存活及其化学发光反应。”

H.Nagahata: "Fc receptor-mediated phagocytosis, superoxide production and calcium signaling of B2 integrin-deficient bovine neutrophils." Microbol.Immunol.41. 747-750 (1997)

H.Nagahata：“B2 整合素缺陷牛中性粒细胞的 Fc 受体介导的吞噬作用、超氧化物产生和钙信号传导。”

Nagahata, H.et al.: "Fcreceptor-mediated phagocytosis, superoxide production and calcium signaling of B_2 integrin-deficient neutrophils." Microbiol.lmmunol.41(9). 747-750 (1997)

Nagahata, H.et al.：“Fc 受体介导的 B_2 整合素缺陷型中性粒细胞的吞噬作用、超氧化物产生和钙信号传导。”

H.Nagahata: "Leukocyte emigration in normal calves and calves with leukocyte adhesion deficiency." J.Vet.Med.Sci.59. 1143-1147 (1997)

H.Nagahata：“正常犊牛和白细胞粘附缺陷犊牛的白细胞迁移。”