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

研究结果总结如下：我们比较了补体受体3-型（CR3）缺陷的中性粒细胞与白细胞粘附缺乏症母牛的Fc受体介导的免疫球蛋白g介导的吞噬、超氧化物产生和细胞内钙（[Ca^2+]^i）信号传导。IgG包被酵母和聚集牛IgG （Agg-IgG）诱导的cr3缺陷中性粒细胞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%。受Agg-Igg或concanavin A刺激的cr3缺陷中性粒细胞的[Ca^2+]^i信号缺乏，平均峰值[Ca^2+]^i浓度分别为对照组的78%和42%。这些发现表明Fc受体介导的中性粒细胞功能密切依赖于中性粒细胞表面CR3的存在。与正常犊牛相比，B2整合素缺乏症（BLAD）犊牛白细胞向支气管肺泡内迁移。慢性肺炎患儿支气管肺泡灌洗液中发现中性粒细胞。用抗cd18单克隆抗体进行流式细胞分析，证实中性粒细胞为cr3阴性。从brd小牛支气管肺泡灌洗液中分离的中性粒细胞中由CR3介导的化学发光反应显示了从CR3缺失的中性粒细胞中获得的类似结果。正常犊牛中性粒细胞向刮伤组织迁移，采用皮肤窗法评估。这些发现证实了正常和cr3缺陷中性粒细胞向刮伤组织的外渗特性。在8个月大的BLAD小母牛中进行粒细胞输注，以监测输注cd18阳性中性粒细胞和相关中性粒细胞化学发光反应在B2整合素缺乏宿主中的变化。转染2.6 × 10 ^9细胞后3小时内，BLAD小母牛血液中检测到cd18阳性中性粒细胞，并在5小时内消失，中性粒细胞CL反应在转染后3小时内增加1.7 ~ 2.8倍，随后在2 ~ 5小时内逐渐下降。研究了B2整合素缺陷小母牛的临床和血液学表现、中性粒细胞选择功能和中性粒细胞CD18的表达。BMT后12个月，流式细胞术显示cd18阳性区域的小荧光区域，估计代表免疫磁分离后流式细胞术分析测量的cd18阳性中性粒细胞的0.3-0.5%。在BMT后的28个月的观察期间，动物的临床状况似乎有所改善和稳定。新表达的CD18似乎在改善该母牛BLAD的临床症状中起重要作用。少

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