Abstract

Cytidine monophosphate-Nacetylneuraminic acid (Neu5Ac) hydroxylase (CMAH) and glycoprotein, alpha1, 3-galactosyltransferase (GGTA1) double knockout (DKO) pig models were produced to reduce immune reaction for xenotransplantation. However, the role of Neu5Gc and α-Gal in pigs has not been fully elucidated and it is necessary to consider the after-effect of inactivation of GGTA1 and CMAH in pigs. Hematological profiles of DKO pigs were analyzed through complete blood count (CBC). Histology of liver and spleen of DKO were investigated, and lectin blotting and mass spectrometry (MS) were performed to explore glycosylation changes in red blood cell (RBC) membranes of DKO pigs. DKO pigs showed common clinical signs such as weakness (100%), dyspnea (90%) and constipation (65%). DKO pigs revealed a significant decrease in RBC, hemoglobin (HGB) and hematocrit (HGB), and an increase in white blood cell (WBC), lymphocyte (LYM), monocyte (MON), and erythrocyte mean corpuscular volume (MCV). DKO piglets showed swollen liver and spleen, and exhibited raised deposition of hemosiderin and severe bleeding. Lectin assay and MS proved variations in glycosylation on RBC membranes. GGTA1/CMAH DKO pigs developed pathological features which are similar to anemic symptoms, and the variations in glycosylation on RBC membranes of DKO pigs may be attributed to the pathologies observed.

摘要

胞苷单磷酸-Nacetylneuraminic 酸 (Neu5Ac) 羟化酶 ( CMAH ) 和糖蛋白、α1, 3-半乳糖基转移酶 ( GGTA1 ) 双敲除 (DKO) 猪模型被生产以减少异种移植的免疫反应。然而，Neu5Gc和α-Gal在猪体内的作用尚未完全阐明，需要考虑GGTA1和CMAH失活后遗症在猪中。通过全血细胞计数 (CBC) 分析 DKO 猪的血液学特征。对 DKO 猪的肝脏和脾脏进行了组织学研究，并进行了凝集素印迹和质谱分析 (MS)，以探索 DKO 猪红细胞 (RBC) 膜的糖基化变化。DKO 猪表现出常见的临床症状，例如虚弱 (100%)、呼吸困难 (90%) 和便秘 (65%)。DKO 猪的红细胞、血红蛋白 (HGB) 和血细胞比容 (HGB) 显着下降，白细胞 (WBC)、淋巴细胞 (LYM)、单核细胞 (MON) 和红细胞平均红细胞体积 (MCV) 增加。DKO仔猪肝脾肿大，含铁血黄素沉积增多，出血严重。凝集素测定和 MS 证实了 RBC 膜上糖基化的变化。GGTA1/CMAHDKO 猪出现了类似于贫血症状的病理特征，并且 DKO 猪红细胞膜上糖基化的变化可能归因于所观察到的病理。