Santos-Argumedo L, Guerra-Infante F, Quesada-Pascual F, Estrada-Parra S
Department of Cell Biology, Centro de Investigacion y Estudios Avanzados, I.P.N., Mexico, D.F., Mexico.
Int J Lepr Other Mycobact Dis. 1995 Mar;63(1):56-61.
In this work we describe the purification and characterization of armadillo immunoglobulins. The IgM was precipitated using low-strength ionic solution and further purified by filtration through Sephadex G-200. The IgG was obtained in pure form by precipitation of serum with ammonium sulfate and DEAE-cellulose ion exchange chromatography. The purity of these immunoglobulins was evaluated by polyacrylamide gel electrophoresis. The results showed 28-kDa light chains and 55-kDa and 70-kDa heavy chains for IgG and IgM, respectively. The rabbit antibodies against these molecules were used to prepare fluorescein (FITC) and peroxidase conjugates. The FITC conjugate was used to quantify IgM-bearing lymphocytes. An average of 17% of peripheral blood lymphocytes were sIgM+ from 14 healthy animals. Additionally, in the same animals we quantified lymphocytes with the capacity to form rosettes with sheep red-blood cells; the average for this marker was 10%. Also, the production of crossreacting antibodies to BCG was evaluated in healthy and Mycobacterium leprae-inoculated animals using the peroxidase conjugates. All animals with active infection recognized BCG antigens.
在本研究中，我们描述了犰狳免疫球蛋白的纯化和特性。使用低强度离子溶液沉淀IgM，并通过Sephadex G - 200过滤进一步纯化。通过用硫酸铵沉淀血清和DEAE - 纤维素离子交换色谱法获得纯形式的IgG。通过聚丙烯酰胺凝胶电泳评估这些免疫球蛋白的纯度。结果显示，IgG的轻链为28 kDa，重链分别为55 kDa和70 kDa；IgM的轻链为28 kDa，重链为70 kDa。针对这些分子的兔抗体用于制备荧光素（FITC）和过氧化物酶缀合物。FITC缀合物用于定量携带IgM的淋巴细胞。14只健康动物外周血淋巴细胞中平均有17%为sIgM +。此外，在相同的动物中，我们定量了具有与绵羊红细胞形成玫瑰花结能力的淋巴细胞；该标志物的平均值为10%。此外，使用过氧化物酶缀合物在健康动物和接种麻风分枝杆菌的动物中评估了对卡介苗交叉反应抗体的产生。所有活动性感染动物均识别卡介苗抗原。
