One-step concentration of malarial parasite-infected red blood cells and removal of contaminating white blood cells
One-step concentration of malarial parasite-infected red blood cells and removal of contaminating white blood cells
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DOI：
10.1186/1475-2875-3-7
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发表时间：
2004-03-01
期刊：
影响因子：
3
通讯作者：
Kamei, K
中科院分区：
文献类型：
--
作者：
Trang, DTX;Huy, NT;Kamei, K
背景资料：分离浓缩的、活的、受疟疾寄生虫感染的红细胞（PRBCs）的制备物，其具有低的白色血细胞（WBC）污染，促进了对疟疾寄生虫的分子、生物化学和免疫学方面的研究。这目前通过两步法进行，包括通过Percoll或Nycodenz使用密度梯度离心浓缩PRBC，然后使用纤维素粉末柱或市售过滤装置去除宿主WBC。这些两步法可以帮助分离足够的PRBC，但它们是费力的。在这项研究中，一个简化的一步程序，利用抗磁性低自旋氧合血红蛋白和顺磁性血红素（血红素聚合物）之间的差异进行了描述。顺磁性聚合物沉积在寄生虫的食物泡中，允许使用磁分离来有效地和快速地浓缩PRBCs，同时去除污染的宿主WBCs.Methods：使用商业LD柱（MACS）的WBC的磁性去除被评估为一种新的方法用于浓缩和纯化PRBCs。为了比较这种方法与两种密度梯度离心法，使用Percoll或Nycodenz，我们分析了富集PRBCs和污染的主机白细胞的数量，以及在最终preparation.Results的疟疾寄生虫的活力：PRBCs的数量和疟疾寄生虫的活力之间的磁性和离心法分离的PRBCs是相似的。然而，90-99%的污染白细胞被删除从起始材料使用磁性柱，而白细胞含量没有改变使用Percoll或Nycodenz methods.Conclusion：使用市售的磁性LD柱是有效的，安全的，易于一步纯化PRBCs。这种简单的方法不会影响疟疾寄生虫的生存能力。
Background: Isolation of a concentrated, living preparation of malarial parasite-infected red blood cells (PRBCs) that have low contamination of white blood cells (WBCs) facilitates research on the molecular, biochemical and immunological aspects of malarial parasites. This is currently carried out by a two-step method, including the concentration of PRBCs using density gradient centrifugation through Percoll or Nycodenz, followed by the removal of host WBCs using a cellulose powder column or a commercially available filtration unit. These two-step methods can help isolate sufficient PRBCs, but they are laborious. In this study, a simplified one-step procedure that takes advantage of the difference between diamagnetic low-spin oxyhaemoglobin and paramagnetic haemozoin (haem polymer) was described. The paramagnetic polymer is deposited in the food vacuoles of the parasite, allowing the use of magnetic separation to efficiently and rapidly concentrate PRBCs while removing contaminating host WBCs.Methods: The magnetic removal of WBCs using a commercial LD column ( MACS) was evaluated as a new method for concentrating and purifying PRBCs. To compare this method with the two density gradient centrifugation methods using Percoll or Nycodenz, we analysed the quantities of enriched PRBCs and contaminating host WBCs as well as the viability of malarial parasites in the final preparations.Results: The quantity of PRBCs and the viability of malarial parasites in the isolated PRBCs were similar between magnetic and centrifugation methods. However, 90-99% of the contaminating WBCs were removed from the starting material using a magnetic column, whereas WBC content did not change using the Percoll or Nycodenz methods.Conclusion: The use of a commercially available magnetic LD column is effective, safe and easy for the one-step purification of PRBCs. This simple method does not affect the viability of malarial parasites.