Yari Fatemeh, Ahmadzadeh Noushin, Azadpour Shima, Vaeli Shahram
Research Center, Iranian Blood Transfusion Organization, 14665-1157 Tehran, Iran.
Indian J Hematol Blood Transfus. 2012 Sep;28(3):152-6. doi: 10.1007/s12288-011-0120-0. Epub 2011 Oct 20.
The demand for standard platelet concentrates (PCs) has continued to increase in the recent years. Infusible platelet membranes (IPM) prepared from new or outdated human platelets have been developed as an alternative to standard PCs, with the additional advantage of long shelf life and increased viral safety. Reduction of HLA antigens on the IPM has been assigned as one of the probable advantages of this product. In re-examining this issue, we studied the existence of HLA class I on the surface of IPM microparticles. In comparison we also surveyed HLA expression on the surface of the naturally occurred platelet-derived microparticles (nPMPs) during 7 days storage. Intended for producing IPM, PCs obtained from Iranian blood transfusion organization were lysed; virally inactivated with wet heat in the presence of a heat stabilizer and then sonicated. IPMs were separated using centrifugation and liquid-stored in 4°C. The expression of HLA class I antigens was surveyed using flow cytometry technique. HLA molecules were present on the microparticles. Shedding of HLA antigens was demonstrated from the surface of the both liquid-stored IPM and nPMPs during storage. Storage of IPM in 4°C was accompanied with significant reduction of HLA molecules. It seemed that achievement of HLA-free IPM could be impossible unless chloroquine treated platelets were used to prepare these microvesicles.
近年来,对标准血小板浓缩物(PCs)的需求持续增长。由新鲜或过期人血小板制备的可输注血小板膜(IPM)已被开发出来作为标准PCs的替代品,具有保质期长和病毒安全性提高的额外优势。降低IPM上的HLA抗原被认为是该产品的可能优势之一。在重新审视这个问题时,我们研究了IPM微粒表面是否存在HLA I类分子。作为对比,我们还调查了天然存在的血小板衍生微粒(nPMPs)在储存7天期间表面的HLA表达情况。为了制备IPM,将从伊朗输血组织获得的PCs进行裂解;在热稳定剂存在下用湿热进行病毒灭活,然后进行超声处理。通过离心分离IPM,并在4°C下液体储存。使用流式细胞术技术检测HLA I类抗原的表达。微粒上存在HLA分子。在储存期间,液体储存的IPM和nPMPs表面均出现HLA抗原脱落。将IPM储存在4°C会伴随着HLA分子的显著减少。除非使用经氯喹处理的血小板来制备这些微泡,否则似乎不可能获得无HLA的IPM。