Srivastava A, Pearson H, Bryant J, Favaloro E, Coulits N, Jindra J, Wylie B
Haematology Department, Westmead Hospital, Sydney, NSW, Australia.
Vox Sang. 1993;65(2):146-50. doi: 10.1111/j.1423-0410.1993.tb02132.x.
With the aim of reducing the damage to platelets while effectively removing class I HLA antigens from their surfaces, we developed a new method using acidified chloroquine diphosphate. Platelets were treated with a 0.2 M solution of chloroquine diphosphate (pH 4.0). More than 90% of the platelets remained viable after treatment. While a marked reduction in reactions of acidified chloroquine-treated platelets with multispecific HLA antisera was noted in comparison with phosphate-buffered-saline-(PBS)-treated platelets, reactions with platelet-specific antibodies were preserved. This was demonstrated by immunofluorescence tests and solid-phase and monoclonal antibody immobilization of platelet antigen assays. Aggregation responses, though reduced in comparison with PBS-treated platelets, were still preserved after acidified chloroquine treatment. Ultrastructural analysis did not show any significant difference from PBS-treated platelets. We conclude that treatment of platelets with acidified chloroquine diphosphate is a simple and effective method for removing class I HLA antigens from their surfaces with minimal damage to their structure and function.
为了在有效去除血小板表面I类HLA抗原的同时减少对血小板的损伤，我们开发了一种使用酸化二磷酸氯喹的新方法。用0.2M二磷酸氯喹溶液（pH 4.0）处理血小板。处理后超过90%的血小板仍保持活力。与磷酸盐缓冲盐水（PBS）处理的血小板相比，酸化氯喹处理的血小板与多特异性HLA抗血清的反应明显减少，而与血小板特异性抗体的反应得以保留。这通过免疫荧光试验以及血小板抗原测定的固相和单克隆抗体固定化得到证实。与PBS处理的血小板相比，虽然酸化氯喹处理后聚集反应有所降低，但仍得以保留。超微结构分析显示与PBS处理的血小板没有任何显著差异。我们得出结论，用酸化二磷酸氯喹处理血小板是一种简单有效的方法，可在对其结构和功能造成最小损伤的情况下从其表面去除I类HLA抗原。
