The role of platelet microparticles in the production of antibodies from B lymphocytes against HLA-DR antigen in vitro

IF 0.4 Q4 PEDIATRICS
Zahra Khayati, F. Yari
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引用次数: 1

Abstract

Background: Platelets can activate B cells and stimulate them for the production of antibodies. Since platelet microparticles (PMPs) originate from platelets, they may have the same virtue. In the present study, the effect of PMPs was investigated on the production of human leukocyte antigen (HLA)-specific antibody from B cells in vitro. Materials and Methods: In this experimental study, HLA-DR antigen was solubilized from the immortalized B lymphocytes (Daudi cell line) and purified using the affinity chromatography. Antigen properties were determined by the ELISA technique. PMPs were isolated from platelet concentrate bags by centrifugation. Fresh blood products were prepared from the Innovation Center of Iranian Blood Transfusion Organization (IBTO) and B lymphocytes were purified by the MACS method. B cells were exposed with PMPs and HLA-DR antigens in the culture medium. On the third day of culture, the culture supernatant was examined in terms of antibody production using the ELISA test. The results were analyzed using paired sample T-test and P-value <0.05 was considered statistically significant. Results: The specificity of the purified HLA-DR antigen was confirmed using the anti-HLA-DR antibody and ELISA technique in the presence of appropriate controls. The results showed that PMPs could stimulate the production of antibodies from B cells. The difference between the case and control was significant (P-value=0.001). Although total immunoglobulin (IgG) was higher in HLA-DR-treated wells, HLA-DR-specific antibodies were not identified by ELISA technique.  Conclusion: PMPs have the capability to induce IgG antibodies from B cells. In order to ensure the production of specific antibodies, further testing is required with high sensitivity.
血小板微粒在B淋巴细胞体外产生抗HLA-DR抗原抗体中的作用
背景:血小板可以激活B细胞并刺激其产生抗体。由于血小板微粒(pmp)起源于血小板,它们可能具有相同的优点。本研究探讨了pmp对B细胞产生人白细胞抗原(HLA)特异性抗体的影响。材料和方法:从永生化B淋巴细胞(Daudi细胞系)中溶解HLA-DR抗原,采用亲和层析纯化。采用酶联免疫吸附法测定抗原性质。用离心法从血小板浓缩袋中分离出pmp。从伊朗输血组织创新中心(IBTO)制备新鲜血液制品,用MACS法纯化B淋巴细胞。将B细胞暴露于pmp和HLA-DR抗原的培养基中。培养第3天,用ELISA法检测培养上清液的抗体产生情况。结果采用配对样本t检验,p值<0.05为差异有统计学意义。结果:在存在适当对照的情况下,用抗HLA-DR抗体和ELISA技术证实纯化的HLA-DR抗原的特异性。结果表明,pmp可以刺激B细胞产生抗体。病例与对照组的差异有统计学意义(p值=0.001)。虽然总免疫球蛋白(IgG)在hla - dr处理的井中较高,但ELISA技术未检测到hla - dr特异性抗体。结论:pmp具有诱导B细胞产生IgG抗体的能力。为了确保产生特异性抗体,需要进一步进行高灵敏度的检测。
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来源期刊
CiteScore
0.80
自引率
33.30%
发文量
33
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