Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY 14642, USA.
J Virol Methods. 2012 May;181(2):170-6. doi: 10.1016/j.jviromet.2012.02.005. Epub 2012 Feb 23.
Activated platelets form transient aggregates with monocytes in circulation and have a half-life of approximately 30-60 min. These complexes are increased in various inflammatory conditions and are an early marker of myocardial infarction. HIV-1 infection is associated with chronic inflammation, and increased CD16⁺ inflammatory monocytes have been observed in these individuals, probably as a result of increased interaction with platelets. However, narrow detection period and platelet activation during sample processing pose significant problems in detecting platelet-monocyte complexes (PMCs). A method was standardized addressing these difficulties, to enumerate PMCs involving CD16⁺ or CD16⁻ monocytes in whole blood using flow cytometry. Blood collected from healthy individuals was treated with either collagen (for platelet activation) or LPS (for monocyte activation) and subsequently used to study effect of these treatments on PMC formation. This method was also validated for the ex vivo quantitation of PMCs in blood obtained from persons infected with HIV. The in vitro results demonstrated that platelet activation, but not monocyte activation, resulted in significant increase in PMC formation. There was a significant increase in CD16⁺ PMCs and platelet activation, in samples obtained from persons infected with HIV as compared to those without HIV infection. Furthermore, PMC percentages correlated positively with platelet activation. These findings improve the ability to detect PMCs and shed light on HIV pathogenesis.
活化的血小板与循环中的单核细胞形成短暂的聚集物，半衰期约为 30-60 分钟。这些复合物在各种炎症条件下增加，是心肌梗死的早期标志物。HIV-1 感染与慢性炎症有关，在这些个体中观察到 CD16⁺炎症性单核细胞增加，可能是由于与血小板的相互作用增加所致。然而，在检测血小板-单核细胞复合物（PMCs）时，狭窄的检测时间和血小板活化期间的样本处理带来了重大问题。已经标准化了一种方法，以使用流式细胞术在全血中计数涉及 CD16⁺或 CD16⁻单核细胞的 PMCs。从健康个体采集的血液用胶原（用于血小板活化）或 LPS（用于单核细胞活化）处理，然后用于研究这些处理对 PMC 形成的影响。该方法还经过验证，可用于定量检测从 HIV 感染者血液中获得的 PMCs。体外结果表明，血小板活化而不是单核细胞活化导致 PMC 形成显著增加。与未感染 HIV 的人相比，从感染 HIV 的人获得的样本中 CD16⁺ PMCs 和血小板活化显著增加。此外，PMC 百分比与血小板活化呈正相关。这些发现提高了检测 PMCs 的能力，并揭示了 HIV 发病机制。
