Garbuglia A R, Salvi R, Di Caro A, Cappiello G, Montella F, Di Sora F, Recchia O, Lauria F, Benedetto A
Centre for Virology, S. Camillo Hospital, Rome, Italy.
AIDS. 1996 Jan;10(1):17-21. doi: 10.1097/00002030-199601000-00003.
We investigated a selected group of 11 non-progressor, HIV-infected individuals 20 months prior to this study and found that they all had undetectable levels of viral RNA expression in their peripheral blood lymphocytes (PBL). Phorbol 12-myristate 13-acetate (PMA) and phytohaemagglutinin (PHA) stimulation of PBL produced easily detectable amounts of HIV RNA in only two out of five of these patients. Here we report the results of the virological and clinical follow-up of nine non-progressors from this group. We verified the stability of their non-progressive status and attempted to correlate it to specific virological markers.
Proviral DNA in lymphocytes was tested by semi-quantitative polymerase chain reaction (PCR). Detection of unspliced (US) and multiple spliced (MS) HIV RNA species in unstimulated and stimulated lymphocytes was performed by reverse transcriptase-PCR (RT-PCR). The amount of p24 antigen released into the media of lymphocyte cultures was measured using a standard procedure. Lymphocyte populations were depleted of CD8 cells by immunomagnetic purging.
Follow-up of nine of these subjects showed that the patients who previously showed viral RNA activation following lymphocyte stimulation in vitro, developed a clinical and immunological progression characterized by CD4 count decline and lymphadenopathy. In contrast, all the other subjects maintained progression-free status throughout the follow-up period, with no detectable levels of HIV RNA in the PBL. Notably, this group of subjects showed no activation of viral RNA expression following stimulation of either undepleted or CD8-depleted lymphocytes in vitro.
The group of non-progressors studied was found to be heterogeneous regarding the stability of the non-progressive status during the follow-up period. Our results suggest that the activation of HIV RNA expression following PMA-PHA treatment of lymphocytes in vitro is an early marker for future progression of the disease.
在本研究开展的20个月前，我们对一组经挑选的11名未进展的HIV感染者进行了调查，发现他们外周血淋巴细胞（PBL）中的病毒RNA表达水平均无法检测到。在这5名患者中，仅2名患者的PBL经佛波酯12 - 肉豆蔻酸酯13 - 乙酸酯（PMA）和植物血凝素（PHA）刺激后产生了易于检测到的HIV RNA量。在此，我们报告该组9名未进展者的病毒学和临床随访结果。我们验证了他们未进展状态的稳定性，并试图将其与特定的病毒学标志物相关联。
通过半定量聚合酶链反应（PCR）检测淋巴细胞中的前病毒DNA。通过逆转录 - 聚合酶链反应（RT - PCR）检测未刺激和刺激后的淋巴细胞中未剪接（US）和多剪接（MS）HIV RNA种类。使用标准程序测量释放到淋巴细胞培养上清中的p24抗原量。通过免疫磁珠清除法去除淋巴细胞群体中的CD8细胞。
对其中9名受试者的随访表明，先前在体外淋巴细胞刺激后出现病毒RNA激活的患者，出现了以CD4细胞计数下降和淋巴结病为特征的临床和免疫进展。相比之下，所有其他受试者在整个随访期间均保持无进展状态，PBL中未检测到HIV RNA水平。值得注意的是，该组受试者在体外未去除或去除CD8的淋巴细胞刺激后均未出现病毒RNA表达激活。
研究发现，所研究的未进展者群体在随访期间未进展状态的稳定性方面存在异质性。我们的结果表明，体外PMA - PHA处理淋巴细胞后HIV RNA表达的激活是该疾病未来进展的早期标志物。
