Pitrak D L, Tsai H C, Mullane K M, Sutton S H, Stevens P
Department of Medicine, University of Illinois College of Medicine at Chicago, West Side VA Medical Center, 60612, USA.
J Clin Invest. 1996 Dec 15;98(12):2714-9. doi: 10.1172/JCI119096.
Neutrophil (PMNL) function defects occur as a consequence of HIV infection. This study examined PMNL apoptosis in patients with the acquired immunodeficiency syndrome (AIDS) to determine if accelerated apoptosis contributes to impaired function. PMNL were isolated from 10 HIV-infected patients with CD4+ lymphocyte counts < 200/mm3 without signs of active infection and 7 healthy volunteers. PMNL were stained with acridine orange and ethidium bromide after 0, 3, 6, and 18 h in culture, and examined for the morphologic changes of apoptosis and viability by fluorescent microscopy. Apoptosis was also demonstrated by electron microscopy, flow cytometry, and DNA gel electrophoresis. Apoptosis was minimal at 0 h, but PMNL from AIDS patients exhibited significantly greater apoptosis than controls at 3 h (22.5+/-11.5 vs. 8.9+/-6.9%, P = 0.015), 6 h (38.1+/-14.2 vs. 18.1+/-4.5%, P = 0.003), and 18 h (71.3+/-19.0 vs. 38.8+/-16.7%, P = 0.002). Viabilities were > or = 88.0% for both groups from 0-6 h, but by 18 h viability was significantly decreased for the HIV group (58.8+/-12.4 vs. 83.5+/-10.4%, P = 0.001) due to an increase in non-viable apoptotic cells. Incubation with serum from AIDS patients had no effect on control PMNL, and incubation with control serum did not reduce the rate of apoptosis of PMNL from AIDS patients. Incubation with granulocyte colony-stimulating factor (G-CSF) in vitro significantly decreased apoptosis for PMNL from AIDS patients. PMNL from patients with AIDS exhibit markedly accelerated apoptosis ex vivo. In vivo, apoptosis and functional impairment of PMNL may contribute to the risk of secondary infections, and cytokine therapy may be of potential clinical benefit in this circumstance.
中性粒细胞（PMNL）功能缺陷是HIV感染的结果。本研究检测了获得性免疫缺陷综合征（AIDS）患者的PMNL凋亡情况，以确定加速凋亡是否导致功能受损。从10例CD4 +淋巴细胞计数<200/mm3且无活动性感染迹象的HIV感染患者和7名健康志愿者中分离出PMNL。培养0、3、6和18小时后，用吖啶橙和溴化乙锭对PMNL进行染色，并通过荧光显微镜检查凋亡和活力的形态学变化。还用电子显微镜、流式细胞术和DNA凝胶电泳证实了凋亡。0小时时凋亡极少，但AIDS患者的PMNL在3小时（22.5±11.5对8.9±6.9%，P = 0.015）、6小时（38.1±14.2对18.1±4.5%，P = 0.003）和18小时（71.3±19.0对38.8±16.7%，P = 0.002）时的凋亡明显高于对照组。两组在0至6小时的活力均≥88.0%，但到18小时时，HIV组的活力因非存活凋亡细胞增加而显著降低（58.8±12.4对83.5±10.4%，P = 0.001）。与AIDS患者的血清孵育对对照PMNL无影响，与对照血清孵育也未降低AIDS患者PMNL的凋亡率。体外与粒细胞集落刺激因子（G-CSF）孵育可显著降低AIDS患者PMNL的凋亡。AIDS患者的PMNL在体外表现出明显加速的凋亡。在体内，PMNL的凋亡和功能损害可能导致继发感染的风险，在这种情况下细胞因子治疗可能具有潜在的临床益处。
