DIRECT ACTIVITY OF HUMAN T-LYMPHOCYTES AND NATURAL-KILLER-CELLS AGAINST CRYPTOCOCCUS-NEOFORMANS
DIRECT ACTIVITY OF HUMAN T-LYMPHOCYTES AND NATURAL-KILLER-CELLS AGAINST CRYPTOCOCCUS-NEOFORMANS
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DOI：
10.1128/iai.62.1.194-202.1994
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发表时间：
1994-01-01
影响因子：
3.1
通讯作者：
SMAIL, EH
中科院分区：
文献类型：
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作者：
LEVITZ, SM;DUPONT, MP;SMAIL, EH
淋巴细胞构成宿主防御隐球菌病的关键组成部分，以前，我们证明了人淋巴细胞培养白细胞介素-2形成共轭物，并直接抑制，新生隐球菌的生长。在这里，我们探讨了新鲜分离的，高度纯化的人外周血淋巴细胞群体的抗隐球菌活性。淋巴细胞与包囊C.新生儿24小时，之后裂解淋巴细胞，制备稀释液并涂布平板，并计数CFU。通过比较有淋巴细胞和无淋巴细胞的威尔斯孔中的生长来确定抑真菌作用。尼龙羊毛-非粘附性外周血单核细胞（NWNA PBMC）具有高度抑真菌性，即使T细胞或自然杀伤（NK）细胞通过淘洗被耗尽。通过用绵羊红细胞重置NWNA PBMC获得的T细胞和NK细胞的混合群体完全抑制隐球菌的生长，而nonrosetting细胞几乎没有抑真菌活性。通过阳性免疫选择分离的CD 4（+）、CD 8（+）和CD 16/56（+）淋巴细胞具有强的生长抑制活性。相反，纯化的B细胞没有活性。甚至在没有调理素的情况下也观察到抑真菌作用。当用胰蛋白酶或菠萝蛋白酶处理NWNA PBMC上的表面受体时，抗真菌活性显著降低。来自刺激的淋巴细胞或浓缩的淋巴细胞超声处理物的上清液没有活性。淋巴细胞介导的抑菌作用在两种不同的C.新人类CD 4（+）、CD 8（+）和CD 16/56（+）淋巴细胞与C.在Nomarski微分干涉相差显微镜和视频显微镜下观察到的新形式。这些数据表明，新鲜分离的外周血T细胞和NK细胞具有结合并直接抑制C.新人类
Lymphocytes constitute a critical component of host defenses against cryptococcosis, Previously, we demonstrated that human lymphocytes cultured with interleukin-2 formed conjugates with, and directly inhibited the growth of, Cryptococcus neoformans. Here, we explore the anticryptococcal activity of freshly isolated, highly purified populations of human peripheral blood lymphocytes. Lymphocytes were incubated with encapsulated C. neoformans for 24 h, after which the lymphocytes were lysed, dilutions and spread plates were made, and CFU were counted. Fungistasis was determined by comparing growth in wells with and without lymphocytes. Nylon wool-nonadherent peripheral blood mononuclear cells (NWNA PBMC) were highly fungistatic, even if either T cells or natural killer (NK) cells were depleted by panning. A mixed population of T cells and NK cells, obtained by resetting NWNA PBMC with sheep erythrocytes, completely inhibited cryptococcal growth, whereas the nonrosetting cells had little fungistatic activity. CD4(+), CD8(+), and CD16/56(+) lymphocytes, isolated by positive immunoselection, had potent growth-inhibitory activity. In contrast, purified B cells had no activity. Fungistasis was seen even in the absence of opsonins. Antifungal activity was markedly diminished when surface receptors on NWNA PBMC were cleaved by treatment with trypsin or bromelain, Supernatants from stimulated lymphocytes or concentrated lymphocyte sonicates were not active. Lymphocyte-mediated fungistasis was seen with two different strains of C. neoformans. CD4(+), CD8(+), and CD16/56(+) lymphocytes formed conjugates with C. neoformans, as observed under Nomarski differential interference contrast microscopy and videomicroscopy. These data demonstrate that freshly isolated peripheral blood T cells and NK cells have the capacity to bind and directly inhibit the growth of C. neoformans.