重组干扰素-γ同时增强单核细胞免疫调节特性和效应功能。

Concurrent enhancement of monocyte immunoregulatory properties and effector functions by recombinant interferon-gamma.

作者信息

Tweardy D J, Fujiwara H, Scillian J J, Ellner J J

出版信息

Cell Immunol. 1986 Jun;100(1):34-46. doi: 10.1016/0008-8749(86)90004-3.

Abstract

Interferon-gamma is a critical factor in the activation of several mononuclear phagocyte effector and immunoregulatory properties. However, it remains uncertain if IFN-gamma is capable of concurrent activation of both functions in the same cell population. Plastic adherent mononuclear cells (80-98% MN by cytochemical criteria) were cultivated in the absence or presence of recombinant interferon-gamma (rIFN-gamma, 0.1-100 U/ml) for 48 hr. MN surface DR antigen was assessed by flow cytometry (EPICS V) after staining with monoclonal antibodies OKIa1 or L243. Exposure to rIFN-gamma (100 U/ml) increased MN surface DR antigen (mean fluorescence intensity) by 80 +/- 20% (P less than 0.01) and 121 +/- 52% (P less than 0.001), respectively, compared to untreated cells. The increase in DR antigen was maximal at 100 U/ml, dependent on protein and RNA synthesis and blocked by agents that increase cAMP levels. IL-1 activity was determined in the mouse thymocyte assay; rIFN-gamma (100 U/ml) increased IL-1 activity in the supernatants of MN cultured in medium alone from 0.5 +/- 0.2 to 7.8 +/- 4.7 U/ml (P less than 0.05), and lipopolysaccharide-stimulated MN from 20.4 +/- 19.1 to 71.7 +/- 38.9 U/ml (P less than 0.05). Following rIFN-gamma exposure, MN stimulation of the AMLR was increased at 6 days (29,269 +/- 5224 vs 13,252 +/- 4938 cpm, P less than 0.01). Spontaneous cytotoxicity (SC) and antibody-dependent cell cytotoxicity (ADCC) were studied in a 51Cr release microculture assay using the human lymphoblastoid cell line CCRF-CEM as target. SC by MN increased linearly as a function of log[rIFN-gamma] for effector:target (E:T) ratios of 5:1 (r = 0.95, P less than 0.01) and 10:1 (r = 0.99, P less than 0.01). ADCC by MN increased following rIFN-gamma exposure (100 U/ml) at E:T ratios of 5:1 (22 +/- 13 to 31 +/- 4%, P less than 0.025) and 10:1 (31 +/- 4 to 38 +/- 4%, P less than 0.01). Thus, rIFN-gamma not only activates MN effector function, but has concurrent stimulatory effects on multiple MN properties critical to immunoregulation.

摘要

干扰素-γ是激活几种单核吞噬细胞效应和免疫调节特性的关键因素。然而，干扰素-γ是否能够在同一细胞群体中同时激活这两种功能仍不确定。将塑料贴壁单核细胞（根据细胞化学标准，单核细胞占80-98%）在不存在或存在重组干扰素-γ（rIFN-γ，0.1-100 U/ml）的情况下培养48小时。用单克隆抗体OKIa1或L243染色后，通过流式细胞术（EPICS V）评估单核细胞表面DR抗原。与未处理的细胞相比，暴露于rIFN-γ（100 U/ml）分别使单核细胞表面DR抗原（平均荧光强度）增加了80±20%（P<0.01）和121±52%（P<0.001）。DR抗原的增加在100 U/ml时最大，依赖于蛋白质和RNA合成，并被增加cAMP水平的试剂所阻断。在小鼠胸腺细胞试验中测定IL-1活性；rIFN-γ（100 U/ml）使单独在培养基中培养的单核细胞上清液中的IL-1活性从0.5±0.2增加到7.8±4.7 U/ml（P<0.05），使脂多糖刺激的单核细胞中的IL-1活性从20.4±19.1增加到71.7±38.9 U/ml（P<0.05）。在暴露于rIFN-γ后，第6天单核细胞对自体混合淋巴细胞反应（AMLR）的刺激增加（29269±5224对13252±4938 cpm，P<0.01）。使用人淋巴母细胞系CCRF-CEM作为靶细胞，在51Cr释放微量培养试验中研究自发细胞毒性（SC）和抗体依赖性细胞毒性（ADCC）。对于效应细胞：靶细胞（E:T）比例为5:1（r = 0.95，P<0.01）和10:1（r = 0.99，P<0.01）时，单核细胞的SC随log[rIFN-γ]呈线性增加。在E:T比例为5:1（22±13%至31±4%，P<0.025）和10:1（31±4%至38±4%，P<0.01）时，暴露于rIFN-γ（100 U/ml）后，单核细胞的ADCC增加。因此，rIFN-γ不仅激活单核细胞效应功能，而且对免疫调节至关重要的多种单核细胞特性具有同时刺激作用。