Tamura Y, Lai P K, Bradley W G, Konno K, Tanaka A, Nonoyama M
Laboratory of Virology, Tampa Bay Research Institute, St. Petersburg, FL 33716.
Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2249-53. doi: 10.1073/pnas.88.6.2249.
We showed that an extract (PC6) from cones of Pinus parviflora Sieb et Zucc induced the human T-cell line CEM to produce a pepsin-sensitive soluble factor(s) that could inhibit the replication of the type 1 human immunodeficiency virus (HIV-1) in CEM T cells, in U-937 histocytes, in THP-1 monocytes, and in mitogen-activated human tonsillar mononuclear cells. Indirect immunofluorescence staining and polymerase chain reaction analysis of the PC6-induced CEM cells revealed the absence of known lymphokines/cytokines except granulocyte/macrophage colony-stimulating factor (GM-CSF), interleukin 3 (IL-3), transforming growth factor beta 1 (TGF-beta 1), and tumor necrosis factor alpha (TNF-alpha). However, functional studies with recombinant IL-3, TNF-alpha, and TGF-beta 1 showed that these three factors did not inhibit HIV-1 replication in CEM cells. Neutralization of the PC6-induced HIV-1-inhibiting factor(s) with commercially available neutralizing antibodies to GM-CSF and TNF-alpha also did not abrogate the anti-HIV-1 impact. Thus, the anti-HIV-1 factor induced by PC6 may be novel. Molecular sieve separation showed that the anti-HIV-1 factor(s) is smaller than 30 kDa. Affinity chromatography using a DEAE-cellulose column enriched the factor that inhibited HIV-1.
我们发现，五针松球果提取物（PC6）可诱导人T细胞系CEM产生一种对胃蛋白酶敏感的可溶性因子，该因子能够抑制1型人类免疫缺陷病毒（HIV-1）在CEM T细胞、U-937组织细胞、THP-1单核细胞以及丝裂原激活的人扁桃体单核细胞中的复制。对PC6诱导的CEM细胞进行间接免疫荧光染色和聚合酶链反应分析，结果显示除粒细胞/巨噬细胞集落刺激因子（GM-CSF）、白细胞介素3（IL-3）、转化生长因子β1（TGF-β1）和肿瘤坏死因子α（TNF-α）外，不存在已知的淋巴因子/细胞因子。然而，对重组IL-3、TNF-α和TGF-β1进行的功能研究表明，这三种因子均不能抑制CEM细胞中的HIV-1复制。用市售的针对GM-CSF和TNF-α的中和抗体中和PC6诱导的HIV-1抑制因子，也不能消除其抗HIV-1的作用。因此，PC6诱导产生的抗HIV-1因子可能是一种新的因子。分子筛分离显示，抗HIV-1因子的分子量小于30 kDa。使用DEAE-纤维素柱进行亲和层析，可富集抑制HIV-1的因子。
