Holt P G, Kees U R, Shon-Hegrad M A, Rose A, Ford J, Bilyk N, Bowman R, Robinson B W
Clinical Immunology Research Unit, Princess Margaret Hospital, Subiaco, West Australia.
Immunology. 1988 Aug;64(4):649-54.
This study evaluates the frequency and functions of immunocompetent T cells at the clonal level in solid human lung tissue versus peripheral blood. Enzymatic digestion of slices of histologically normal human lung yielded 18-42 x 10(6) viable mononuclear cells per gram wet weight tissue, of which 60-72% were lymphocytes; based upon these recoveries and the known weight of adult lung, the (median) lung parenchymal lymphocyte population can be estimated as 6 x 10(9), being of the same order as the blood pool and 15-30-fold that recoverable by broncho-alveolar lavage. Flow cytometric analysis indicated that the bulk of these lymphocytes was OKT3+/T11+ (CD3/CD2) T cells. Purified blood and lung T cells from each subject were cultured at limiting dilution in the presence of PHA, irradiated feeder cells and recombinant human IL-2. The mean frequency estimates for PHA-responsive T cells in these populations were 1 in 1.23 (81%) and 1 in 3.22 (31%) for blood and lung, respectively. This difference was seen for T cells from each donor and was highly significant by paired t-test (P less than 0.002). Analysis of surface phenotypes and functions of individual blood and T-cell clones indicated comparable frequencies for OKT4 (CD4) and OKT8 (CD8) expression, TNF production and mitogen-induced cytotoxicity. However, a striking inverse relationship was observed between the overall frequency of IL-2-producing clones (79% for blood versus 47% for lung) and interferon-gamma (IFN-gamma)-producing clones (46% versus 87%). These differences were found for each subject, and both were highly significant (P less than 0.001) by paired t-test. The available literature suggests that the majority of these lung T cells represent transient immigrants derived from the blood. Accordingly, the functional differences we have observed suggest either selective trapping within the lung vascular bed of peripheral blood T cells of certain functional phenotypes or alternatively selection/modulation of T cells by lung-derived factors during their transit through the tissue.
本研究在克隆水平评估人实体肺组织与外周血中免疫活性T细胞的频率及功能。对组织学正常的人肺切片进行酶消化,每克湿重组织可获得18 - 42×10⁶个活的单核细胞,其中60 - 72%为淋巴细胞;根据这些回收率及已知的成人肺重量,(中位数)肺实质淋巴细胞群体估计为6×10⁹,与血池数量相当,是通过支气管肺泡灌洗可回收数量的15 - 30倍。流式细胞术分析表明,这些淋巴细胞大部分为OKT3⁺/T11⁺(CD3/CD2)T细胞。从每个受试者中纯化的血液和肺T细胞在PHA、照射过的饲养细胞和重组人IL - 2存在的情况下进行有限稀释培养。这些群体中对PHA反应性T细胞的平均频率估计分别为血液中的1/1.23(81%)和肺中的1/3.22(31%)。每个供体的T细胞均出现这种差异,经配对t检验差异极显著(P<0.002)。对单个血液和T细胞克隆的表面表型及功能分析表明,OKT4(CD4)和OKT8(CD8)表达、TNF产生及丝裂原诱导的细胞毒性的频率相当。然而,观察到产生IL - 2的克隆的总体频率(血液中为79%,肺中为47%)与产生干扰素 - γ(IFN - γ)的克隆的总体频率(46%对87%)之间存在显著的负相关关系。每个受试者均发现这些差异,经配对t检验两者均极显著(P<0.001)。现有文献表明,这些肺T细胞大多数代表源自血液的短暂性移民。因此,我们观察到的功能差异表明,要么是外周血中具有某些功能表型的T细胞在肺血管床中被选择性截留,要么是T细胞在通过组织的过程中被肺源性因子进行选择/调节。