Abstract：
Innate and adaptive immune responses at mucosal surfaces play a role in protection against most infectious diseases. However, the relative importance either of mucosal versus systemic, or of cellular versus humoral immunity in protection against such infections remains unclear. We aimed to determine the relative percentages and reproducibility of detection of five major T lymphocyte phenotypes in stimulated whole mouth fluid (SWMF); to compare matched mucosal and blood phenotypes; to evaluate the consistency of phenotypes in SWMF over time; and to determine any associations with age or gender. Peripheral blood and SWMF samples were collected from 194 participants and sequential concomitant samples were collected from 27 of those and from 12 subjects living with HIV. CD3, CD4, CD8, Th1 and Th2 T lymphocyte phenotypes were determined by FACS. All the five T lymphocyte phenotypes were detected consistently by FACS in PBMC and SWMF with experimental replicates (N = 10; PBMC CV: 3-30%; SWMF CV: 12-36%). In longitudinal samples detection rates were reproducible in both fluids but variations were higher in SWMF (CV: 23-79.6%) than PBMC (CV: 9.7-75%). Statistically significant correlations of the percentages of all the T lymphocyte phenotypes except CD8 was seen between the two fluids. In PBMCs a negative correlation with age was found with CD3, CD4 and CD8 phenotypes, whilst a positive correlation was found in both SWMF and PBMC with the Th2 phenotype. CD3, CD4 and CD8 phenotypes in SWMF and PBMCs from an HIV-positive cohort were not significantly correlated in contrast with the HIV-negative controls. Our study provides a robust FACS protocol for the detection of the five major T lymphocyte phenotypes in SWMF which should prove useful for research with other mucosal fluids.
摘要：
粘膜表面的固有和适应性免疫应答在预防大多数传染病方面发挥作用。然而,粘膜与全身的相对重要性，或细胞对体液免疫在预防此类感染中的作用尚不清楚。我们旨在确定刺激的全口液（SWMF）中五种主要T淋巴细胞表型检测的相对百分比和可重复性；比较匹配的粘膜和血液表型；评估SWMF表型随时间的一致性；并确定与年龄或性别的任何关联。从194名参与者收集外周血和SWMF样品，并从其中27名和12名HIV感染者中收集顺序伴随样品。FACS测定CD3、CD4、CD8、Th1和Th2T淋巴细胞表型。通过FACS在PBMC和SWMF中一致地检测到所有五种T淋巴细胞表型，并具有实验重复（N=10；PBMCCV：3-30％；SWMFCV：12-36％）。在两种流体中，纵向样品的检出率均可重复，但SWMF（CV：23-79.6％）的变化高于PBMC（CV：9.7-75％）。在两种液体之间观察到除CD8以外的所有T淋巴细胞表型百分比的统计学显着相关性。在PBMC中，CD3，CD4和CD8表型与年龄呈负相关，而在SWMF和PBMC中发现与Th2表型呈正相关。与HIV阴性对照相比，来自HIV阳性队列的SWMF和PBMC中的CD3，CD4和CD8表型没有显着相关。我们的研究提供了一个强大的FACS协议，用于检测SWMF中的五种主要T淋巴细胞表型，这对于其他粘膜液的研究应该是有用的。
