Cox Karina, North Margaret, Burke Michael, Singhal Hemant, Renton Sophie, Aqel Nayef, Islam Sabita, Knight Stella C
Antigen Presentation Research Group, Faculty of Medicine, Imperial College London, Northwick Park and St. Marks Campus, Harrow, Middlesex, United Kingdom.
J Leukoc Biol. 2005 Nov;78(5):1142-52. doi: 10.1189/jlb.1103532.
Plasmacytoid dendritic cells (PDC) constitute a distinct subset of DC found in human peripheral lymph nodes (LN), but little is known about their function. Cell suspensions were prepared from tumor draining LN (n=20) and control LN (n=11) of women undergoing surgical resection for primary breast cancer and elective surgery for benign conditions, respectively. Using four-color flow cytometry, human leukocyte antigen-DR+ DC subsets were identified phenotypically. The proportions and numbers of cells innately producing interleukin (IL)-4, IL-10, IL-12, and interferon-gamma (IFN-gamma) were also measured from intracellular accumulation of cytokine after blocking with monensin. All flow cytometry data were collected without compensation and were compensated off-line using the Winlist algorithm (Verity software). This package also provided the subtraction program to calculate percentage positive cells and intensity of staining. PDC (CD11c-, CD123+) expressed more cytokines than did myeloid DC (CD11c+) or CD1a+ putative "migratory" DC (P<0.001). LN PDC from patients with a good prognosis (px; n=11) demonstrated a relative increase in IL-12 and IFN-gamma expression (median IL-10:IL-12 ratio=0.78 and median IL-4:IFN-gamma ratio=0.7), and PDC from LN draining poor px cancer (n=9) showed a relative increase in IL-10 and IL-4 expression (median IL-10:IL-12 ratio=1.31 and median IL-4:IFN-gamma ratio=2.6). The difference in IL-4:IFN-gamma expression between good and poor px cancer groups was significant (P<0.05). Thus, PDC innately producing cytokines were identified in cell suspensions from human LN, and the character of PDC cytokine secretion may differ between two breast cancer prognostic groups. We speculate that a shift towards PDC IL-10 and IL-4 expression could promote tumor tolerance in LN draining poor px breast cancer.
浆细胞样树突状细胞(PDC)是存在于人类外周淋巴结(LN)中的一种独特的树突状细胞亚群,但其功能鲜为人知。分别从接受原发性乳腺癌手术切除的女性的肿瘤引流淋巴结(n = 20)和接受良性疾病择期手术的女性的对照淋巴结(n = 11)中制备细胞悬液。使用四色流式细胞术,从表型上鉴定人白细胞抗原-DR +树突状细胞亚群。在用莫能菌素阻断后,还通过细胞内细胞因子积累来测量天然产生白细胞介素(IL)-4、IL-10、IL-12和干扰素-γ(IFN-γ)的细胞比例和数量。所有流式细胞术数据均在未进行补偿的情况下收集,并使用Winlist算法(Verity软件)进行离线补偿。该软件包还提供了减法程序来计算阳性细胞百分比和染色强度。PDC(CD11c-,CD123 +)比髓样树突状细胞(CD11c +)或CD1a +推定的“迁移性”树突状细胞表达更多的细胞因子(P <0.001)。预后良好(px;n = 11)患者的淋巴结PDC表现出IL-12和IFN-γ表达相对增加(IL-10:IL-12比值中位数= 0.78,IL-4:IFN-γ比值中位数= 0.7),而引流预后不良px癌症(n = 9)的淋巴结中的PDC表现出IL-10和IL-4表达相对增加(IL-10:IL-12比值中位数= 1.31,IL-4:IFN-γ比值中位数= 2.6)。预后良好和预后不良px癌症组之间IL-4:IFN-γ表达的差异具有统计学意义(P <0.05)。因此,在人淋巴结的细胞悬液中鉴定出了天然产生细胞因子的PDC,并且在两个乳腺癌预后组之间,PDC细胞因子分泌的特征可能有所不同。我们推测,向PDC的IL-10和IL-4表达转变可能会促进引流预后不良px乳腺癌的淋巴结中的肿瘤耐受性。