Papadavid E, Economidou J, Psarra A, Kapsimali V, Mantzana V, Antoniou C, Limas K, Stratigos A, Stavrianeas N, Avgerinou G, Katsambas A
Department of Dermatology and Venereology of the University of Athens, A.Syngros Hospital, Athens, Greece.
Br J Dermatol. 2003 Apr;148(4):709-18. doi: 10.1046/j.1365-2133.2003.05224.x.
There is evidence that a T-helper (Th) 2 cytokine pattern dominates in the peripheral blood as well as in tissue of patients with Sézary syndrome (SS), and that the malignant clone is of Th2 phenotype. However, there are conflicting studies on the cytokine pattern in the peripheral blood in different stages of cutaneous T-cell lymphoma (CTCL).
To examine, by means of flow cytometry (FC), the Th1/Th2 cytokine profile [cytoplasmic interferon (IFN)-gamma/interleukin (IL)-4] in peripheral blood T cells from patients with mycosis fungoides (MF) and SS, the most common forms of CTCL, and to correlate their expression with clinical stage, clonality and T-cell immunophenotype changes in order to evaluate their relevance in CTCL progression.
We investigated by FC the percentage of CD3+ T cells expressing cytoplasmic IFN-gamma and IL-4 after stimulation in blood specimens of 43 CTCL patients (32 stage I-II and 11 stage III-IV), eight of whom were erythrodermic. Next, we compared cytoplasmic IFN-gamma and IL-4 expression between patients of different stages and controls, and correlated our findings to T-cell receptor (TCR)-gamma gene rearrangement, used as a marker of clonality, and changes in T-cell immunophenotype (CD4+, CD8+, CD4+/CD7-, CD4+/CD25+) and natural killer cells. Polymerase chain reaction amplification of the TCR-gamma gene was performed in 41 blood and 26 skin specimens. We also examined the cytokine expression pattern in patients with erythrodermic MF and SS.
A significantly higher frequency of CD3+/IL-4+ T cells was found in late (III-IV) compared with early (I-II) CTCL patients (P = 0.002) or controls (P < 0.001). There were significant positive correlations between the percentages of CD3+/IL-4+ and the percentages of CD3+/CD4+ T cells (r = 0.385, P = 0.05), CD4+/CD7- T cells (r = 0.335, P < 0.05) and CD4+/CD25+ T cells (r = 0.433, P = 0.01); there was a negative correlation between the percentages of CD3+/IL-4+ and CD3+/CD8+ T cells (r = -0.463, P = 0.005) and a positive correlation between the percentages of CD3+/IFN-gamma+ and CD3+/CD8+ T cells (r = 0.368, P = 0.02). Increased percentages of CD3+/IL-4+, CD3+/CD4+ and CD4+/CD7- T lymphocytes were associated with the presence of clonality (P = 0.025, P < 0.001 and P = 0.0031, respectively). All independent variables showed a statistically significant difference between SS and erythrodermic MF patients, or controls, apart from cytoplasmic IL-4, which was high both in erythrodermic MF and SS patients compared with controls (P = 0.003 and P = 0.008, respectively). In multiple regression logistic analysis, the probability of belonging to advanced CTCL stages was associated only with increased cytoplasmic IL-4 (P = 0.007, odds ratio 1.13, 95% confidence interval 1.033-1.229).
Increased T-cell cytoplasmic IL-4 is more frequent in late CTCL stages, correlates with T-cell immunophenotype changes found in advanced disease and is associated with clonality. Increased cytoplasmic IL-4 is frequent both in erythrodermic MF and SS patients, in contrast to other variables found increased only in SS, suggesting that IL-4 may be an early indicator of disease progression. Moreover, our results show that increased cytoplasmic IL-4 is the sole predictor of advanced CTCL disease and confirm the relevance of FC determination of IL-4 in the routine evaluation of CTCL cases.
有证据表明，在 Sézary 综合征（SS）患者的外周血以及组织中，辅助性 T 细胞（Th）2 细胞因子模式占主导地位，且恶性克隆具有 Th2 表型。然而，关于皮肤 T 细胞淋巴瘤（CTCL）不同阶段外周血细胞因子模式的研究结果存在矛盾。
通过流式细胞术（FC）检测蕈样肉芽肿（MF）和 SS（CTCL 最常见的形式）患者外周血 T 细胞中的 Th1/Th2 细胞因子谱[细胞质干扰素（IFN）-γ/白细胞介素（IL）-4]，并将其表达与临床分期、克隆性及 T 细胞免疫表型变化相关联，以评估它们在 CTCL 进展中的相关性。
我们通过 FC 检测了 43 例 CTCL 患者（32 例 I-II 期和 11 例 III-IV 期）血液标本中经刺激后表达细胞质 IFN-γ和 IL-4 的 CD3⁺T 细胞百分比，其中 8 例为红皮病型。接下来，我们比较了不同阶段患者与对照组之间细胞质 IFN-γ和 IL-4 的表达，并将我们的发现与用作克隆性标志物的 T 细胞受体（TCR）-γ基因重排以及 T 细胞免疫表型（CD4⁺、CD8⁺、CD4⁺/CD7⁻、CD4⁺/CD25⁺）和自然杀伤细胞的变化相关联。在 41 份血液和 26 份皮肤标本中进行了 TCR-γ基因的聚合酶链反应扩增。我们还检测了红皮病型 MF 和 SS 患者的细胞因子表达模式。
与早期（I-II 期）CTCL 患者（P = 0.002）或对照组（P < 0.001）相比，晚期（III-IV 期）CTCL患者中 CD3⁺/IL-4⁺T 细胞的频率显著更高。CD3⁺/IL-4⁺百分比与 CD3⁺/CD4⁺T 细胞百分比（r = 0.385，P = 0.05）、CD4⁺/CD7⁻T 细胞百分比（r = 0.335，P < 0.05）和 CD4⁺/CD25⁺T 细胞百分比（r = 0.433，P = 0.01）之间存在显著正相关；CD3⁺/IL-4⁺百分比与 CD3⁺/CD8⁺T 细胞百分比之间存在负相关（r = -0.463，P = 0.005），CD3⁺/IFN-γ⁺百分比与 CD3⁺/CD8⁺T 细胞百分比之间存在正相关（r = 0.368，P = 0.02）。CD3⁺/IL-4⁺、CD3⁺/CD4⁺和 CD4⁺/CD7⁻T 淋巴细胞百分比增加与克隆性的存在相关（分别为 P = 0.025、P < 0.001 和 P = 0.0031）。除细胞质 IL-4 外，所有独立变量在 SS 与红皮病型 MF 患者或对照组之间均显示出统计学上的显著差异，与对照组相比，红皮病型 MF 和 SS 患者的细胞质 IL-4 均较高（分别为 P = 0.003 和 P = 0.008）。在多元回归逻辑分析中，属于晚期 CTCL 阶段的概率仅与细胞质 IL-4 增加相关（P = 0.007，优势比 1.13，95%置信区间 1.
033 - 1.229）。
晚期 CTCL 阶段 T 细胞细胞质 IL-4 增加更为常见，与晚期疾病中发现的 T 细胞免疫表型变化相关且与克隆性有关。红皮病型 MF 和 SS 患者中细胞质 IL-4 增加均很常见，与仅在 SS 中发现增加的其他变量不同，这表明 IL-4 可能是疾病进展的早期指标。此外，我们的结果表明细胞质 IL-4 增加是晚期 CTCL 疾病的唯一预测指标，并证实了 FC 测定 IL-4 在 CTCL 病例常规评估中的相关性。
