Bacherini Daniela, Maggi Laura, Faraldi Francesco, Sodi Andrea, Vannozzi Lorenzo, Mazzoni Alessio, Capone Manuela, Virgili Gianni, Vicini Giulio, Falsini Benedetto, Cosmi Lorenzo, Viggiano Pasquale, Rizzo Stanislao, Annunziato Francesco, Giansanti Fabrizio, Liotta Francesco
Department of Neurosciences, Psychology, Drug Research and Child Health Eye Clinic, University of Florence, AOU Careggi, 50139 Florence, Italy.
Department of Experimental and Clinical Medicine, University of Florence, 50134 Florence, Italy.
Int J Mol Sci. 2024 Dec 7;25(23):13163. doi: 10.3390/ijms252313163.
Recently, evidence has supported a significant role for immune and oxidative-mediated damage underlying the pathogenesis of different types of retinal diseases, including retinitis pigmentosa (RP). Our study aimed to evaluate the presence of immune cells and mediators in patients with RP using flow cytometric analysis of peripheral blood (PB) and aqueous humor (AH) samples. We recruited 12 patients with RP and nine controls undergoing cataract surgery. Flow cytometric analysis of PB and AH samples provided a membrane staining that targeted surface molecules (CD14, CD16, CD19, CD3, CD4, CD8, and CD161) identifying monocytes, natural killer (NK) cells, B cells, T cells, and T subpopulations, respectively. Moreover, lymphocytes were polyclonally stimulated to evaluate cytokine (CK) production at single-cell level. The circulating immune cell distribution was comparable between patients with RP and controls. Conversely, in the AH of controls we could detect no cells, while in the RP AH samples we found infiltrating leukocytes, consisting of T (CD3+), B (CD19+), NK (CD16+CD3-) cells, and monocytes (CD14+). In patients with RP, the frequency of most infiltrating immune cell populations was similar between the AH and PB. However, among T cell subpopulations, the frequency of CD3+CD4+ T cells was significantly lower in the RP AH compared to RP PB, whereas CD3+CD4-CD8- double-negative (DN) T cells were significantly higher in the RP AH compared to RP PB. Cytokine production analysis revealed a trend toward an increased frequency of CD3+CD8-CD161+IFN-ɣ-producing cells and a decreased frequency of CD3+CD8+IL-4-producing cells in the RP AH compared to RP PB. The detection of immune cells, particularly DN T cells, and a Th1-skewed phenotype in RP AH suggests immune-mediated and inflammatory mechanisms in the disease.
最近，有证据支持免疫和氧化介导的损伤在包括色素性视网膜炎（RP）在内的不同类型视网膜疾病发病机制中起重要作用。我们的研究旨在通过对外周血（PB）和房水（AH）样本进行流式细胞术分析，评估RP患者体内免疫细胞和介质的存在情况。我们招募了12例RP患者和9例接受白内障手术的对照者。对PB和AH样本进行流式细胞术分析，提供了一种针对表面分子（CD14、CD16、CD19、CD3、CD4、CD8和CD161）的膜染色，分别识别单核细胞、自然杀伤（NK）细胞、B细胞、T细胞和T亚群。此外，淋巴细胞受到多克隆刺激，以评估单细胞水平上细胞因子（CK）的产生。RP患者和对照者的循环免疫细胞分布具有可比性。相反，在对照者的AH中我们未检测到细胞，而在RP的AH样本中我们发现有浸润的白细胞，包括T（CD3+）、B（CD19+）、NK（CD16+CD3-）细胞和单核细胞（CD14+）。在RP患者中，大多数浸润免疫细胞群体在AH和PB中的频率相似。然而，在T细胞亚群中，与RP的PB相比，RP的AH中CD3+CD4+T细胞的频率显著降低，而CD3+CD4-CD8-双阴性（DN）T细胞在RP的AH中比RP的PB显著更高。细胞因子产生分析显示，与RP的PB相比，RP的AH中CD3+CD8-CD161+IFN-γ产生细胞的频率有增加趋势，而CD3+CD8+IL-4产生细胞的频率有降低趋势。在RP的AH中检测到免疫细胞，特别是DN T细胞，以及Th1偏向的表型，提示该疾病存在免疫介导和炎症机制。
