Laboratory of Transmission, Control and Immunobiology of Infections, Pasteur Institute of Tunis, Tunis, Tunisia.
Université de Tunis El Manar, Tunis, Tunisia.
Immun Inflamm Dis. 2019 Sep;7(3):95-104. doi: 10.1002/iid3.240. Epub 2019 Apr 17.
Zoonotic cutaneous leishmaniasis (ZCL), due to infection by Leishmania (L). major, is characterized by polymorphic clinical manifestations which could be attributed to the host's immune response. In this study we investigated the involvement of cytotoxic cells on the outcome of the disease.
Expression of granzyme B (GrB), granulysine (Grly), and interferon (IFN)-γ was evaluated within ZCL lesion specimens using the technique of real-time quantitative polymerase chain reaction (RT-qPCR). Immunohistochemical staining was performed using anti-CD3, CD4, CD8, CD56, GrB, and IFN-γ antibodies to identify the phenotype of GrB and IFN-γ-producing cells.
GrB and Grly mRNA was detected within 75% and 80% of ZCL lesions, respectively. Statistical analysis demonstrated a significant correlation between levels of GrB and Grly. Interestingly, expression of these molecules correlates negatively with the lesion's age. The highest levels were measured in early lesions (E-ZCL) (lesion age ≤1 month) comparing to late lesions (L-ZCL) (lesion age >1 month). Otherwise, IFN-γ mRNA was detected only within 56% and a positive correlation was found between levels of this cytokine and those of GrB. Immunohistochemical analysis showed that GrB is produced essentially by CD8 T cells whereas IFN-γ is produced by both CD4 and CD8 T cells.
Together our results demonstrate the presence of cytotoxic cells producing GrB and Grly within leishmaniasis cutaneous lesions.
由于感染了利什曼原虫（L）。主要，动物源性皮肤利什曼病（ZCL）的特征是多态性临床表现，这可能归因于宿主的免疫反应。在这项研究中，我们研究了细胞毒性细胞在疾病结果中的作用。
使用实时定量聚合酶链反应（RT-qPCR）技术评估 ZCL 病变标本中颗粒酶 B（GrB）、颗粒溶素（Grly）和干扰素（IFN）-γ的表达。使用抗 CD3、CD4、CD8、CD56、GrB 和 IFN-γ 抗体进行免疫组织化学染色，以鉴定 GrB 和 IFN-γ 产生细胞的表型。
GrB 和 Grly mRNA 在分别在 75%和 80%的 ZCL 病变中检测到。统计分析表明 GrB 和 Grly 之间存在显著相关性。有趣的是，这些分子的表达与病变的年龄呈负相关。在早期病变（E-ZCL）（病变年龄≤1 个月）中测量到的水平最高，与晚期病变（L-ZCL）（病变年龄>1 个月）相比。另一方面，IFN-γ mRNA 仅在 56%的病变中检测到，并且发现这种细胞因子的水平与 GrB 的水平呈正相关。免疫组织化学分析表明，GrB 主要由 CD8 T 细胞产生，而 IFN-γ 由 CD4 和 CD8 T 细胞产生。
总之，我们的结果表明在皮肤利什曼病病变中存在产生 GrB 和 Grly 的细胞毒性细胞。
