Department of Zoology, Centre for Advanced Study, Savitribai Phule Pune University, Pune, 411007, India.
INNODx Solutions Pvt Ltd, Biotech Bioincubator (BBB), Regional Centre for Biotechnology, NCR Biotech Science Cluster, Faridabad, Haryana, 1221001, India.
Acta Parasitol. 2022 Jun;67(2):687-696. doi: 10.1007/s11686-021-00511-3. Epub 2022 Jan 12.
The trypanosomatid protozoan parasite Leishmania donovani is the etiological agent of visceral leishmaniasis (VL) or kala-azar. The patients that have undergone treatment may still harbor the parasite and in a small fraction of the patients the disease re-erupts in the form of post kala-azar dermal leishmaniasis (PKDL). PKDL is a pathological condition found to be intermediate between VL and complete cure of VL. The PKDL disease progression is determined by the host immune response to L. donovani. The majority of the proteomic studies on L. donovani till date have been undertaken on parasites either isolated from kala-azar patients or on established laboratory strains of L. donovani. However, no proteomic information is available on the cutaneous localized isolates of L. donovani from PKDL patients.
The promastigote stage of L. donovani isolate from PKDL patient was cultured and harvested. The cell lysates were trypsin digested, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The LC-MS/MS raw data were analyzed on Proteome Discoverer. Further bioinformatics analysis was carried out.
In the present, we have used high-resolution mass spectrometry to map the global proteome of a L. donovani isolate from PKDL patient. This in-depth study resulted in the identification of 5537 unique proteins from PKDL isolate of L. donovani which covered 64% of its proteome.
This study also identified proteins previously shown to be upregulated in PKDL L. donovani. This is the most in-depth proteome of Leishmania donovani parasite till date.
锥虫原生动物寄生虫利什曼原虫是内脏利什曼病（VL）或黑热病的病原体。接受过治疗的患者体内仍可能携带寄生虫，在一小部分患者中，疾病会以皮肤利什曼病（PKDL）的形式重新爆发。PKDL 是一种介于 VL 和 VL 完全治愈之间的病理状态。PKDL 疾病的进展取决于宿主对利什曼原虫的免疫反应。迄今为止，对利什曼原虫的大多数蛋白质组学研究都是在从黑热病患者中分离出的寄生虫或已建立的利什曼原虫实验室株上进行的。然而，对于来自 PKDL 患者的皮肤局限性利什曼原虫分离株，尚未有蛋白质组学信息。
培养并收获来自 PKDL 患者的利什曼原虫前鞭毛体阶段。对细胞裂解物进行胰蛋白酶消化，然后进行液相色谱-串联质谱（LC-MS/MS）分析。在 Proteome Discoverer 上分析 LC-MS/MS 原始数据。进一步进行了生物信息学分析。
目前，我们使用高分辨率质谱技术绘制了来自 PKDL 患者的利什曼原虫分离株的全蛋白质组图谱。这项深入研究确定了来自 PKDL 利什曼原虫分离株的 5537 个独特蛋白质，覆盖了其蛋白质组的 64%。
这项研究还鉴定了以前在 PKDL 利什曼原虫中显示上调的蛋白质。这是迄今为止对利什曼原虫寄生虫最深入的蛋白质组学研究。
