Chinmayee Bar Routaray1, Avishek Kumar2, Shyam Sundar3, Gajanan Sathe4,5, Harsh Pawar6,7, Kalpana Pai8. 1. Department of Zoology, Centre for Advanced Study, Savitribai Phule Pune University, Pune, 411007, India. 2. INNODx Solutions Pvt Ltd, Biotech Bioincubator (BBB), Regional Centre for Biotechnology, NCR Biotech Science Cluster, Faridabad, Haryana, 1221001, India. 3. Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005, India. 4. Institute of Bioinformatics, International Technology Park, Bangalore, 560066, India. gajanan@ibioinformatics.org. 5. Manipal Academy of Higher Education, Manipal, India. gajanan@ibioinformatics.org. 6. Faculty of Biology, Technion-Israel Institute of Technology, 3200003, Haifa, Israel. harshpawar@campus.technion.ac.il. 7. Division of Biomedical & Life Sciences, Faculty of Health & Medicine, Lancaster University, Lancaster, UK. harshpawar@campus.technion.ac.il. 8. Department of Zoology, Centre for Advanced Study, Savitribai Phule Pune University, Pune, 411007, India. drkalpanapai@gmail.com.
Abstract
BACKGROUND: 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. METHODS: 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. RESULTS: 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. OUTCOME: 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.
BACKGROUND: 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. METHODS: 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. RESULTS: 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. OUTCOME: 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.