Deepak Deep - Academia.edu (original) (raw)

Papers by Deepak Deep

* Total cases recruited were 86 of which 73 completed treatment since 12 dropped out and... more * Total cases recruited were 86 of which 73 completed treatment since 12 dropped out and 1 was unresponsive to MIL therapy.Abbreviations: HR, hyper-endemic areas; MR, meso-endemic areas; LR, low-endemic areasProfile of patients treated with miltefosine and relapsed cases.<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004093#t002fn001" target="_blank">*</a></p

Recruitment, treatment and follow-up of PKDL patients.</p

PLOS Neglected Tropical Diseases, 2015

Case definition for recording treatment outcome of PKDL patients.</p

PLOS Neglected Tropical Diseases, 2015

Parasite load was determined by Q-PCR in slit aspirate sample at the time of diagnosis o... more Parasite load was determined by Q-PCR in slit aspirate sample at the time of diagnosis of PKDL and expressed as the number of Leishmania parasite/μl slit aspirate. P value was calculated using Mann-Whitney test. Horizontal bars indicate mean± SEM.</p

PLOS Neglected Tropical Diseases, 2015

Papulo-nodules present on face before treatment (A), and complete regression after 3 mon... more Papulo-nodules present on face before treatment (A), and complete regression after 3 months of MIL treatment (B), relapse at 13 months after completion of MIL treatment with papules on the root of nose (C), and cured after treatment with AmB (D).</p

PLoS neglected tropical diseases, Jan 2, 2017

Miltefosine (MIL) is an oral antileishmanial drug used for treatment of visceral leishmaniasis (V... more Miltefosine (MIL) is an oral antileishmanial drug used for treatment of visceral leishmaniasis (VL) in the Indian subcontinent. Recent reports indicate a significant decline in its efficacy with a high rate of relapse in VL as well as post kala-azar dermal leishmaniasis (PKDL). We investigated the parasitic factors apparently involved in miltefosine unresponsiveness in clinical isolates of Leishmania donovani. L. donovani isolated from patients of VL and PKDL at pretreatment stage (LdPreTx, n = 9), patients that relapsed after MIL treatment (LdRelapse, n = 7) and parasites made experimentally resistant to MIL (LdM30) were included in this study. MIL uptake was estimated using liquid chromatography coupled mass spectrometry. Reactive oxygen species and intracellular thiol content were measured fluorometrically. Q-PCR was used to assess the differential expression of genes associated with MIL resistance. LdRelapse parasites exhibited higher IC50 both at promastigote level (7.92 ± 1.30...

International Journal of Infectious Diseases, 2016

Conclusion: Together, our findings indicate dynamic changes to macrophage and monocytes populatio... more Conclusion: Together, our findings indicate dynamic changes to macrophage and monocytes populations in VL patients over the course of drug treatment, and suggest that the functions of these cells may change at different stages of disease. We found upregulation of some markers for monocyte deactivation.

Clinical & Experimental Immunology, 2016

Summary Post-kala-azar dermal leishmaniasis (PKDL) is a chronic dermal complication that occurs u... more Summary Post-kala-azar dermal leishmaniasis (PKDL) is a chronic dermal complication that occurs usually after recovery from visceral leishmaniasis (VL). The disease manifests into macular, papular and/or nodular clinical types with mono- or polymorphic presentations. Here, we investigated differences in immunological response between these two distinct clinical forms in Indian PKDL patients. Peripheral blood mononuclear cells of PKDL and naive individuals were exposed in vitro to total soluble Leishmania antigen (TSLA). The proliferation index was evaluated using an enzyme-linked immunosorbent assay (ELISA)-based lymphoproliferative assay. Cytokines and granzyme B levels were determined by cytometric bead array. Parasite load in tissue biopsy samples of PKDL was quantified by quantitative polymerase chain reaction (qPCR). The proportion of different lymphoid subsets in peripheral blood and the activated T cell population were estimated using flow cytometry. The study demonstrated he...

PLOS Neglected Tropical Diseases, 2015

Background Recent studies have shown significant decline in the final cure rate after miltefosine... more Background Recent studies have shown significant decline in the final cure rate after miltefosine treatment in visceral leishmaniasis. This study evaluates the efficacy of miltefosine in the treatment of post kala-azar dermal leishmaniasis (PKDL) patients recruited over a period of 5 years with 18 months of follow-up. Methodology In this study 86 confirmed cases of PKDL were treated with two different dosage regimens of miltefosine (Regimen I-50mg twice daily for 90 days and Regimen II-50 mg thrice for 60 days) and the clinical outcome assessed monthly. Cure/relapse was ascertained by clinical and histopathological examination, and measuring parasite burden by quantitative real-time PCR. In vitro susceptibility of parasites towards miltefosine was estimated at both promastigote and amastigote stages. Results Seventy three of eighty six patients completed the treatment and achieved clinical cure. Approximately 4% (3/73) patients relapsed by the end of 12 months follow-up, while a total of 15% (11/73) relapsed by the end of 18 months. Relapse rate was significantly higher in regimen II (31%) compared to regimen I (10.5%)(P<0.005). Parasite load at the pre-treatment stage was significantly higher (P<0.005) in cases that relapsed compared to the cases that remained cured. In vitro susceptibility towards miltefosine of parasites isolated after relapse was significantly lower (>2 fold) in comparison with the pre-treatment isolates (P<0.005). Conclusion Relapse rate in PKDL following miltefosine treatment has increased substantially, indicating the need of introducing alternate drugs/ combination therapy with miltefosine.

Background Miltefosine (MIL) is an oral antileishmanial drug used for treatment of visceral leish... more Background Miltefosine (MIL) is an oral antileishmanial drug used for treatment of visceral leishmaniasis (VL) in the Indian subcontinent. Recent reports indicate a significant decline in its efficacy with a high rate of relapse in VL as well as post kala-azar dermal leishmaniasis (PKDL). We investigated the parasitic factors apparently involved in miltefosine unresponsiveness in clinical isolates of Leishmania donovani. Methodology L. donovani isolated from patients of VL and PKDL at pretreatment stage (LdPreTx, n = 9), patients that relapsed after MIL treatment (LdRelapse, n = 7) and parasites made experimentally resistant to MIL (LdM30) were included in this study. MIL uptake was estimated using liquid chromatography coupled mass spectrometry. Reactive oxygen species and intracellular thiol content were measured fluorometrically. Q-PCR was used to assess the differential expression of genes associated with MIL resistance. Results LdRelapse parasites exhibited higher IC 50 both at promastigote level (7.92 ± 1.30 μM) and at intracellular amastigote level (11.35 ± 6.48 μM) when compared with LdPreTx parasites (3.27 ± 1.52 μM) and (3.85 ± 3.11 μM), respectively. The percent infectivity (72 hrs post infection) of LdRelapse parasites was significantly higher (80.71 ± 5.67%, P<0.001) in comparison to LdPreTx (60.44 ± 2.80%). MIL accumulation was significantly lower in LdRelapse parasites (1.7 fold, P<0.001) and in LdM30 parasites (2.4 fold, P<0.001) when compared

PLoS Neglected Tropical Diseases, 2012

Background: With widespread resistance to antimonials in Visceral Leishmaniasis (VL) in the India... more Background: With widespread resistance to antimonials in Visceral Leishmaniasis (VL) in the Indian subcontinent, Miltefosine (MIL) has been introduced as the first line therapy. Surveillance of MIL susceptibility in natural populations of Leishmania donovani is vital to preserve it and support the VL elimination program. Methodology and Principal Findings: We measured in vitro susceptibility towards MIL and paromomycin (PMM) in L. donovani isolated from VL and PKDL, pre-and post-treatment cases, using an amastigote-macrophage model. MIL susceptibility of post-treatment isolates from cured VL cases (n = 13, mean IC 50 6SD = 2.4361.44 mM), was comparable (p.0.05) whereas that from relapses (n = 3, mean IC 50 = 4.7261.99 mM) was significantly higher (p = 0.04) to that of the pretreatment group (n = 6, mean IC 50 = 1.8660.75 mM). In PKDL, post-treatment isolates (n = 3, mean IC 50 = 16.1362.64 mM) exhibited significantly lower susceptibility (p = 0.03) than pre-treatment isolates (n = 5, mean IC 50 = 8.6360.94 mM). Overall, PKDL isolates (n = 8, mean IC 50 = 11.4564.19 mM) exhibited significantly higher tolerance (p,0.0001) to MIL than VL isolates (n = 22, mean IC 50 = 2.5861.58 mM). Point mutations in the miltefosine transporter (LdMT) and its beta subunit (LdRos3) genes previously reported in parasites with experimentally induced MIL resistance were not present in the clinical isolates. Further, the mRNA expression profile of these genes was comparable in the pre-and post-treatment isolates. Parasite isolates from VL and PKDL cases were uniformly susceptible to PMM with respective mean IC 50 = 7.0562.24 mM and 6.1861.51 mM. Conclusion: The in vitro susceptibility of VL isolates remained unchanged at the end of MIL treatment; however, isolates from relapsed VL and PKDL cases had lower susceptibility than the pre-treatment isolates. PKDL isolates were more tolerant towards MIL in comparison with VL isolates. All parasite isolates were uniformly susceptible to PMM. Mutations in the LdMT and LdRos3 genes as well as changes in the expression of these genes previously correlated with experimental resistance to MIL could not be verified for the field isolates.

Pathogens, 2021

Visceral leishmaniasis (VL), mainly caused by the Leishmania donovani parasitic infection, consti... more Visceral leishmaniasis (VL), mainly caused by the Leishmania donovani parasitic infection, constitutes a potentially fatal disease, for which treatment is primarily dependent on chemotherapy. The emergence of a resistant parasite towards current antileishmanial agents and increasing reports of relapses are the major concerns. Detailed research on the molecular interaction at the host-parasite interface may provide the identification of the parasite and the host-related factors operating during disease development. Genomic and proteomic studies highlighted several essential secretory and cytosolic proteins that play vital roles during Leishmania pathogenesis. The aim of this study was to identify membrane proteins from the Leishmania donovani parasite and the host macrophage that interact with each other using 2-DE/MALDI-TOF/MS. We identified membrane proteins including activated protein C kinase, peroxidoxin, small myristoylated protein 1 (SMP-1), and cytochrome C oxidase from the p...

1WHO code: country and year of isolation and the respective strain code, the number foll... more 1WHO code: country and year of isolation and the respective strain code, the number following the isolate ID indicates the number of months elapsed after start of MIL treatment.2Research code: Parasites cultured from VL patients were labeled V- and from PKDL patients P-, respectively. The number following the isolate ID indicates the number of months elapsed after start of MIL treatment (e.g. V902/1 means one month passed from first MIL treatment). Parasites isolated from patients' prior start of MIL treatment were labeled as XXX/0 and one month following first treatment was labeled XXX/1. These patients cleared from VL symptoms after respective duration of MIL treatment and were interpreted as clinical cure, although residual parasites could be cultured from splenic aspirates (marked XXX/1). In the period of 1 year follow up, cases of relapse were observed in three VL patients that had shown an initial clinical cure, the isolates obtained were design...

In vitro miltefosine susceptibility (IC50), MI... more In vitro miltefosine susceptibility (IC50), MIL accumulation and thiol levels of L. donovani isolates from pretreatment group (LdPreTx), isolates obtained from visceral leishmaniasis (VL) and post kala-azar dermal leishmaniasis (PKDL) patients that relapsed after MIL treatment (LdRelapse) and experimental MIL resistant parasites (LdM30).</p

(A) MIL susceptibility at promastigotes stage of LdPreTx, LdRelapse and LdM30 with each ... more (A) MIL susceptibility at promastigotes stage of LdPreTx, LdRelapse and LdM30 with each individual value represented as mean IC50±SD from three separate assays. (B) MIL susceptibility at intracellular amastigote stage LdPreTx, LdRelapse and LdM30 with each individual value represented as mean IC50 ± SD from three separate assays (C) Mice peritoneal derived macrophages infected with LdPreTx or LdRelapse parasites at a 1:10 (cell/parasite) ratio. The percent infectivity was determined at 24h, 48h, and 72h post infection by counting number of infected cells out of 100 macrophages at 1000X magnification after staining with Diff-Quik. Data represents mean ± SD of three independent experiments each in duplicate. (D) Percent metacyclogenesis of promastigote population estimated based on negative selection of peanut agglutinin (%PNA- promastigote). Values represent mean ± SD of two independent experiments (E) MIL uptake, estimated using LC-MS in 1x108 promastigotes of LdPreTx, LdRelpase and LdM30. Data represents mean ± SD of two independent experiments, each in duplicate. Asterisks indicate significance (**P<0.01; and ***P<0.001).</p

Fold change represents expression of target genes normalized to internal control (GAPDH ... more Fold change represents expression of target genes normalized to internal control (GAPDH and α-Tubulin) genes and relative to LdAG83. Data represents mean ± SD of two separate assays, each performed in triplicate. Asterisks indicate significance (*P<0.05; **P<0.01; and *** P<0.001).</p

(A) Dose dependent accumulation of ROS in LdPreTx, LdRelapse and LdM30 at promastigote s... more (A) Dose dependent accumulation of ROS in LdPreTx, LdRelapse and LdM30 at promastigote stage was assayed fluorometrically at 495 nm excitation and 535 nm emission wavelength using cell permeable probe H2DCF-DA (40nM). Data represents mean ± SD of three independent experiments, each performed in triplicate. Asterisks indicate significance (**P<0.01; and ***P<0.001). (B) Accumulation of ROS in macrophages infected with LdPreTx, LdRelapse, LdM30 parasites before and after MIL exposure (20μM), assayed fluorometrically at 495 nm excitation and 535 nm emission wavelength using cell permeable probe H2DCF-DA (30μM). Data represents mean ± SD of three independent experiments, each in triplicate. Asterisks indicate significance (*P<0.05; **P<0.01; and ***P<0.001). (C) Intracellular thiol content in LdPreTx, LdRelapse and LdM30 promastigotes, measured fluorometrically at 390 nm excitation and 520 nm emission wavelength. Data represents mean ± SD of two independent experiments each performed in triplicate. Asterisks indicate significance (**P<0.01; and ***P<0.001).</p

Susceptibility of VL and PKDL isolates at intracellular amastigote stage was determined ... more Susceptibility of VL and PKDL isolates at intracellular amastigote stage was determined by infection in murine macrophage cell line J774A.1. Each individual value represents mean IC50±SD of the results from two separate assays.</p

PLOS Neglected Tropical Diseases, 2015

MIL susceptibility at (A) promastigote stage (B) amastigote stage. Each individual value... more MIL susceptibility at (A) promastigote stage (B) amastigote stage. Each individual value represents mean IC50± SD of the results from two separate assays. P value was calculated using Mann-Whitney test. Horizontal bars indicate mean ±SEM.</p

Recruitment, treatment and follow-up of PKDL patients.</p

PLOS Neglected Tropical Diseases, 2015

Case definition for recording treatment outcome of PKDL patients.</p

PLOS Neglected Tropical Diseases, 2015

PLoS neglected tropical diseases, Jan 2, 2017

International Journal of Infectious Diseases, 2016

Clinical & Experimental Immunology, 2016

PLOS Neglected Tropical Diseases, 2015

PLoS Neglected Tropical Diseases, 2012

Pathogens, 2021

PLOS Neglected Tropical Diseases, 2015