Combination of paromomycin and miltefosine promotes TLR4-dependent induction of antileishmanial immune response in vitro

被引:22
作者
Das, Sushmita
Rani, Mukta
Pandey, Krishna
Sahoo, Ganesh Chandra
Rabidas, Vidya Nand
Singh, Dharmendra
Das, Pradeep [1 ]
机构
[1] ICMR, Rajendra Mem Res Inst Med Sci, Dept Mol Parasitol, Patna, Bihar, India
关键词
TLR4drug interaction; combination therapy; TNF-; nitric oxide; VISCERAL LEISHMANIASIS;
D O I
10.1093/jac/dks220
中图分类号
R51 [传染病];
学科分类号
100401 ;
摘要
To evaluate the in vitro activity of antileishmanial drugs, paromomycin and miltefosine, to generate Th-1-biased immunomodulation in hosts against intracellular Leishmania donovani. In silico proteinligand interaction and in vitro drugcell interaction assays were performed. Interaction assays of TLR4-deficient HEK293 cells and HEK293 cells engineered to express either TLR4 or TLR2 with different concentrations of miltefosine and/or paromomycin sulphate were performed for 48 h. Differentially transfected human peripheral blood monocyte-derived macrophages (PBMFs) were treated with the drugs, and nuclear factor (NF)-B promoter activity was measured using a B-luciferase reporter construct. PBMFs were infected with L. donovani. Cultures were incubated with miltefosine or paromomycin sulphate over different concentrations, as mono-treatment or combined. The in vitro antileishmanial effect of the drugs on macrophage-bound L. donovani amastigotes was measured in terms of parasite killing and production of tumour necrosis factor- (TNF-) and nitric oxide. Computational studies reveal that paromomycin and miltefosine interact with TLR4. Both drugs, as monotherapy or in combination, induce release of TNF- and nitric oxide in a TLR4-dependent manner. Interestingly, the TLR4-dependent action of the drugs leads to NF-B promoter activation through MyD88. In vitro, both the drugs kill macrophage-bound L. donovani by inducing release of TNF- and nitric oxide in a TLR4-dependent manner. The in vitro activity of paromomycin and miltefosine against host cells is TLR4 dependent. This has implications for: (i) evaluation of in vitro activity of combinational antileishmanial therapy; (ii) the evaluation of drug susceptibility of clinical isolates; and (iii) the standardization of in vitro antileishmanial assays for determining toxicity in hosts.
引用
收藏
页码:2373 / 2378
页数:6
相关论文
共 8 条
[1]   Toll-like receptors in the induction of the innate immune response [J].
Aderem, A ;
Ulevitch, RJ .
NATURE, 2000, 406 (6797) :782-787
[2]   Visceral leishmaniasis: What are the needs for diagnosis, treatment and control? [J].
Chappuis, Francois ;
Sundar, Shyam ;
Hailu, Asrat ;
Ghalib, Hashim ;
Rijal, Suman ;
Peeling, Rosanna W. ;
Alvar, Jorge ;
Boelaert, Marleen .
NATURE REVIEWS MICROBIOLOGY, 2007, 5 (11) :873-882
[3]   RETRACTED: TGF-β1 re-programs TLR4 signaling in L. donovani infection: enhancement of SHP-1 and ubiquitin-editing enzyme A20 (Retracted article. See vol. 101, pg. 91, 2023) [J].
Das, Sushmita ;
Pandey, Krishna ;
Kumar, Ashish ;
Sardar, Abul H. ;
Purkait, Bidyut ;
Kumar, Manish ;
Kumar, Sudeep ;
Ravidas, Vidya N. ;
Roy, Syamal ;
Singh, Dharmendra ;
Das, Pradeep .
IMMUNOLOGY AND CELL BIOLOGY, 2012, 90 (06) :640-654
[4]   Visceral leishmaniasis: elimination with existing interventions [J].
Matlashewski, Greg ;
Arana, Byron ;
Kroeger, Axel ;
Battacharya, Sujit ;
Sundar, Shyam ;
Das, Pradeep ;
Sinha, Prabhat Kumar ;
Rijal, Suman ;
Mondal, Dinesh ;
Zilberstein, Dan ;
Alvar, Jorge .
LANCET INFECTIOUS DISEASES, 2011, 11 (04) :322-325
[5]   The antifungal drug amphotericin B promotes inflammatory cytokine release by a Toll-like receptor- and CD14-dependent mechanism [J].
Sau, K ;
Mambula, SS ;
Latz, E ;
Henneke, P ;
Golenbock, DT ;
Levitz, SM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (39) :37561-37568
[6]   In vitro activity of anti-leishmanial drugs against Leishmania donovani is host cell dependent [J].
Seifert, Karin ;
Escobar, Patricia ;
Croft, Simon L. .
JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY, 2010, 65 (03) :508-511
[7]   Oral miltefosine for Indian visceral leishmaniasis [J].
Sundar, S ;
Jha, TK ;
Thakur, CP ;
Engel, J ;
Sindermann, H ;
Fischer, C ;
Junge, K ;
Bryceson, A ;
Berman, J .
NEW ENGLAND JOURNAL OF MEDICINE, 2002, 347 (22) :1739-1746
[8]   Comparison of short-course multidrug treatment with standard therapy for visceral leishmaniasis in India: an open-label, non-inferiority, randomised controlled trial [J].
Sundar, Shyam ;
Sinha, Prabhat Kumar ;
Rai, Madhukar ;
Verma, Deepak Kumar ;
Nawin, Kumar ;
Alam, Shanawwaj ;
Chakravarty, Jaya ;
Vaillant, Michel ;
Verma, Neena ;
Pandey, Krishna ;
Kumari, Poonam ;
Lal, Chandra Shekhar ;
Arora, Rakesh ;
Sharma, Bhawna ;
Ellis, Sally ;
Strub-Wourgaft, Nathalie ;
Balasegaram, Manica ;
Olliaro, Piero ;
Das, Pradeep ;
Modabber, Farrokh .
LANCET, 2011, 377 (9764) :477-486