Monitoring of Intracellular Nitric Oxide in Leishmaniasis: Its Applicability in Patients with Visceral Leishmaniasis
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
View Archive InfoField | Value | |
Title |
Monitoring of Intracellular Nitric Oxide in
Leishmaniasis: Its Applicability in Patients
with Visceral Leishmaniasis
|
|
Creator |
Sarkar, Avijit
Saha , Piu Mandal, Goutam Mukhopadhyay, Debanjan Roy , Susmita Singh, Sunny Kumar Das, Sukhen Goswami, Rama Prosad Saha, Bibhuti Kumar, Deepak Das, Padma Chatterjee, Mitali |
|
Subject |
Cell Biology & Physiology
|
|
Description |
Nitric oxide (NO) has been demonstrated to be a principal effector molecule responsible for mediating intracellular killing of Leishmania parasites, the causative organism
of leishmaniasis. As measurement of intracellular NO remains a challenge to biologists, we have developed a flow cytometric approach to perform real time biological detection of NO within Leishmania parasites and parasitized macrophages
using a membrane permeable derivative of diaminofluorescein [4,5- diaminofluorescein
diacetate (DAF-2DA)]. Initially, assay optimization was performed in Leishmania
donovani promastigotes, assay specificity being confirmed using both a NO donor
[S-nitroso-N-acetyl-penicillamine (SNAP)] and a NO scavenger [2-(4-carboxyphenyl)-
4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, C-PTIO]. Using 40 lM DAF- 2DA, basal levels of intracellular NO were measured which varied in different Leishmania species; addition of conventional anti-leishmanial drugs, antimony and miltefosine translated into a dramatic increase in DAF-2T fluorescence. Furthermore, the assay also measured levels of NO in macrophages, but needed a 20 fold lower concentration of DAF-2DA, being 2 lM. Following parasitization, levels of NO decreased
which was normalized following treatment with anti-leishmanial drugs. Similarlymonocytes of patients with visceral leishmaniasis at disease presentation showeddecreased levels of NO which too reverted on completion of treatment. Taken together,this study opens new perspectives of research regarding monocyte function
and provides a real time approach for monitoring the effect of anti-leishmanial compounds.
|
|
Date |
2011
|
|
Type |
Article
PeerReviewed |
|
Format |
application/pdf
|
|
Identifier |
http://www.eprints.iicb.res.in/55/1/CYTOMETRY%2C_PART%2D_A%2C__79A(1)_35%2D45%2C2011[106].pdf
Sarkar, Avijit and Saha , Piu and Mandal, Goutam and Mukhopadhyay, Debanjan and Roy , Susmita and Singh, Sunny Kumar and Das, Sukhen and Goswami, Rama Prosad and Saha, Bibhuti and Kumar, Deepak and Das, Padma and Chatterjee, Mitali (2011) Monitoring of Intracellular Nitric Oxide in Leishmaniasis: Its Applicability in Patients with Visceral Leishmaniasis. Cytometry Part A, 79A. pp. 35-45. |
|
Relation |
http://dx.doi.org/10.1002/cyto.a.21001
http://www.eprints.iicb.res.in/55/ |
|