Changes in RNA expression pattern in neutrophils of patients with Indian Visceral leishmaniasis — ASN Events
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  3. Changes in RNA expression pattern in neutrophils of patients with Indian Visceral leishmaniasis

Changes in RNA expression pattern in neutrophils of patients with Indian Visceral leishmaniasis (#330)

SMRITI SHARMA 1 , RICHARD DAVIS , SUSANNE NYLEN , MARY WILSON , SHYAM SUNDAR 1
  1. Department of Medicine, INSTITUTE OF MEDICAL SCIENCES, BANARAS HINDU UNIVERSITY, VARANASI, Uttar Pradesh, India

Aim- Visceral leishmaniasis (VL), is a disease caused by Leishmania donovani in India. Neutrophils act as first line of defense against invading microorganisms and their interaction with other immune cells and environmental signals during disease can influence  overall immune response. We observed HLA DR expressing low density "neutrophil like cells" which stain CD66b in PBMC and whole blood samples in active VL subjects by flowcytometry previously. In continuation to our previous study we also tried to look at  changes in mRNA expression in active VL pre and post treatment (paired) groups and endemic healthy controls (EHC) to understand the contribution of these cells during active disease.

Methods –

Patient Selection:All patients presented with VL symptoms at KAMRC, Muzaffarpur, India were confirmed to be VL positive by detection of amastigotes in splenic aspirates and/or by positive rk39 test and EHCs  were enrolled from local area.

Gene Expression: Human neutrophils were isolated using Militenyi CD66abce microbead kit to get highly pure neutrophils (> 97%). cDNA was prepared using High capacity cDNA archive kit. Real time PCR was performed using ABI-Prism 7500 using specific Sybr Green primers for mRNA of interest.

Results- Differences were observed in expression of cytokine and chemokine mRNAs in the same subjects (paired) in pre and post treatment(n=8). Some, but not all differences mirrored profiles in EHCs (n=5).

 IFN γ,IL10, were significantly up regulated during Active VL  while  cytokine IL1β and βdefensins  and chemokines CXCL8,CCR4,CCR5,CXCR1,CXCL9  were found to be down regulated. We looked at some important markers like MPO, PD1, Arginase 1, HLA DR which were found to be upregulated during active VL. These markers were studied to address functional competency of neutrophils during active VL at mRNA levels.

 Conclusions- Neutrophils from subjects with active disease differ from the healthy subjects and from the same subjects after drug induced cure showing neutrophils during active disease are capable of extensive changes in gene expression and  can influence development of overall immune response. 

  1. Richard E davis, University of IOWA and THE VA MEDICAL CENTER, Iowa City, IA, United States.
  2. Mary Wilson, University of IOWA and THE VA MEDICAL CENTER, Iowa City, IA, United States.
  3. Susanne Nylen, Department of Microbiology tumor and cell biology, Karolinska Institutet, Sweden