Analysis of genetic variants in the IL4 promoter and VNTR loci in Indian patients with Visceral Leishmaniasis Anshuman Mishra a,1 , Aditya Nath Jha a,g,1 , Hoang van Tong b , Vipin Kumar Singh a , Carlos E.M. Gomes c , Lalji Singh a,d,e , Thirumalaisamy P. Velavan b,f,⇑,2 , Kumarasamy Thangaraj a,⇑,2 a CSIR – Center for Cellular and Molecular Biology, Hyderabad, India b Institute of Tropical Medicine, University of Tübingen, Tübingen, Germany c Universidade Federal do Rio Grande do Norte, Natal, Brazil d Banaras Hindu University, Varanasi, India e Genome Foundation, Hyderabad, India f Fondation Congolaise pour la Recherche Medicale, Brazzaville, People’s Republic of Congo g Sickle Cell Institute Chattisgarh (SCIC), Raipur, India article info Article history: Received 12 April 2014 Accepted 9 October 2014 Available online 19 October 2014 Keywords: Visceral Leishmaniasis Leishmania donovani Interleukin 4 VNTR India abstract Visceral Leishmaniasis (VL) is the most severest form of Leishmaniasis and resistance to infection is mediated by cellular immune responses. Interleukin 4 (IL-4) orchestrates of Th2 and Th1 immune responses during infections. In this study, we aimed to investigate possible association between three functional IL-4 polymorphisms À590C/T (rs2243250), À34C/T (rs2070874) and 70bp VNTR (rs79071878 in intron3) with VL in an Indian cohort comprising of 197 VL patients and 193 healthy con- trols. The three investigated IL-4 polymorphisms were in strong linkage disequilibrium. The investigated IL-4 alleles, genotypes and the reconstructed haplotypes were not significantly distributed between the VL patients and healthy controls. Our study signifies no possible association of functional IL-4 polymor- phisms with Indian VL and postulate other vital genes involved in the IL-4 pathway may provide genetic clues to elucidate of IL-4 regulation and immune-pathogenesis during VL. Ó 2014 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved. 1. Introduction Leishmaniasis is caused by the protozoan parasite of the genus Leishmania, with an estimated 1.3 million new cases recorded annually [1]. Four distinct clinical forms of Leishmaniasis were reported namely, the cutaneous Leishmaniasis (CL), visceral Leish- maniasis (VL), mucocutaneous Leishmaniasis (MCL) and the post kala-azar dermal Leishmaniasis (PKDL) [2]. Among the clinical presentations, the VL contributes to a higher incidence of new cases with 50,000 estimated deaths every year [3]. VL is a major health burden in the Indian sub continent with150 million people living at risk and countries such as India, Nepal and Bangladesh contributes to the 67% of the global disease burden [4–6]. VL caused by Leishmania donovani is endemic in the Indian state of Bihar and accounts for more than 90% of all reported VL cases within India [4] and VL remains fatal when untreated [7]. A multi- tude of host genetic factors were shown to regulate the outcome of VL [8–10]. The gene expression is controlled by an orderly transcriptional process and are regulated by cis-acting DNA elements in the pro- moter region [11]. Therefore, any nucleotide alteration in the pro- moter region is likely to alter the gene expression and function, thus reflecting the level of susceptibility to an infection [11–15]. Additionally, the pathogen driven selection may also potentially alter the primed sequence and can direct to substantial changes in gene expression. Hence, investigation of genetic variations within the promoter regions is of interest. One such established gene locus is the human interleukin 4 (IL-4) promoter that were demonstrated to modulate disease susceptibility [16]. The human IL-4 encoded by the IL-4 gene, is an anti-inflammatory Th2 cyto- kine positioned at chromosome 5q31-33 and is produced by CD4 + Th2 cells, basophils and mast cells. The human IL-4 is http://dx.doi.org/10.1016/j.humimm.2014.10.007 0198-8859/Ó 2014 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved. ⇑ Corresponding authors at: Institute of Tropical Medicine, Wilhelmstraße 27, 72074 Tübingen, Germany. Fax: +49 7071 294684 (T.P. Velavan). Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500 007, India. Fax: +91 40 27160591 (K. Thangaraj). E-mail addresses: velavan@medizin.uni-tuebingen.de (T.P. Velavan), thangs@ccmb.res.in (K. Thangaraj). 1 Authors contributed equally. 2 Shared last authorship. Human Immunology 75 (2014) 1177–1181 Contents lists available at ScienceDirect www.ashi-hla.org journal homepage: www.elsevier.com/locate/humimm