Association Analysis of HLA-Class II and Class III Gene Polymorphisms in the Susceptibility to Mediterranean Visceral Leishmaniasis A. Meddeb-Garnaoui, S. Gritli, S. Garbouj, M. Ben Fadhel, R. El Kares, L. Mansour, B. Kaabi, L. Chouchane, A. Ben Salah, and K. Dellagi ABSTRACT: HLA-DRB1, -DQB1, TNF, TNF, HSP70-2 and HSP70-hom genetic polymorphisms were an- alyzed in 156 unrelated patients who developed mediterra- nean visceral leishmaniasis (MVL) due to Leishmania infan- tum, and 154 unrelated healthy controls, who have got asymptomatic infection with this parasite and were selected on the basis of a positive leishmanin skin test (LST). A signifi- cantly reduced frequency of HLA-DR2 was observed among MVL patients (16.1%), compared with controls (26.3%) (rel- ative risk = 0.54; p = 0.04). HLA-DR2/DR13 as well as HLA-DQB1*0201/- genotype frequencies were significantly lower in patients vs controls (relapse rate = 0.17 and 0.46, respectively; p  0.05). However, using Bonferroni correc- tion, none of these associations remained significant. No as- sociation was found, between either the -308 base pair TNF gene polymorphism or the NcoI polymorphism in the first intron of the TNF gene and susceptibility to MVL. Analysis of PstI and NcoI polymorphisms in the coding region of HSP70-2 and HSP70-hom genes, respectively, re- vealed a significantly higher frequency of homozygotes for the HSP70-2/PstI negative allele, among patients (21.8%) vs controls (12.6%) (relapse rate = 1.94; p = 0.04). Again, this result was not significant after using Bonferroni correction. These results do not support association between susceptibil- ity to MVL and the MHC class II and class III loci analyzed in this study. Human Immunology 62, 509 –517 (2001). © American Society for Histocompatibility and Immu- nogenetics, 2001. Published by Elsevier Science Inc. KEYWORDS: MHC genes; Leishmaniasis; tumor necro- sis factor; genetic susceptibility; HLA; DNA typing; can- didate gene ABBREVIATIONS MHC major histocompatibility complex MVL mediterranean visceral leishmaniasis LST leishmanin skin test RR relative risk. TNF tumor necrosis factor HSP70 heat shock protein HLA human leukocyte antigen DTH delayed type hypersensitivity PCR-SSO PCR-sequence specific oligonucleotide TDT transmission disequilibrium test INTRODUCTION Human leishmaniasis are characterized by a wide spec- trum of disease phenotypes ranging from asymptomatic infection to cutaneous, mucocutaneous, or visceral dis- ease. This heterogeneity reflects the complexity of the host-parasite interactions, in which the genetics of the parasite, the genetic background of the host, and the host’s immune response play a major role [1, 2]. Medi- terranean visceral leishmaniasis (MVL) is caused by L. infantum parasites and mainly affects children  5 years old in developing countries [3, 4]. In endemic areas asymptomatic infection is the rule and it is associated From the Laboratory of Immunology (A.M.-G., S.G., S.G., M.B.F., R.E.K., L.M,.B.K., K.D.) and Laboratory of Epidemiology and Ecology of Parasitic Diseases (A.B.S.), Institut Pasteur de Tunis, Tunis-Belve ´de`re, Tunisia; Department of Epidemiology and Biostatistics (B.K.), School of Medicine, Case Western Reserve University, Cleveland, OH, USA; Labora- tory of Immunology (L.C.), Faculty of Medicine, Monastir, Tunisia. Address reprint requests to: Dr. A. Meddeb-Garnaoui, Laboratory of Immunology, Institut Pasteur de Tunis, 13, Place Pasteur-BP74, 1002 Tunis-Belve ´de`re, Tunisia; Tel: +216 (1) 843-755; Fax: +216 (1) 791-833; E-mail: amel.garnaoui@pasteur.rns.tn. Received August 11, 2000; revised January 10, 2001; accepted February 12, 2001. Human Immunology 62, 509 –517 (2001) 0198-8859/01/$–see front matter © American Society for Histocompatibility and Immunogenetics, 2001 Published by Elsevier Science Inc. PII S0198-8859(01)00237-3