Exposure to Holoendemic Malaria Results in Suppression of Epstein-Barr Virus-Specific T Cell Immunosurveillance in Kenyan Children

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Exposure to Holoendemic Malaria Results in Suppression of Epstein-Barr Virus-Specific T Cell Immunosurveillance in Kenyan Children University of Massachusetts Medical School eScholarship@UMMS Population and Quantitative Health Sciences Publications Population and Quantitative Health Sciences 2007-02-15 Exposure to holoendemic malaria results in suppression of Epstein-Barr virus-specific T cell immunosurveillance in Kenyan children Ann M. Moormann University of Massachusetts Medical School Et al. Let us know how access to this document benefits ou.y Follow this and additional works at: https://escholarship.umassmed.edu/qhs_pp Part of the Biostatistics Commons, Epidemiology Commons, Health Services Research Commons, Immunology and Infectious Disease Commons, and the Pediatrics Commons Repository Citation Moormann AM, Chelimo K, Sumba PO, Tisch DJ, Rochford RA, Kazura JW. (2007). Exposure to holoendemic malaria results in suppression of Epstein-Barr virus-specific T cell immunosurveillance in Kenyan children. Population and Quantitative Health Sciences Publications. https://doi.org/10.1086/ 511984. Retrieved from https://escholarship.umassmed.edu/qhs_pp/397 This material is brought to you by eScholarship@UMMS. It has been accepted for inclusion in Population and Quantitative Health Sciences Publications by an authorized administrator of eScholarship@UMMS. For more information, please contact [email protected]. MAJOR ARTICLE Exposure to Holoendemic Malaria Results in Suppression of Epstein-Barr Virus–Specific T Cell Immunosurveillance in Kenyan Children Ann M. Moormann,1 Kiprotich Chelimo,4 Peter O. Sumba,4 Daniel J. Tisch,2 Rosemary Rochford,3 and James W. Kazura1 1Center for Global Health and Diseases and 2Department of Epidemiology and Biostatistics, Case Western Reserve University, Cleveland, Ohio; 3Department of Microbiology and Immunology, State University of New York Upstate Medical University, Syracuse; 4Kenya Medical Research Institute, Center for Vector Biology and Control Research, Kisumu, Kenya Background. Malaria and Epstein-Barr virus (EBV) infection are cofactors in the pathogenesis of endemic Burkitt lymphoma (eBL). The mechanisms by which these pathogens predispose to eBL are not known. Methods. Healthy Kenyan children with divergent malaria exposure were measured for responses to EBV latent and lytic antigens by interferon (IFN)–g enzyme-linked immunospot (ELISPOT) assay and interleukin (IL)–10 ELISA. Phytohemagglutinin (PHA), purified protein derivative (PPD), and T cell epitope peptides derived from merozoite surface protein (MSP)–1, a malaria blood-stage antigen, were also evaluated. Results. Children 5–9 years old living in an area holoendemic for malaria had significantly fewer EBV-specific IFN-g responses than did children of the same age living in an area with unstable malaria transmission. This effect was not observed for children !5 years old or those 19 years old. In contrast, IFN-g responses to PHA, PPD, and Plasmodium falciparum MSP-1 peptides did not significantly differ by age. IL-10 responses to EBV lytic antigens, PPD, and PHA correlated inversely with malaria exposure regardless of age. Conclusions. Children living in malaria-holoendemic areas have diminished EBV-specific T cell immuno- surveillance between the ages of 5 and 9 years, which coincides with the peak age incidence of eBL. Burkitt lymphoma was first described in African chil- magenesis. In fact, there is a strong correlation between dren in 1958 [1]. The highest incidence of endemic residence in areas of intense, perennial malaria trans- Burkitt lymphoma (eBL) is in equatorial Africa and mission (i.e., holoendemic malaria) and the incidence Papua New Guinea (5–15 cases/100,000 children) [2, of eBL [3, 5]. 3]. Within Africa, there is an uneven geographic dis- eBL, the first human cancer discovered to have a viral tribution of eBL, which led Dalldorf [4] in 1962 to etiology, is associated with Epstein-Barr virus (EBV) suggest that malaria may be a risk factor for lympho- infection [6]. EBV is restricted to humans and infects the vast majority of the world’s population [7]. In coun- tries such as Kenya, EBV infection generally occurs by Received 1 August 2006; accepted 9 November 2006; electronically published 2 years of age [8–11] and persists for life as a latent 6 February 2007. infection in memory B cells [12]. The lytic phase of Potential conflicts of interest: none reported. Presented in part: American Society of Tropical Medicine and Hygiene 52nd viral replication is thought to occur after plasma cell Annual Meeting, Philadelphia, PA, 3–7 December 2003 (abstract 826); 11th In- differentiation [13–15]. The virus is periodically shed ternational Symposium on EBV and Associated Diseases, University of Regens- burg, Germany, 20–25 September 2004 (abstract 07.05). in the saliva, suggesting that there is ongoing lytic-cycle Financial support: National Institutes of Health (grant K08 AI51565 to A.M., reactivation [16]. R01 CA102667 to R.R., and UO1 AI43906 to J.K.). Immunosurveillance and control of EBV is domi- This article was approved by the director of the Kenya Medical Research + Institute. nated by CD8 human leukocyte antigen (HLA) class Reprints or correspondence: Dr. Ann M. Moormann, Center for Global Health I–restricted cytotoxic T lymphocyte (CTL) interferon and Diseases, Case Western Reserve University, WRB 4-130, 2103 Cornell Rd., Cleveland, OH 44106-7286 ([email protected]). (IFN)–g responses to both latent and lytic viral epitopes The Journal of Infectious Diseases 2007;195:799–808 [17, 18]. An EBV-specific CD8+ T cell subset that has ᮊ 2007 by the Infectious Diseases Society of America. All rights reserved. reduced cytotoxicity and that secretes interleukin (IL)– 0022-1899/2007/19506-0007$15.00 DOI: 10.1086/511984 10 has also been described [19]. Two possible but not EBV Immunity in Malaria-Exposed Children • JID 2007:195 (15 March) • 799 mutually exclusive models have been proposed to explain how PARTICIPANTS, MATERIALS, AND METHODS holoendemic malaria could affect EBV latency and immunity in children and thereby increase the risk of eBL: suppression Participants. Approval for this study was obtained from the of EBV-specific T cell immunity and/or expansion of the la- Kenya Medical Research Institute (KEMRI) National Ethical tently infected B cell pool (reviewed in [20, 21]). The few studies Review Committee and the Institutional Review Board for Hu- that have investigated the role played by malaria in T cell con- man Studies at the University Hospitals of Cleveland, Case trol of EBV-infected B cells in clinical samples have used an in Western Reserve University. Written, informed consent was ob- vitro regression assay that measures the ability of T cells to tained from the guardians or parents of the study participants. control the outgrowth of EBV-transformed B lymphocytes [22– Study participant recruitment and sample collection were 24]; regression of colonies is thought to be mediated by T cells conducted in July–August 2002 in 2 epidemiologically distinct areas of Kenya (Kisumu District and Nandi District), as de- that express CD8 and produce IFN-g [25, 26]. Using this assay, scribed elsewhere [11]. The characteristics of the study popu- Moss et al. [23] found that adults living in malaria-holoendemic lation are summarized in table 1. Children from the holoen- regions of Papua New Guinea had impaired EBV-specific T cell demic-malaria transmission area are referred to as “Kisumu responses. Later observations from The Gambia reported that children” (n p 104 ), and children from the sporadic, unstable children experiencing acute clinical malaria had impaired EBV- malaria transmission area are referred to as “Nandi children” specific T cell immunity [22, 24]. However, conclusions from (n p 127 ). Children (1–14 years old) were enrolled if they had this earlier work were based on a relatively small number of overall good health and were excluded from analysis if they children or on adults with mature T cell immunity, whose were EBV seronegative [11]. Of note, 77% of the Kisumu chil- responses might differ from those of children. In addition, dren had asymptomatic Plasmodium falciparum infection de- regression assays do not identify the effector T cells or cytokine tected by blood smear, which is typical for resident children mediators involved in the control of EBV-infected B cells, and from this area. In contrast, 16% of the Nandi children had no distinction was made between EBV-specific immunosup- positive blood smears after a recent malaria epidemic, dem- pression and generalized depression of T cell immunity. onstrating the dramatic difference in malaria exposure between The study described here investigated the effect of malaria the 2 populations. on EBV immunity and assessed whether the intensity and du- Sample collection. Peripheral blood was collected in so- ration of malaria exposure influences EBV-specific IFN-g and dium-heparinized tubes and transported to the Case Western IL-10 responses in healthy children from 2 epidemiologically Reserve University laboratory located at KEMRI’s Center for distinct areas of western Kenya that differ markedly in the Vector Biology and Control Research in Kisumu for processing incidence of eBL [3, 5, 27, 28]. In addition, malaria-specific the same day. Peripheral blood mononuclear cells (PBMCs) cytokine responses were examined concomitantly to address were separated from whole blood by ficoll-hypaque density the question of whether there are global versus EBV-specific gradient centrifugation and suspended in RPMI 1640 (GIBCO) alterations in T cell immunity in children exposed to malaria. supplemented with 10% heat-inactivated human AB serum, 50 Table 1. Study participant demographics. Age group Site, characteristic 1–4 years 5–9 years 10–14 years All Kisumu Study participants enrolled, no. 32 36 36 104 Hemoglobin level, mean, g/dL 9.7 12.2 12.5 11.6 Children with malaria-positive smear,a %77728377 Body temperature, mean, ؇C 36.9 36.7 37.0 36.9 Nandi Study participants enrolled, no. 36 48 43 127 Hemoglobin level, mean, g/dL 12.0 12.9 13.5 12.8 Children with malaria-positive smear,a % 8 14 26 16 Body temperature, mean, ؇C 37.2 37.3 37.0 37.2 a Plasmodium falciparum prevalence differed significantly between study sites (P ! .0001 ).
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