Human Genetics

, Volume 124, Issue 5, pp 515–524 | Cite as

Haplotypes of IL-10 promoter variants are associated with susceptibility to severe malarial anemia and functional changes in IL-10 production

  • Collins Ouma
  • Gregory C. Davenport
  • Tom Were
  • Michael F. Otieno
  • James B. Hittner
  • John M. Vulule
  • Jeremy Martinson
  • John M. Ong’echa
  • Robert E. Ferrell
  • Douglas J. Perkins
Original Investigation


Plasmodium falciparum malaria is one of the leading global causes of morbidity and mortality with African children bearing the highest disease burden. Among the various severe disease sequelae common to falciparum malaria, severe malarial anemia (SMA) in pediatric populations accounts for the greatest degree of mortality. Although the patho-physiological basis of SMA remains unclear, dysregulation in inflammatory mediators, such as interleukin (IL)-10, appear to play an important role in determining disease outcomes. Since polymorphic variability in innate immune response genes conditions susceptibility to malaria, the relationship between common IL-10 promoter variants (−1,082A/G, −819T/C, and −592A/C), SMA (Hb < 6.0 g/dL), and circulating inflammatory mediator levels (i.e., IL-10, TNF-α, IL-6 and IL-12) were investigated in parasitemic Kenyan children (n = 375) in a holoendemic P. falciparum transmission area. Multivariate logistic regression analyses demonstrated that the −1,082G/−819C/−592C (GCC) haplotype was associated with protection against SMA (OR; 0.68, 95% CI, 0.43–1.05; = 0.044) and increased IL-10 production (= 0.029). Although none of the other haplotypes were significantly associated with susceptibility to SMA, individuals with the −1,082A/−819T/−592A (ATA) haplotype had an increased risk of SMA and reduced circulating IL-10 levels (= 0.042). Additional results revealed that the IL-10:TNF-α ratio was higher in the GCC group (= 0.024) and lower in individuals with the ATA haplotype (= 0.034), while the IL-10:IL-12 ratio was higher in ATA haplotype (= 0.006). Results presented here demonstrate that common IL-10 promoter haplotypes condition susceptibility to SMA and functional changes in circulating IL-10, TNF-α, and IL-12 levels in children with falciparum malaria.


  1. Aidoo M, Terlouw DJ, Kolczak MS, McElroy PD, ter Kuile FO, Kariuki S, Nahlen BL, Lal AA, Udhayakumar V (2002) Protective effects of the sickle cell gene against malaria morbidity and mortality. Lancet 359:1311–1312PubMedCrossRefGoogle Scholar
  2. Awandare GA, Ouma C, Keller CC, Were T, Otieno R, Ouma Y, Davenport GC, Hittner JB, Ong’echa JM, Ferrell R, Perkins DJ (2006) A macrophage migration inhibitory factor promoter polymorphism is associated with high-density parasitemia in children with malaria. Genes Immun 7:568–575PubMedCrossRefGoogle Scholar
  3. Berkley J, Mwarumba S, Bramham K, Lowe B, Marsh K (1999) Bacteraemia complicating severe malaria in children. Trans R Soc Trop Med Hyg 93:283–286PubMedCrossRefGoogle Scholar
  4. Bloland PB, Boriga DA, Ruebush TK, McCormick JB, Roberts JM, Oloo AJ, Hawley W, Lal A, Nahlen B, Campbell CC (1999) Longitudinal cohort study of the epidemiology of malaria infections in an area of intense malaria transmission II. Descriptive epidemiology of malaria infection and disease among children. Am J Trop Med Hyg 60:641–648PubMedGoogle Scholar
  5. Breman JG, Egan A, Keusch GT (2001) The intolerable burden of malaria: a new look at the numbers. Am J Trop Med Hyg 64:4–7Google Scholar
  6. Burgner D, Usen S, Rockett K, Jallow M, Ackerman H, Cervino A, Pinder M, Kwiatkowski DP (2003) Nucleotide and haplotypic diversity of the NOS2A promoter region and its relationship to cerebral malaria. Hum Genet 112:379–386PubMedGoogle Scholar
  7. Cai G, Kastelein RA, Hunter CA (1999) IL-10 enhances NK cell proliferation, cytotoxicity and production of IFN-gamma when combined with IL-18. Eur J Immunol 29:2658–2665PubMedCrossRefGoogle Scholar
  8. Carpenter D, Abushama H, Bereczky S, Farnert A, Rooth I, Troye-Blomberg M, Quinnell RJ, Shaw MA (2007) Immunogenetic control of antibody responsiveness in a malaria endemic area. Hum Immunol 68:165–169PubMedCrossRefGoogle Scholar
  9. Clark IA, Chaudhri G (1988) Tumour necrosis factor may contribute to the anaemia of malaria by causing dyserythropoiesis and erythrophagocytosis. Br J Haematol 70:99–103PubMedCrossRefGoogle Scholar
  10. Conti P, Kempuraj D, Kandere K, Di Gioacchino M, Barbacane RC, Castellani ML, Felaco M, Boucher W, Letourneau R, Theoharides TC (2003) IL-10, an inflammatory/inhibitory cytokine, but not always. Immunol Lett 86:123–129PubMedCrossRefGoogle Scholar
  11. Cramer JP, Mockenhaupt FP, Ehrhardt S, Burkhardt J, Otchwemah RN, Dietz E, Gellert S, Bienzle U (2004) iNOS promoter variants and severe malaria in Ghanaian children. Trop Med Int Health 9:1074–1080PubMedCrossRefGoogle Scholar
  12. Crawley E, Kay R, Sillibourne J, Patel P, Hutchinson I, Woo P (1999) Polymorphic haplotypes of the interleukin-10 5′ flanking region determine variable interleukin-10 transcription and are associated with particular phenotypes of juvenile rheumatoid arthritis. Arthritis Rheum 42:1101–1108PubMedCrossRefGoogle Scholar
  13. Eskdale J, Gallagher G (1995) A polymorphic dinucleotide repeat in the human IL-10 promoter. Immunogenetics 42:444–445PubMedCrossRefGoogle Scholar
  14. Eskdale J, Gallagher G, Verweij CL, Keijsers V, Westendorp RG, Huizinga TW (1998) Interleukin 10 secretion in relation to human IL-10 locus haplotypes. Proc Natl Acad Sci USA 95:9465–9470PubMedCrossRefGoogle Scholar
  15. Fiorentino DF, Zlotnik A, Mosmann TR, Howard M, O’Garra A (1991) IL-10 inhibits cytokine production by activated macrophages. J Immunol 147:3815–3822PubMedGoogle Scholar
  16. Gibson AW, Edberg JC, Wu J, Westendorp RG, Huizinga TW, Kimberly RP (2001) Novel single nucleotide polymorphisms in the distal IL-10 promoter affect IL-10 production and enhance the risk of systemic lupus erythematosus. J Immunol 166:3915–3922PubMedGoogle Scholar
  17. Giordani L, Bruzzi P, Lasalandra C, Quaranta M, Schittulli F, Della Ragione F, Iolascon A (2003) Association of breast cancer and polymorphisms of interleukin-10 and tumor necrosis factor-alpha genes. Clin Chem 49:1664–1667PubMedCrossRefGoogle Scholar
  18. Hobbs MR, Udhayakumar V, Levesque MC, Booth J, Roberts JM, Tkachuk AN, Pole A, Coon H, Kariuki S, Nahlen BL, Mwaikambo ED, Lal AL, Granger DL, Anstey NM, Weinberg JB (2002) A new NOS2 promoter polymorphism associated with increased nitric oxide production and protection from severe malaria in Tanzanian and Kenyan children. Lancet 360:1468–1475PubMedCrossRefGoogle Scholar
  19. Ide A, Kawasaki E, Abiru N, Sun F, Takahashi R, Kuwahara H, Fujita N, Kita A, Oshima K, Sakamaki H, Uotani S, Yamasaki H, Yamaguchi Y, Eguchi K (2002) Genetic association between interleukin-10 gene promoter region polymorphisms and type 1 diabetes age-at-onset. Hum Immunol 63:690–695PubMedCrossRefGoogle Scholar
  20. Jason J, Archibald LK, Nwanyanwu OC, Bell M, Buchanan I, Larned J, Kazembe PN, Dobbie H, Parekh B, Byrd MG, Eick A, Han A, Jarvis WR (2001) Cytokines and malaria parasitemia. Clin Immunol 100:208–218PubMedCrossRefGoogle Scholar
  21. Keijsers V, Verweij CL, Westendorp RG, Breedveld FC, Huizinga T (1997) IL-10 polymorphisms in relation to production and rheumatoid arthritis. Arthritis Rheum 43:120Google Scholar
  22. Keller CC, Yamo O, Ouma C, Ong’echa JM, Ounah D, Hittner JB, Vulule JM, Perkins DJ (2006) Acquisition of hemozoin by monocytes down-regulates interleukin-12 p40 (IL-12p40) transcripts and circulating IL-12p70 through an IL-10-dependent mechanism: in vivo and in vitro findings in severe malarial anemia. Infect Immun 74:5249–5260PubMedCrossRefGoogle Scholar
  23. Kube D, Platzer C, von Knethen A, Straub H, Bohlen H, Hafner M, Tesch H (1995) Isolation of the human interleukin 10 promoter. Characterization of the promoter activity in Burkitt’s lymphoma cell lines. Cytokine 7:1–7PubMedCrossRefGoogle Scholar
  24. Kurtis JD, Lanar DE, Opollo M, Duffy PE (1999) Interleukin-10 responses to liver-stage antigen 1 predict human resistance to Plasmodium falciparum. Infect Immun 67:3424–3429PubMedGoogle Scholar
  25. Kurtzhals JA, Akanmori BD, Goka BQ, Adabayeri V, Nkrumah FK, Behr C, Hviid L (1999) The cytokine balance in severe malarial anemia. J Infect Dis 180:1753–1755PubMedCrossRefGoogle Scholar
  26. Kwiatkowski DP (2005) How malaria has affected the human genome and what human genetics can teach us about malaria. Am J Hum Genet 77:171–192PubMedCrossRefGoogle Scholar
  27. Lalani I, Bhol K, Ahmed AR (1997) Interleukin-10: biology, role in inflammation and autoimmunity. Ann Allergy Asthma Immunol 79:469–483PubMedCrossRefGoogle Scholar
  28. Lee JY, Kim HY, Kim KH, Kim SM, Jang MK, Park JY, Lee JH, Kim JH, Yoo JY (2005) Association of polymorphism of IL-10 and TNF-A genes with gastric cancer in Korea. Cancer Lett 225:207–214PubMedCrossRefGoogle Scholar
  29. Lim S, Crawley E, Woo P, Barnes PJ (1998) Haplotype associated with low interleukin-10 production in patients with severe asthma. Lancet 352:113PubMedCrossRefGoogle Scholar
  30. Linke A, Kuhn R, Muller W, Honarvar N, Li C, Langhorne J (1996) Plasmodium chabaudi chabaudi: differential susceptibility of gene-targeted mice deficient in IL-10 to an erythrocytic-stage infection. Exp Parasitol 84:253–263PubMedCrossRefGoogle Scholar
  31. Luty AJ, Perkins DJ, Lell B, Schmidt-Ott R, Lehman LG, Luckner D, Greve B, Matousek P, Herbich K, Schmid D, Weinberg JB, Kremsner PG (2000) Low interleukin-12 activity in severe Plasmodium falciparum malaria. Infect Immun 68:3909–3915PubMedCrossRefGoogle Scholar
  32. Mangia A, Santoro R, Piattelli M, Pazienza V, Grifa G, Iacobellis A, Andriulli A (2004) IL-10 haplotypes as possible predictors of spontaneous clearance of HCV infection. Cytokine 25:103–109PubMedCrossRefGoogle Scholar
  33. May J, Lell B, Luty AJ, Meyer CG, Kremsner PG (2000) Plasma interleukin-10:Tumor necrosis factor (TNF)-alpha ratio is associated with TNF promoter variants and predicts malarial complications. J Infect Dis 182:1570–1573PubMedCrossRefGoogle Scholar
  34. McDevitt MA, Xie J, Gordeuk V, Bucala R (2004) The anemia of malaria infection: role of inflammatory cytokines. Curr Hematol Rep 3:97–106PubMedGoogle Scholar
  35. McElroy PD, Lal AA, Hawley WA, Bloland PB, Kuile FO, Oloo AJ, Harlow SD, Lin X, Nahlen BL (1999) Analysis of repeated hemoglobin measures in full-term, normal birth weight Kenyan children between birth and four years of age. III. The Asembo Bay Cohort Project. Am J Trop Med Hyg 61:932–940PubMedGoogle Scholar
  36. Meenagh A, Williams F, Ross OA, Patterson C, Gorodezky C, Hammond M, Leheny WA, Middleton D (2002) Frequency of cytokine polymorphisms in populations from western Europe, Africa, Asia, the Middle East and South America. Hum Immunol 63:1055–1061PubMedCrossRefGoogle Scholar
  37. Miller LH, Good MF, Milon G (1994) Malaria pathogenesis. Science 264:1878–1883PubMedCrossRefGoogle Scholar
  38. Ong’echa JM, Keller CC, Were T, Ouma C, Otieno RO, Landis-Lewis Z, Ochiel D, Slingluff JL, Mogere S, Ogonji GA, Orago AS, Vulule JM, Kaplan SS, Day RD, Perkins DJ (2006) Parasitemia, anemia, and malarial anemia in infants and young children in a rural holoendemic Plasmodium falciparum transmission area. Am J Trop Med Hyg 74:376–385PubMedGoogle Scholar
  39. Ong’echa JM, Remo AM, Kristoff J, Hittner JB, Were T, Ouma C, Otieno RO, Vulule JM, Keller CC, Awandare GA, Perkins DJ (2008) Increased circulating interleukin (IL)-23 in children with malarial anemia: in vivo and in vitro relationship with co-regulatory cytokines IL-12 and IL-10. Clin Immunol 126:211–221PubMedCrossRefGoogle Scholar
  40. Othoro C, Lal AA, Nahlen B, Koech D, Orago AS, Udhayakumar V (1999) A low interleukin-10 tumor necrosis factor-alpha ratio is associated with malaria anemia in children residing in a holoendemic malaria region in western Kenya. J Infect Dis 179:279–282PubMedCrossRefGoogle Scholar
  41. Otieno RO, Ouma C, Ong’echa JM, Keller CC, Were T, Waindi EN, Michaels MG, Day RD, Vulule JM, Perkins DJ (2006) Increased severe anemia in HIV-1-exposed and HIV-1-positive infants and children during acute malaria. Aids 20:275–280PubMedCrossRefGoogle Scholar
  42. Ouma C, Keller CC, Opondo DA, Were T, Otieno RO, Otieno MF, Orago AS, Ong’Echa JM, Vulule JM, Ferrell RE, Perkins DJ (2006) Association of Fc gamma receptor IIA (CD32) polymorphism with malarial anemia and high-density parasitemia in infants and young children. Am J Trop Med Hyg 74:573–577PubMedGoogle Scholar
  43. Ouma C, Davenport GC, Awandare GA, Keller CC, Were T, Otieno MF, Vulule JM, Martinson J, Ong’echa JM, Ferrell RE, Perkins DJ (2008) Polymorphic variability in the interleukin (IL)-1beta promoter conditions susceptibility to severe malarial anemia and functional changes in IL-1beta production. J Infect Dis 198:1219–1226PubMedCrossRefGoogle Scholar
  44. Perkins DJ, Weinberg JB, Kremsner PG (2000) Reduced interleukin-12 and transforming growth factor-beta1 in severe childhood malaria: relationship of cytokine balance with disease severity. J Infect Dis 182:988–992PubMedCrossRefGoogle Scholar
  45. Prakash D, Fesel C, Jain R, Cazenave PA, Mishra GC, Pied S (2006) Clusters of cytokines determine malaria severity in Plasmodium falciparum-infected patients from endemic areas of Central India. J Infect Dis 194:198–207PubMedCrossRefGoogle Scholar
  46. Scassellati C, Zanardini R, Squitti R, Bocchio-Chiavetto L, Bonvicini C, Binetti G, Zanetti O, Cassetta E, Gennarelli M (2004) Promoter haplotypes of interleukin-10 gene and sporadic Alzheimer’s disease. Neurosci Lett 356:119–122PubMedCrossRefGoogle Scholar
  47. Snow RW, Craig MH, Deichmann U, le Sueur D (1999) A preliminary continental risk map for malaria mortality among African children. Parasitol Today 15:99–104PubMedCrossRefGoogle Scholar
  48. Turner DM, Williams DM, Sankaran D, Lazarus M, Sinnott PJ, Hutchinson IV (1997) An investigation of polymorphism in the interleukin-10 gene promoter. Eur J Immunogenet 24:1–8PubMedGoogle Scholar
  49. Upperman JS, Pillage G, Siddiqi MQ, Zeevi A, Kelly N, Ford HR, Kammerer C, Spolarics Z (2005) Dominance of high-producing interleukin 6 and low-producing interleukin 10 and interferon gamma alleles in glucose-6-phosphate dehydrogenase-deficient trauma patients. Shock 23:197–201PubMedGoogle Scholar
  50. Wilson JN, Rockett K, Jallow M, Pinder M, Sisay-Joof F, Newport M, Newton J, Kwiatkowski D (2005) Analysis of IL10 haplotypic associations with severe malaria. Genes Immun 6:462–466PubMedCrossRefGoogle Scholar
  51. Winkler S, Willheim M, Baier K, Schmid D, Aichelburg A, Graninger W, Kremsner PG (1998) Reciprocal regulation of Th1- and Th2-cytokine-producing T cells during clearance of parasitemia in Plasmodium falciparum malaria. Infect Immun 66:6040–6044PubMedGoogle Scholar
  52. WHO (2000) Severe falciparum malaria. Trans R Soc Trop Med Hyg 94(Supplement 1):S1–S90Google Scholar
  53. WHO (2005) World Malaria Report 2005. Geneva: World Health Organization/United Nations Children’s Fund; 2005:
  54. Wunderink RG, Waterer GW (2003) Genetics of sepsis and pneumonia. Curr Opin Crit Care 9:384–389PubMedCrossRefGoogle Scholar
  55. Yao JG, Gao LB, Liu YG, Li J, Pang GF (2008) Genetic variation in interleukin-10 gene and risk of oral cancer. Clin Chim Acta 388:84–88PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Collins Ouma
    • 1
  • Gregory C. Davenport
    • 2
  • Tom Were
    • 1
    • 3
  • Michael F. Otieno
    • 4
  • James B. Hittner
    • 5
  • John M. Vulule
    • 1
  • Jeremy Martinson
    • 2
  • John M. Ong’echa
    • 1
    • 6
  • Robert E. Ferrell
    • 7
  • Douglas J. Perkins
    • 1
    • 6
  1. 1.Centre for Global Health ResearchKenya Medical Research Institute, University of New Mexico/KEMRI Laboratories of Parasitic and Viral DiseasesKisumuKenya
  2. 2.Department of Infectious Diseases and Microbiology, Graduate School of Public HealthUniversity of PittsburghPittsburghUSA
  3. 3.Department of PathologyKenyatta UniversityNairobiKenya
  4. 4.Department of Pre-Clinical SciencesKenyatta UniversityNairobiKenya
  5. 5.Department of PsychologyCollege of CharlestonCharlestonUSA
  6. 6.Division of Infectious DiseasesUniversity of New Mexico School of MedicineAlbuquerqueUSA
  7. 7.Department of Human Genetics, Graduate School of Public HealthUniversity of PittsburghPittsburghUSA

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