Human Genetics

, Volume 121, Issue 2, pp 223–232 | Cite as

PD-1 gene haplotype is associated with the development of type 1 diabetes mellitus in Japanese children

  • Ronghua Ni
  • Kenji IharaEmail author
  • Kenichi Miyako
  • Ryuichi Kuromaru
  • Mika Inuo
  • Hitoshi Kohno
  • Toshiro Hara
Original Investigation


Interaction between Programmed cell death-1 (PD-1), a member of costimulatory molecules, and its receptors Programmed cell Death-1 Ligand 1 (PD-L1) and Programmed cell Death-1 Ligand 2 (PD-L2), play an important role in the negative regulation of immune reactions. It was shown that a polymorphism in a regulatory site of the PD-1 gene was associated with susceptibility to several autoimmune diseases in various ethnic groups, whereas the contribution of the PD-1 gene or its ligand genes to the onset of type 1 diabetes (T1D) mellitus in the Japanese population remains unknown. We first screened PD-1, PD-L1, and PD-L2 genes for polymorphisms in the Japanese population, and then investigated the frequencies of polymorphisms in patients with T1D mellitus in comparison with healthy controls. In total, we identified 26 polymorphic sites within these genes, and then 23 polymorphisms with minor allele frequencies greater than 5% were intensively analyzed for genotyping in the patients and the controls. As a result, allele and genotype frequencies of the polymorphism numbers 2, 3, 4, 5, 6, and 8 in the PD-1 gene were different to some extent between the patients and the controls with P < 0.05, which did not reach statistical significance after the correction of multiple comparisons. Allele or genotype frequencies of any SNPs in the PD-L1 or PD-L2 gene did not show differences between the patients and the controls. The frequencies of the estimated haplotypes, those of which consisted of polymorphism numbers 2, 3, 4, 5, 6, and 8 in the PD-1, were significantly different between the patients and the controls (P = 0.00095). The in vitro assessment for a transcription activity of each haplotype of the PD-1 gene by luciferase assay did not demonstrate a functional difference between the haplotypes. In conclusion, the genetic evaluation by association study demonstrated that the PD-1 gene was a predisposing gene to the development of T1D mellitus in the Japanese population.


Costimulatory Pathway Estimate Haplotype Frequency ICOS Gene Rheumatoid Athritis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



Type 1 diabetes mellitus


Systemic lupus erythematosus


Dendritic cell


Restriction fragment length polymorphism


Single strand conformation polymorphism



This work was supported by a Grant-in Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.


  1. Ahmed S, Ihara K, Bassuny WM, Kuromaru R, Kohno H, Miyako K, Matsuura N, Iwata I, Nagafuchi S, Hara T (2002) Association study between CD30 and CD30 ligand genes and type 1 diabetes in the Japanese population. Genes Immun 3:96–101PubMedCrossRefGoogle Scholar
  2. Ansari MJ, Salama AD, Chitnis T, Smith RN, Yagita H, Akiba H, Yamazaki T, Azuma M, Iwai H, Khoury SJ, Auchincloss H Jr, Sayegh MH (2003) The programmed death-1 (PD-1) pathway regulates autoimmune diabetes in nonobese diabetic (NOD) mice. J Exp Med 198:63–69PubMedCrossRefGoogle Scholar
  3. Barker JM (2006) Clinical review: type 1 diabetes-associated autoimmunity: natural history, genetic associations, and screening. J Clin Endocrinol Metab 91:1210–1217PubMedCrossRefGoogle Scholar
  4. Bassuny WM, Ihara K, Sasaki Y, Kuromaru R, Kohno H, Matsuura N, Hara T (2003) A functional polymorphism in the promoter/enhancer region of the FOXP3/Scurfin gene associated with type 1 diabetes. Immunogenetics 55:149–156PubMedCrossRefGoogle Scholar
  5. Ferreiros-Vidal I, Gomez-Reino JJ, Barros F, Carracedo A, Carreira P, Gonzalez-Escribano F, Liz M, Martin J, Ordi J, Vicario JL, Gonzalez A (2004) Association of PDCD1 with susceptibility to systemic lupus erythematosus: evidence of population-specific effects. Arthritis Rheum 50:2590–2597PubMedCrossRefGoogle Scholar
  6. Freeman GJ, Long AJ, Iwai Y, Bourque K, Chernova T, Nishimura H, Fitz LJ, Malenkovich N, Okazaki T, Byrne MC, Horton HF, Fouser L, Carter L, Ling V, Bowman MR, Carreno BM, Collins M, Wood CR, Honjo T (2000) Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation. J Exp Med 192:1027–1034PubMedCrossRefGoogle Scholar
  7. Giles RH, Peters DJM, Breuning MH (1998) Conjunction dysfunction: CBP/p300 in human disease. Trends Genet 14:178PubMedCrossRefGoogle Scholar
  8. Gray CP, Arosio P, Hersey P (2002) Heavy chain ferritin activates regulatory T cells by induction of changes in dendritic cells. Blood 99:3326–3334PubMedCrossRefGoogle Scholar
  9. Hawa MI, Beyan H, Buckley LR, Leslie RD (2002) Impact of genetic and non-genetic factors in type 1 diabetes. Am J Med Genet 115:8–17PubMedCrossRefGoogle Scholar
  10. Johansson M, Arlestig L, Moller B, Rantapaa-Dahlqvist S (2005) Association of a PDCD1 polymorphism with renal manifestations in systemic lupus erythematosus. Arthritis Rheum 52:1665–1669PubMedCrossRefGoogle Scholar
  11. Karvonen M, Viik-Kajander M, Moltchanova E, Libman I, LaPorte R, Tuomilehto J (2000) Incidence of childhood type 1 diabetes worldwide. Diabetes mondiale (diamond) project group. Diabetes Care 23:1516–1526PubMedCrossRefGoogle Scholar
  12. Kong EK, Prokunina-Olsson L, Wong WH, Lau CS, Chan TM, Alarcon-Riquelme M, Lau YL (2005) A new haplotype of PDCD1 is associated with rheumatoid arthritis in Hong Kong Chinese. Arthritis Rheum 52:1058–1062PubMedCrossRefGoogle Scholar
  13. Kroner A, Mehling M, Hemmer B, Rieckmann P, Toyka KV, Maurer M, Wiendl H (2005) A PD-1 polymorphism is associated with disease progression in multiple sclerosis. Ann Neurol 58:50–57PubMedCrossRefGoogle Scholar
  14. Latchman Y, Wood CR, Chernova T, Chaudhary D, Borde M, Chernova I, Iwai Y, Long AJ, Brown JA, Nunes R, Greenfield EA, Bourque K, Boussiotis VA, Carter LL, Carreno BM, Malenkovich N, Nishimura H, Okazaki T, Honjo T, Sharpe AH, Freeman GJ (2001) PD-L2 is a second ligand for PD-1 and inhibits T cell activation. Nat Immunol 2:261–268PubMedCrossRefGoogle Scholar
  15. Lewontin RC (1995) The detection of linkage disequilibrium in molecular sequence data. Genetics 140:377–388PubMedGoogle Scholar
  16. Lin SC, Yen JH, Tsai JJ, Tsai WC, Ou TT, Liu HW, Chen CJ (2004) Association of a programmed death 1 gene polymorphism with the development of rheumatoid arthritis, but not systemic lupus erythematosus. Arthritis Rheum 50:770–775PubMedCrossRefGoogle Scholar
  17. Marron MP, Zeidler A, Raffel LJ, Eckenrode SE, Yang JJ, Hopkins DI, Garchon HJ, Jacob CO, Serrano-Rios M, Martinez Larrad MT, Park Y, Bach JF, Rotter JI, Yang MC, She JX (2000) Genetic and physical mapping of a type 1 diabetes susceptibility gene (IDDM12) to a 100-kb phagemid artificial chromosome clone containing D2S72-CTLA4-D2S105 on chromosome 2q33. Diabetes 49:492–499PubMedCrossRefGoogle Scholar
  18. Matsuura N, Fukuda K, Okuno A, Harada S, Fukushima N, Koike A, Ito Y, Hotsubo T (1998) Descriptive epidemiology of IDDM in Hokkaido, Japan: the childhood IDDM Hokkaido registry. Diabetes Care 21:1632–1636PubMedCrossRefGoogle Scholar
  19. Nielsen C, Hansen D, Husby S, Jacobsen BB, Lillevang ST (2003) Association of a putative regulatory polymorphism in the PD-1 gene with susceptibility to type 1 diabetes. Tissue Antigens 62:492–497PubMedCrossRefGoogle Scholar
  20. Nielsen C, Laustrup H, Voss A, Junker P, Husby S, Lillevang ST (2004) A putative regulatory polymorphism in PD-1 is associated with nephropathy in a population-based cohort of systemic lupus erythematosus patients. Lupus 13:510–516PubMedCrossRefGoogle Scholar
  21. Nishimura H, Nose M, Hiai H, Minato N, Honjo T (1999) Development of lupus-like autoimmune diseases by disruption of the PD-1 gene encoding an ITIM motif-carrying immunoreceptor. Immunity 11:141–151PubMedCrossRefGoogle Scholar
  22. Nishimura H, Okazaki T, Tanaka Y, Nakatani K, Hara M, Matsumori A, Sasayama S, Mizoguchi A, Hiai H, Minato N, Honjo T (2001) Autoimmune dilated cardiomyopathy in PD-1 receptor-deficient mice. Science 291:319–322PubMedCrossRefGoogle Scholar
  23. Okazaki T, Maeda A, Nishimura H, Kurosaki T, Honjo T (2001) PD-1 immunoreceptor inhibits B cell receptor-mediated signaling by recruiting src homology 2-domain-containing tyrosine phosphatase 2 to phosphotyrosine. Proc Natl Acad Sci USA 98:13866–13871PubMedCrossRefGoogle Scholar
  24. Okazaki T, Tanaka Y, Nishio R, Mitsuiye T, Mizoguchi A, Wang J, Ishida M, Hiai H, Matsumori A, Minato N, Honjo T (2003) Autoantibodies against cardiac troponin I are responsible for dilated cardiomyopathy in PD-1-deficient mice. Nat Med 9:1477–1483PubMedCrossRefGoogle Scholar
  25. Okazaki T, Wang J (2005) PD-1/PD-L pathway and autoimmunity. Autoimmunity 38:353–357PubMedCrossRefGoogle Scholar
  26. Park MY, Jang HD, Lee SY, Lee KJ, Kim E (2004) Fas-associated factor-1 inhibits nuclear factor-kappaB (NF-kappaB) activity by interfering with nuclear translocation of the RelA (p65) subunit of NF-kappaB. J Biol Chem 279:2544–2549PubMedCrossRefGoogle Scholar
  27. Pociot F, McDermott MF (2002) Genetics of type 1 diabetes mellitus. Genes Immun 3:235–249PubMedCrossRefGoogle Scholar
  28. Prokunina L, Castillejo-Lopez C, Oberg F, Gunnarsson I, Berg L, Magnusson V, Brookes AJ, Tentler D, Kristjansdottir H, Grondal G, Bolstad AI, Svenungsson E, Lundberg I, Sturfelt G, Jonssen A, Truedsson L, Lima G, Alcocer-Varela J, Jonsson R, Gyllensten UB, Harley JB, Alarcon-Segovia D, Steinsson K, Alarcon-Riquelme ME (2002) A regulatory polymorphism in PDCD1 is associated with susceptibility to systemic lupus erythematosus in humans. Nat Genet 32:666–669PubMedCrossRefGoogle Scholar
  29. Prokunina L, Padyukov L, Bennet A, de Faire U, Wiman B, Prince J, Alfredsson L, Klareskog L, Alarcon-Riquelme M (2004) Association of the PD-1.3A allele of the PDCD1 gene in patients with rheumatoid arthritis negative for rheumatoid factor and the shared epitope. Arthritis Rheum 50:1770–1773PubMedCrossRefGoogle Scholar
  30. Reddy MV, Johansson M, Sturfelt G, Jonsen A, Gunnarsson I, Svenungsson E, Rantapaa-Dahlqvist S, Alarcon-Riquelme ME (2005) The R620W C/T polymorphism of the gene PTPN22 is associated with SLE independently of the association of PDCD1. Genes Immun 6:658–662PubMedGoogle Scholar
  31. Salama AD, Chitnis T, Imitola J, Ansari MJ, Akiba H, Tushima F, Azuma M, Yagita H, Sayegh MH, Khoury SJ (2003) Critical role of the programmed death-1 (PD-1) pathway in regulation of experimental autoimmune encephalomyelitis. J Exp Med 198:71–78PubMedCrossRefGoogle Scholar
  32. Sanghera DK, Manzi S, Bontempo F, Nestlerode C, Kamboh MI (2004) Role of an intronic polymorphism in the PDCD1 gene with the risk of sporadic systemic lupus erythematosus and the occurrence of antiphospholipid antibodies. Hum Genet 115:393–398PubMedCrossRefGoogle Scholar
  33. Sasaki Y, Ihara K, Matsuura N, Kohno H, Nagafuchi S, Kuromaru R, Kusuhara K, Takeya R, Hoey T, Sumimoto H, Hara T (2004) Identification of a novel type 1 diabetes susceptibility gene, T-bet. Hum Genet 115:177–184PubMedCrossRefGoogle Scholar
  34. The Expert Committee on the Diagnosis, Classification of Diabetes Mellitus (1997) Report of the expert committee on the diagnosis and classification of diabetes mellitus. Diabetes Care 20:1183–1197Google Scholar
  35. Tisch R, McDevitt H (1996) Insulin-dependent diabetes mellitus. Cell 85:291–297PubMedCrossRefGoogle Scholar
  36. Tseng SY, Otsuji M, Gorski K, Huang X, Slansky JE, Pai SI, Shalabi A, Shin T, Pardoll DM, Tsuchiya H (2001) B7-DC, a new dendritic cell molecule with potent costimulatory properties for T cells. J Exp Med 193:839–846PubMedCrossRefGoogle Scholar
  37. Tsutsumi Y, Jie X, Ihara K, Nomura A, Kanemitsu S, Takada H, Hara T (2006) Phenotypic and genetic analyses of T cell-mediated immunoregulation in patients with type 1 diabetes. Diabet Med 23:1145–1150PubMedCrossRefGoogle Scholar
  38. Wang J, Yoshida T, Nakaki F, Hiai H, Okazaki T, Honjo T (2005) Establishment of NOD-Pdcd1-/- mice as an efficient animal model of type I diabetes. Proc Natl Acad Sci USA 102:11823–11828PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Ronghua Ni
    • 1
  • Kenji Ihara
    • 1
    Email author
  • Kenichi Miyako
    • 1
  • Ryuichi Kuromaru
    • 1
  • Mika Inuo
    • 1
  • Hitoshi Kohno
    • 2
  • Toshiro Hara
    • 1
  1. 1.Department of Pediatrics, Graduate School of Medical SciencesKyushu UniversityFukuokaJapan
  2. 2.Department of Endocrinology and MetabolismFukuoka Children’s HospitalFukuokaJapan

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