Kekkaku Vol. 85, No. 6: 501_508, 2010 501 ミニ特集 免疫と結核 要旨 3 1 20 940 180 WHO 2008 1) 11) 1998 1999 T Th1 T Mφ DBA/1 BALB/c C57BL/6 DBA/1 BALB/c T 1 T 2 CpG TLR9 CpG 3 Lipocalin 2 SLPI 4 T granulysin HSP 65+IL-12 DNA 5 キーワーズ T Ⅰ. はじめに T Th1 T Mφ Mφ _ T Ⅱ. 結核症 1 HIV 2007 10 1.7 10 19.8 5 5 10% 3 1 dormancy TNFα Ⅲ. 獲得免疫と結核 Mφ CD4 + T NK γ/δt T CD8 + T CD8 T Fig. 1 1998 Nature H37Rv 1 T CD8 + T 591 _ 8555 1180 E-mail : okm@kch.hosp.go.jp Received 21 Mar. 2010
502 85 6 2010 6 CD8 + T Fig. 2 T granulysin MHC 38 kda HSP65 CD8 + T 19 kda Ag85 CFP10 Mtb11 CD8 + T 10) ESAT-6 T HLA-A2 82 90 9 AMASTEGNV T SCID-PBL/hu ESAT-6 HLA-A2 T 2) 12 2 T CD4 + T MHC Class (-/-) CD4 (-/-) CD4 Th1 Killing of M.tuberculosis M.tuberculosis Mφ granulysin IL-18 IL-12 Suppression NK cell perforin TNF-α IL-1 IL-2 IFN-γ IL-10 IL-4 IFN-γ CTL Th1 cell Th0 cell Differentiation DNA vaccine Differentiation granuloma formation Mφ activation inos induction Fig. 1 Mφ and T cell immunity against tuberculosis 1 Ⅳ. 自然免疫と結核 1 Mφ Mφ Mφ 2) 3) ROI NO RNI TACO Nramp 2 Toll-like Pathogen Recognition Receptor Toll-like receptor TLR innate immunity 13) TLR TLR1 TLR10 3 Fig. 3A TLR TLR1 TLR2 TLR4 TLR6 TLR9 cell wall LAM magp total lipid TLR2 Fig. 3B TLR2 TLR4 M. tuberculosis Man LAM Mφ glycolipid Ara LAM TLR2 Mφ 19kDa lipoprotein TLR2 Mφ DNA CpG CpG TLR9 TLR2 2 Arg753 Gln Arg677 Trp Arg753 Gln CTL Differentiation IL-2 IL-6 granulysin Fas ligand TRAIL Fas TRAIL R M. TB Fungus Bacteria early late Apoptosis Precursor of CTL IL-5 IFN-γ IL-1, 7, 12, 13, 15, 18, 23, 27 IFN-α/β, TNF-α Effector CTL Perforin Granzyme A, B Destruction of tumor cell Destruction of virus infected cells Destruction of bacteria infected cells (Mφ) Fig. 2 Induction of CTL and granulysin
Immunity against MTB (Introduction) /M.Okada 503 MALP-2 BLP RNA flagellin R-848 CpG DNA LPS TLR2 TLR6 TLR1 TLR2 TLR3 TLR5 TLR7 TLR9 TLR4 TIR TIR TIR TIR TIR TIR TIR TIR TIR MyD88 TRAF6 IRAK-1/IRAK-4 IKKβ/NEMO TIRAP NF-κB NF-κB IRF-3 MyD88 IFN-β IFN MyD88 Fig. 3A TLR and pathogens Arg677 Trp M.leprae TLR TLR MyD88 MyD88 MyD88 MyD88 2 TLR TLR3 TLR4 IFN IFN MyD88 TRIF TRIF TLR3 TLR4 MyD88 TRAM MyD88 TRIF TLR4 TIRAP TLR MyD88 TLR1/2/6 TLR4 TRIF (-/-) MyD88 (-/-) TLR PRR (pathogen recognition receptor) DC-SIGN NOD dectin-1 HIV M.tuberculosis DC-SIGN TLR NOD1 NOD2 CARD Fig. 3B TLR LAM CWS peptidoglycan 19 kda lipoprotein CpG repeat TLR2 TLR2 /4 TLR2 /4 TLR2 TLR9 NOD2 PGN MDP 8) 3 RNA RIG-I, MDA5, LGP I TLR-3, -4, -7 RNA RNA DNA IFN TLR RNA RIG-I retinoic acid inducible gene I MDA5 melanoma differentiation associated gene 5 RNA RNA 5 -triphosphate IFN 14) 4 RNA RIG- I, MDA5, LGP2 RIG-I DExD/boxRNA
504 85 6 2010 6 Promiscuous sensing Immunogenic DNA or RNA HMGBs Discriminative sensing Cytosolic sensors RIG-I, MDA5 (RNA/DNA) DAI, AIM2, others (DNA) Nucleic-acid-sensing TLRs TLR9 (DNA) TLR3, TLR7 (RNA) Innate immune responses Fig. 3C Intracellular DNA sensor 14) RIG-I C RNA N 2 CARD IFN MDA5 RIG-I N 2 CARD C RIG-I RNA polyinosine-polycytidylic acid ploy I : C MDA5 LPG2 HVJ- RIG-I Fig. 3C 5 DNA DAI, AIM2, High Mobility Group Box HMGB DNA DAI DNA-dependent activator of IRFs AIM2 absent in melanoma 2 RLRs RIG -I like receptor DNA HMGB HMGB1, HMGB2 HMGB3 HMGB TLR3, TLR7, TLR9 HMGB TLR3, 7, 9, RIG-I, MDA5, DAI, AIM2 DNA Fig. 3C 15) 6 1 dendritic cell DC IFN DC DC pdc IFN-α DC pdc DC cdc 2 cdc GM-CSF pdc Flt-3-L TLR7 TLR9 Ⅴ. 結核菌抵抗性と宿主免疫 1 C57BL/6 BALB/c C57BL/6 T Th1 BALB/c T Th2 C57BL/6 Th1 BALB/c Th1 200 T DBA /1 BALB/c Fig. 4, 5, 6 DBA /1 BALB/c C57BL/6 PPD 5μg/ml 20μg/ml H37Ra 20μg/ml 100μg/ml in vitro 5 HSP65 T 51 Cr DBA /1 BALB/c T C57BL/6 T Fig. 5A T BALB/c T DBA/1 T Fig. 4 IFN-γ DBA /1 BALB/c IFNγ Fig. 6 IL-2 BALB/c C57BL/6 DBA /1
Immunity against MTB (Introduction) /M.Okada 505 BALB/c mouse DBA/1 mouse (%) (%) 30 30 % Specific cytotoxicity 20 20 10 10 stimulated with 0 0 ( ) EL-4 MMC IL-2 EL-4 MMC +IL-2 ( ) EL-4 MMC IL-2 EL-4 MMC +IL-2 Fig. 4 Comparison of CTL induction in vitro against M.tuberculosis antigens between DBA/1 mouse and BALB/c mouse (Target EL-4) (%) 60 C57BL/6 mice Target : P815 transfected with HSP65 DNA DBA/1 mice Target : DBA/1 Mφ transfected with HSP65 DNA 60 60 BALB/c mice Target : P815 transfected with HSP65 DNA % Specific cytotoxicity 40 20 40 40 20 20 stimulated with 0 ( ) PPD 5μg 20μg killed H37Ra 20μg 100μg 0 ( ) PPD 0 killed H37Ra 5μg 20μg 20μg 100μg ( ) PPD 5μg 20μg killed H37Ra 20μg 100μg Fig. 5A Comparison of CTL induction in vitro against M.tuberculosis antigens among DBA /1, BALB/c and C57BL /6 mice T Fig. 5B Ⅵ. サイトカインと結核 1 T T CD8 + T Tc T Th T Th CD4 + CD 8 T T CD8 + IL-2 IL-2 T Fig. 2 16) 18) IL-2 T T T IL-2 Th IL-6 IFN-γ T T 15) 16)
506 85 6 2010 6 Fig. 5B mouse strain strain 5 105 i.v. T IFN-γ C57BL/6 BALB/c DBA/1 + ++ ++++ >300 250 25 ++++ + ++++ + +++ (pg/ml) IFN-γ ELISA (U/ml) 3000 120 IL-2 ELISA 2500 100 2000 80 DBA/1 Balb/c C57BL/6 1500 60 1000 40 500 20 0 0 ( ) PPD TB 5μg 20μg 5μg 20μg 100μg PPD 5μg 20μg TB 5μg 20μg 100μg Fig. 6 IFN-γ and IL-2 productions from DBA/1, Balb/c or C57BL/6 mouse spleen cells IL-6 Tc 16) PBL T IL-2 IFN-γ IL-6 8) PPD T 8) 2 Mφ Mφ IFN-γ TNF-α IL-6 IL-12 3 IFN-γ TNF-α IL-6 IL-12 IL-15 Mφ IL-12 IL-18 TNF-α IL-1 T NK γδ T IFN-γ IFN-γ Mφ T T IL-12 IFN-γ IFN-γ Mφ IL-12 IL-12 T IFN-γ Mφ 3) IL-12 IL-18 IFN-γ αβ T Th1 IL-6 TNF-α Th1 Th1 dendritic cells : DC Mφ T IFN-γ DC T class CD4 + Th1 T class CD8 + T CD8 + T IFN-γ Mφ Mφ IL-15 T IL-10 Mφ IL-10, IL-19 IL-22 IL-28 IL-29 IL-10
Immunity against MTB (Introduction) /M.Okada 507 IL-28, IL-29 IFN-γ IL-10 IL-10 TGF-β IL-4 IL-25 IL-4 IL-5 IL-13 TNF-α 4 IL-23 IL-27 IL-31 IL-32 IL-7 IL-15 IL-17 IL-23 IL-27 IL-31 T DNA HSP65 DNA+ IL-12 DNA IL-32 Plos Path 2006 T 5 IFN-γ IFN-γ BCG M.avium IFN-γ IFN-γ 3) TNF-α TNF-α TNF TNF-Rp55 TNF / IL-6 IFN-γ IL-6 Mφ T IL-6 IL-6 / CRP IL-12 IL-12 IL-12 ril-12 BALB/c IL-12 IL-12p40 IL-12R β 1 IL-12 IL-23 IL-23R IL-12 IL-12R IL-23 / 6 IL-6 IL-6 DNA+IL-6 DNA+gp130 DNA T IL-2 IFN-γ DNA Condos 5 IFNγ 5 4 1 CT 1 1 3 IL-12 IFN-γ IL-2 IFN-α G-CSF 7 TNF-α TNF-α RA MCP-1 RANTES Ⅶ. おわりに Th17 regulatory T 文 1. 4,,, 2006, 50 _ 58. 2.. 2005 ; 60 : 678 _ 696. 3 state of arts, 献,, 2004, 209 _ 213. 4 Okada M, Kita Y, Nakajima T, et al.: Novel prophylactic and therapeutic vaccine against tuberculosis. Vaccine. 2009 ; 27 : 3267 _ 3270. 5 Yoshida S, Tanaka T, Kita Y, et al.: DNA vaccine using hemagglutinating virus of Japan-liposome encapsulating combination encoding mycobacterial heat shock protein 65 and interleukin-12 confers protection against Mycobacterium tuberculosis by T cell activation. Vaccine. 2006 ; 24 : 1191 _ 1204. 6 Okada M, Kita Y, Nakajima T, et al.: Evaluation of a novel vaccine (HVJ-liposome/HSP65 DNA+IL-12 DNA) against tuberculosis using the cynomolgus monkey model of TB. Vaccine. 2007 ; 25 : 2990 _ 2993. 7 Kita Y, Tanaka T, Yoshida S, et al.: Novel recombinant BCG and DNA-vaccination against tuberculosis in a cynomolgus monkey model. Vaccine. 2005 ; 23 : 2132 _ 2135. 8 Okada M, Kita Y : Tuberculosis vaccine development : The
508 85 6 2010 6 development of novel (preclinical) DNA vaccine. Human Vaccine. 2010 ; 6 : 1 _ 12. 9 Okada M, Kita Y, Nakajima T, et al.: A Novel Therapeutic and Prophylactic Vaccine (HVJ-Envelope/Hsp65 DNA+IL-12 DNA) against Tuberculosis Using the Cynomolgus Monkey Model. Procedia in Vaccinology. 2010 ; 2 : 34 _ 39. 10 Flynn JL, Chan J : Immunology of tuberculosis. Annu Rev Immunol. 2001 ; 19 : 93 _ 129. 11 Schluger NW, Rom WN : The host immune response to tuberculosis. Am J Respir Crit Care Med. 1998 ; 157 : 679 _ 691. 12 Tanaka F, Abe M, Akiyoshi T, et al.: The anti-human tumor effect and generation of human cytotoxic T cells in SCID mice given human peripheral blood lymphocytes by the in vivo transfer of the Interleukin-6 gene using adenovirus vector. Cancer Res. 1997 ; 57 : 1335 _ 1343. 13 Akira S : Toll-like receptors and innate immunity. Adv Immunol. 2001 ; 78 : 1 _ 56. 14, RNA MDA5 RIG-I. Annual Review.,, 2008, 57 _ 60. 15 Yanai H, Ban T, Wang Z, et al. : HMGB proteins function as universal sentinels for nucleic-acid-mediated innate immune responses. Nature. 2009 ; 462 : 99 _ 103. 16 Okada M, Yoshimura N, Kaieda T, et al. : Establishment and characterization of human T hybrid cells secreting immunoregulatory molecules. Proc Natl Acad Sci USA. 1981 ; 78 : 7718 _ 7721. 17 Okada M, Sakaguchi N, Yoshimura N, et al. : B cell growth factors and B cell differentiation factor from human T hybridomas. Two distinct kinds of B cell growth factor and their synergism in B cell proliferation. J Exp Med. 1983 ; 157 : 583 _ 590. 18 Okada M, Kitahara M, Kishimoto S, et al. : IL-6/BSF-2 functions as a killer helper factor in the in vitro induction of cytotoxic T cells. J Immunol. 1988 ; 141 : 1543 _ 1549. Current Topics IMMUNITY AGAINST MYCOBACTERIUM TUBERCULOSIS (INTRODUCTION) Masaji OKADA Abstract A third of world s population infected with Mycobacterium tuberculosis, and 2 million people die from tuberculosis every year. It is well established that protective to M. tuberculosis depends on both CD4 + and CD8 + T cells 1) 8). In particular, acquired immunity (cytotoxic T cell, Th1 helper T cell and Mφ) play an important role for TB infection. Recently, natural immunity also play a very attractive role for the development of TB immunity. We found that memory CTL is most important for the protection against TB using several kinds of mice. It was demonstrated that DBA/1 mice are more sensitive to TB infection than BALB/c mice (Th2 prone mice). Induction of memory CTL in DBA/1 mice was lower than BALB/c. In contrast, IFN-γ production of DBA/1 mice was higher than BALB/c. Therefore, famous researchers in the fields of TB immunity reviewed the recent advances of TB immunity, such as (1) T cell immunity and recognition against TB antigen, (2) TLR9 and CpG motif, (3) lipocalin2 and SLPI in natural TB immunity, (4) acquired immunity (CTL) and granulysin. The development novel vaccine (HSP65+IL-12 DNA vaccine), (5) The mechanism of protection against TB, in this mini-review series. Key words: Immunity against M. TB, T cell, Acguired immunity, Innate immunity Clinical Research Center, National Hospital Organization Kinki-chuo Chest Medical Center Correspondence to: Masaji Okada, Clinical Research Center, National Hospital Organization Kinki-chuo Chest Medical Center, 1180 Nagasone-cho, Kita-ku, Sakai-shi, Osaka 591 _ 8555 Japan. (E-mail: okm@kch.hosp.go.jp)