Development of therapeutic and prophylactic vaccine against Tuberculosis using monkey and transgenic mice models.
OBJECTIVE
BCG is not efficacious against M. tuberculosis (TB) in adult. Therefore, novel TB vaccines were established by using three kinds of animal models (cynomolgus monkey model which is the best animal model of human TB, IL-2R knock out SCID mice as a human immune model, and granulysin transgenic mouse).


RESULTS
DNA vaccine expressing TB Hsp65 and IL-12 was delivered by the hemagglutinating virus of Japan (HVJ)-envelope. The BCG prime followed by Hsp65+IL-12/HVJ vaccine boost showed a synergistic effect in the TB-infected cynomolgus monkey (100% survival). In contrast, 33% of monkeys were alive in BCG alone group. Furthermore, the prolongation of survival period of the monkey was observed by the combination of BCG and DNA vaccine even when the boost was performed after long-term period (4month) from prime. This combination also improved the erythrocyte sedimentation rate (ESR), increased the body weight, and augmented the proliferation of PBL and IL-12 production at higher levels than BCG alone or saline. Furthermore, this vaccine exerted therapeutic efficacy in IL-2R knock out SCID-PBL/hu mice, which were transplanted with human T cells. Granulysin is an important defensive molecule expressed by human T cells and NK cells and has a cytolytic activity against microbes including Mycobacterium tuberculosis (TB) and tumors. Expression of 15kD (15K) granulysin protein and mRNA in CD8 positive T cells in the patients infected with drug sensitive (TB) or multi-drug resistant (MDR-TB) M. tuberculosis were lower than that in the healthy volunteers, suggesting that granulysin treatment might improve the tuberculous disease in human. Therefore, we established two kinds of granulysin transgenic mice (15K granulysin transgenic mice and 9K granulysin transgenic mice). It was demonstrated that 15K granulysin transgenic mice as well as 9K granulysin transgenic mice exerted in vivo anti-TB effect, including the decrease of the number of TB and augmentation of the CTL activity. These are the first findings which demonstrate in vivo effects of 15K granulysin and 9K granulysin against TB infection. Moreover, DNA vaccine expressing 15K granulysin showed a therapeutic activity against TB in mice.


CONCLUSIONS
These data indicate that monkey, IL-2R gene-knock out SCID-PBL/hu and granulysin transgenic mice models provide useful tools for the development of novel vaccines (HVJ-Envelope/Hsp65 DNA + IL-12 DNA vaccine and granulysin vaccine) against TB.
Clinical Research Center, National Hospital Organization Kinki-chuo Chest Medical Center, Kitaku, Sakai, Osaka, Japan.
Yoko
Y
Kita
Masaji
M
Okada
Toshihiro
T
Nakajima
Noriko
N
Kanamaru
Satomi
S
Hashimoto
Tetsuji
T
Nagasawa
Yasufumi
Y
Kaneda
Shigeto
S
Yoshida
Yasuko
Y
Nishida
Hitoshi
H
Nakatani
Kyoko
K
Takao
Chie
C
Kishigami
Shiho
S
Nishimatsu
Yuki
Y
Sekine
Yasushi
Y
Takamori
David N
DN
McMurray
E C
EC
De la Cruz
E V
EV
Tan
R M
RM
Abalos
J A
JA
Burgos
Paul
P
Saunderson
Mitsunori
M
Sakatani
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D049109:Cell Proliferation; D018834:Chaperonin 60 / Q000235:genetics / Q000276:immunology*; D004195:Disease Models, Animal; D007117:Immunization, Secondary / Q000379:methods; D018664:Interleukin-12 / Q000235:genetics / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008252:Macaca fascicularis; D051379:Mice; D016513:Mice, SCID; D008822:Mice, Transgenic; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology*; D018419:Primate Diseases / Q000276:immunology / Q000517:prevention & control; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012376:Rodent Diseases / Q000276:immunology / Q000517:prevention & control; D032581:Tuberculosis Vaccines / Q000008:administration & dosage* / Q000276:immunology*; D014611:Vaccination / Q000379:methods
D001426:Bacterial Proteins; D018834:Chaperonin 60; D011994:Recombinant Proteins; D032581:Tuberculosis Vaccines; C074119:heat-shock protein 65, Mycobacterium; D018664:Interleukin-12