The groES gene of Mycobacterium avium strain 485 was cloned and expressed in Escherichia coli and the recombinant GroES protein was purified by affinity chromatography. The GroES preparation showed high purity by electrophoresis and immunoblotting. Immuno-electron microscopy showed that GroES was located both in the cytoplasm and on the surface of the mycobacterial cells and thus is readily available to interact with the host immune system. BALB/c mice were immunised intranasally with recombinant GroES, alone or in combination with a synthetic oligodeoxynucleotide containing unmethylated CpG motifs, and tested for protection against infection with M. avium. Neither GroES nor CpG alone provided any protection against subsequent challenge with M. avium, whereas a combination of the two significantly protected the lungs and spleen against colonisation by M. avium after intranasal challenge with a low dose of the organism. This indicates that intranasal administration of GroES and CpG oligodeoxynucleotides increases the resistance of BALB/c mice to M. avium infection.
HorshburgCR. Epidemiology of Mycobacterium avium complex. In
KorvickJA,
BensonCA.
edsMycobacterium avium complex infection: progress in research and treatmentM. Dekker19961–22
MiddletonAM,
ChadwickMV,
NicholsonAG. et al. The role of Mycobacterium avium complex fibronectin attachment protein in adherence to the human respiratory mucosa. Mol Microbiol2000; 38:381–391[CrossRef]
von ReynCF,
WaddelRD,
EatonT. et al. Isolation of Mycobacterium avium complex from water in the United States, Finland, Zaire and Kenya. J Clin Microbiol1993; 31:3227–3230
VerbonA,
KuijperS,
JansenHM,
SpeelmanP,
KolkAH. Antigens in culture supernatant of Mycobacterium tuberculosis : epitopes defined by monoclonal and human antibodies. J Gen Microbiol1990; 136:955–964[CrossRef]
OrmeIM,
MillerES,
RobertsAD. et al. T lymphocytes mediating protection and cellular cytolysis during the course of Mycobacterium tuberculosis infection.Evidence for different kinetics and recognition of a wide spectrum of protein antigens. J Immunol1992; 148:189–196
GelberRH,
MehraV,
BloomB. et al. Vaccination with pure Mycobacterium leprae proteins inhibits M. leprae multiplication in mouse footpads. Infect Immun1994; 10:4250–4255
McCluskieMJ,
DavisHL. CpG DNA is a potent enhancer of systemic and mucosal immune responses against hepatitis B surface antigen with intranasal administration to mice. J Immunol1998; 161:4463–4466
FattoriniL,
XiaoY,
AusielloCM. et al. Late acquisition of hyporesponsiveness to lipopolysaccharide by Mycobacterium avium -infected human macrophages in producing tumor necrosis factor-alpha but not interleukin-1 beta and -6. J Infect Dis1996; 173:1030–1034[CrossRef]
MolinariA,
CalcabriniA,
MeschiniS. et al. Detection of P-glycoprotein in the Golgi apparatus of drug-untreated human melanoma cells. Int J Cancer1998; 75:885–893[CrossRef]
von HunolsteinC,
MariottiS,
TeloniR. et al. The adjuvant effect of synthetic oligodeoxynucleotide containing CpG motif converts the anti- Haemophilus influenzae type b glycoconjugates into efficient anti-polysaccharide and anti-carrier polyvalent vaccines. Vaccine2001; 19:3058–3066[CrossRef]
Rinke de WitTF, BekelieS,
OslandA. et al. Mycobacteria contain two groEL genes: the second Mycobacterium leprae groEL gene is arranged in an operon with groES . Mol Microbiol1992; 6:1995–2007[CrossRef]
ZuberU,
SchumannW. CIRCE, a novel heat shock element involved in regulation of heat shock operon dnaK of Bacillus subtilis . J Bacteriol1994; 176:1359–1363
SchonU,
SchumannW. Construction of His6-tagging vectors allowing single-step purification of GroES and other polypeptides produced in Bacillus subtilis . Gene1994; 147:91–94[CrossRef]
MoldoveanuZ,
Love-HomanL,
HuangWQ,
KriegAM. CpG DNA, a novel immune enhancer for systemic and mucosal immunization with influenza virus. Vaccine1998; 16:1216–1224[CrossRef]
FreidagBL,
MeltonGB,
CollinsF. et al. CpG oligodeoxynucleotides and interleukin-12 improve the efficacy of Mycobacterium bovis BCG vaccination in mice challenged with M. tuberculosis . Infect Immun2000; 68:2948–2953[CrossRef]