The immunogenicity and protective effect of a DNA vaccine encoding the heat-shock protein (Hsp) GroEL of Chlamydophila abortus AB7, an obligate intracellular bacterium that causes abortion in sheep, was evaluated in pregnant and non-pregnant mouse models. The C. abortus groEL gene was cloned by screening a genomic library constructed in λFIX II arms with a nucleic acid probe corresponding to the central portion of the groEL gene from C. abortus. Sequence analysis of a positive clone revealed an open reading frame of 1632 bp encoding a 544 amino acid polypeptide with a predicted molecular mass of 58 256 Da and highly similar to GroEL of Chlamydia trachomatis (93 %) and Chlamydophila pneumoniae (94 %). As observed in other sequenced chlamydial genomes, the groEL gene belongs to an operon comprising another gene encoding the Hsp GroES. OF1 outbred mice were immunized intramuscularly with plasmid DNA carrying the groEL gene three times at 3 week intervals and challenged 2 weeks after the last DNA injection. In pregnant mice, no reduction in abortion was observed and the DNA vaccination failed to reduce the bacterial infection in the placenta and spleen of mice. Nevertheless, partial protection of fetuses was obtained. Immunization of non-pregnant mice with the groEL gene resulted in a specific humoral response with the predominant IgG2a isotype, suggesting a Th1-type immune response. The anti-GroEL antibodies showed no neutralizing effect in vitro on C. abortus infectivity. Although the DNA vaccine induced a delayed-type hypersensitivity response, it failed to elicit an efficient cellular immune response since the mice were not protected against bacterial challenge.
BoumedineK. S.,
RodolakisA.1998; AFLP allows the identification of genomic markers of ruminant Chlamydia psittaci strains useful for typing and epidemiological studies. Res Microbiol 149:735–744[CrossRef]
BuendíaA. J.,
SánchezJ.,
Del RioL.,
GarcésB.,
GallegoM. C.,
CaroM. R.,
BernabéA.,
SalinasJ.1999; Differences in the immune response against ruminant chlamydial strains in a murine model. Vet Res 30:495–507
Buzoni-GatelD.,
RodolakisA.,
PlommetM.1987; T cell mediated and humoral immunity in a mouse Chlamydia psittaci systemic infection. Res Vet Sci 43:59–63
CaldwellH. D.,
KromhoutJ.,
SchachterJ.1981; Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis
. Infect Immun 31:1161–1176
DavisH. L.,
WhalenR. G.,
DemeneixB. A.1993; Direct gene transfer into skeletal muscle in vivo : factors affecting efficiency of transfer and stability of expression. Hum Gene Ther 4:151–159[CrossRef]
Dong-JiZ.,
YangX.,
ShenC.,
LuH.,
MurdinA.,
BrunhamR. C.2000; Priming with Chlamydia trachomatis major outer membrane protein (MOMP) DNA followed by MOMP ISCOM boosting enhances protection and is associated with increased immunoglobulin A and Th1 cellular immune responses. Infect Immun 68:3074–3078[CrossRef]
FayeP.,
ChartonA.,
MageC.,
BernardC.,
Le LayecC.1972; Propriétés hémagglutinantes du ‘‘virus’’ de l'avortement enzootique des petits ruminants (souches de ‘‘Rakeia’’ d'origine ovine et caprine).Note préliminaire. Bull Acad Vét Fr 45:169–173 (in French
HasanU. A.,
AbaiA. M.,
HarperD. R.,
WrenB. W.,
MorrowW. J.1999; Nucleic acid immunization: concepts and techniques associated with third generation vaccines. J Immunol Methods 229:1–22[CrossRef]
HéchardC.,
GrépinetO.,
RodolakisA.2002; Protection evaluation against Chlamydophila abortus challenge by DNA vaccination with a dnaK -encoding plasmid in pregnant and non-pregnant mice. Vet Res 33:313–326[CrossRef]
HéchardC.,
GrépinetO.,
RodolakisA.2003; Evaluation of protection against Chlamydophila abortus challenge after DNA immunization with the major outer-membrane protein-encoding gene in pregnant and non-pregnant mice. J Med Microbiol 52:35–40[CrossRef]
LeclerqS.,
HarmsJ. S.,
RosinhaG. M.,
AzevedoV.,
OliveiraS. C.2002; Induction of a Th1-type of immune response but not protective immunity by intramuscular DNA immunisation with Brucella abortus GroEL heat-shock gene. J Med Microbiol 51:20–26
MarckC.1988; ‘‘DNA Strider’': a ‘‘C’’ program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers. Nucleic Acids Res 16:1829–1836[CrossRef]
MurdinA. D.,
SuH.,
KleinM. H.,
CaldwellH. D.1995; Poliovirus hybrids expressing neutralization epitopes from variable domains I and IV of the major outer membrane protein of Chlamydia trachomatis elicit broadly cross-reactive C.trachomatis -neutralizing antibodies. Infect Immun 63:1116–1121
PappJ. R.,
ShewenP. E.1996; Localization of chronic Chlamydia psittaci infection in the reproductive tract of sheep. J Infect Dis 174:1296–1302[CrossRef]
PenttiläT.,
VuolaJ. M.,
PuurulaV.,
AnttilaM.,
SarvasM.,
RautonenN.,
MäkeläP. H.,
PuolakkainenM.2000; Immunity to Chlamydia pneumoniae induced by vaccination with DNA vectors expressing a cytoplasmic protein (Hsp60) or outer membrane proteins (MOMP and Omp2). Vaccine 19:1256–1265[CrossRef]
RodolakisA.1983; In vitro and in vivo properties of chemically induced temperature-sensitive mutants of Chlamydia psittac i var. ovis : screening in a murine model. Infect Immun 42:525–530
RodolakisA.,
SouriauA.1983; Response of ewes to temperature-sensitive mutants of Chlamydia psittaci (var ovis ) obtained by NTG mutagenesis. Ann Rech Vet 14:155–161
RodolakisA.,
SouriauA.1986; Response of goats to vaccination with temperature-sensitive mutants of Chlamydia psittaci obtained by nitrosoguanidine mutagenesis. Am J Vet Res 47:2627–2631
RodolakisA.,
GestinL.,
BertinA.1981; Method of control of vaccines against ovine abortive chlamydiosis using pregnant mice. Ann Rech Vet 12:371–377 (in French
SrivastavaP. K.,
MenoretA.,
BasuS.,
BinderR. J.,
McQuadeK. L.1998; Heat shock proteins come of age: primitive functions acquire new roles in an adaptive world. Immunity 8:657–665[CrossRef]
TasconR. E.,
ColstonM. J.,
RagnoS.,
StavropoulosE.,
GregoryD.,
LowrieD. B.1996; Vaccination against tuberculosis by DNA injection. Nat Med 2:888–892[CrossRef]
WideraG.,
AustinM.,
RabussayD. & 8 other authors; 2000; Increased DNA vaccine delivery and immunogenicity by electroporation in vivo
. J Immunol 164:4635–4640[CrossRef]
WolffJ. A.,
MaloneR. W.,
WilliamsP.,
ChongW.,
AcsadiG.,
JaniA.,
FelgnerP. L.1990; Direct gene transfer into mouse muscle in vivo
. Science 247:1465–1468[CrossRef]
YuanY.,
LyngK.,
ZhangY. X.,
RockeyD. D.,
MorrisonR. P.1992; Monoclonal antibodies define genus-specific, species-specific, and cross-reactive epitopes of the chlamydial 60-kilodalton heat shock protein (hsp60): specific immunodetection and purification of chlamydial hsp60. Infect Immun 60:2288–2296