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Use of a Guinea pig-specific transcriptome array for evaluation of protective immunity against genital chlamydial infection following intranasal vaccination in Guinea pigs.

Wali S, Gupta R, Veselenak RL, Li Y, Yu JJ, Murthy AK, Cap AP, Guentzel MN, Chambers JP, Zhong G, Rank RG, Pyles RB, Arulanandam BP - PLoS ONE (2014)

Bottom Line: Importantly, at day 80 post challenge, vaccinated guinea pigs experienced significantly reduced genital pathology - a sequelae of genital chlamydial infections, in comparison to mock vaccinated guinea pigs.Th1-cellular/inflammatory immune genes and Th2-humoral associated genes were also found to be elevated in vaccinated guinea pigs at day 3 post-challenge and correlated with early clearance of the bacterium.Overall, this study provides the first evidence of guinea pig-specific genes involved in anti-chlamydial vaccination and illustrates the enhancement of the utility of this animal model in chlamydial pathogenesis.

View Article: PubMed Central - PubMed

Affiliation: South Texas Center for Emerging Infectious Diseases and Center of Excellence in Infection Genomics, University of Texas at San Antonio, One UTSA Circle, San Antonio, Texas 78249, United Stats of America.

ABSTRACT
Guinea pigs have been used as a second animal model to validate putative anti-chlamydial vaccine candidates tested in mice. However, the lack of guinea pig-specific reagents has limited the utility of this animal model in Chlamydia sp. vaccine studies. Using a novel guinea pig-specific transcriptome array, we determined correlates of protection in guinea pigs vaccinated with Chlamydia caviae (C. caviae) via the intranasal route, previously reported by us and others to provide robust antigen specific immunity against subsequent intravaginal challenge. C. caviae vaccinated guinea pigs resolved genital infection by day 3 post challenge. In contrast, mock vaccinated animals continued to shed viable Chlamydia up to day 18 post challenge. Importantly, at day 80 post challenge, vaccinated guinea pigs experienced significantly reduced genital pathology - a sequelae of genital chlamydial infections, in comparison to mock vaccinated guinea pigs. Sera from vaccinated guinea pigs displayed antigen specific IgG responses and increased IgG1 and IgG2 titers capable of neutralizing GPIC in vitro. Th1-cellular/inflammatory immune genes and Th2-humoral associated genes were also found to be elevated in vaccinated guinea pigs at day 3 post-challenge and correlated with early clearance of the bacterium. Overall, this study provides the first evidence of guinea pig-specific genes involved in anti-chlamydial vaccination and illustrates the enhancement of the utility of this animal model in chlamydial pathogenesis.

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Quantitative PCR Assessment of Bacterial Genomic Burdens in Lower and Upper Genital Tracts from C. caviae Mock or (PBS) Vaccinated Guinea Pigs.Groups of three animals were euthanized on days 3 and 9 after C. caviae i.vag. challenge and tissues representing the lower or upper genital tract were aseptically collected. DNA was subjected to qPCR for GAPDH (host target used for normalization) and the single copy C. caviae tryptophan synthase gene (quantification of bacterial load). The average bacterial burdens for each tissue are depicted as grey (mock-vaccinated) or black (vaccinated) bars for each tissue and time point. LGT: lower genital tract. UGT: upper genital tract. * p<0.05, ** p<0.01.
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pone-0114261-g006: Quantitative PCR Assessment of Bacterial Genomic Burdens in Lower and Upper Genital Tracts from C. caviae Mock or (PBS) Vaccinated Guinea Pigs.Groups of three animals were euthanized on days 3 and 9 after C. caviae i.vag. challenge and tissues representing the lower or upper genital tract were aseptically collected. DNA was subjected to qPCR for GAPDH (host target used for normalization) and the single copy C. caviae tryptophan synthase gene (quantification of bacterial load). The average bacterial burdens for each tissue are depicted as grey (mock-vaccinated) or black (vaccinated) bars for each tissue and time point. LGT: lower genital tract. UGT: upper genital tract. * p<0.05, ** p<0.01.

Mentions: In order to add to the bacterial shedding profiles of C. caviae EB and mock vaccinated animals (Fig. 1), bacterial burdens in lower and upper genital tract of guinea pigs at days 3 and 9 post challenge were estimated using qRT-PCR. Although C. caviae loads were comparable at day 3 post challenge in the lower and upper tract of vaccinated and mock-vaccinated guinea pigs (Fig. 6), by day 9, C. caviae EB vaccinated guinea pigs displayed 4–6 logs fewer bacterial genomes in lower and upper genital tracts. Additionally, of the twelve tissues from C. caviae EB vaccinated animals, three had no detectable bacterial genomes. Taken together, bacterial burdens (Fig. 6) and shedding data (Fig. 1) indicated a robust protection and generation of immune responses (Fig. 5, Table 1 and 2), were induced in C. caviae EB vaccinated guinea pigs compared to mock vaccination.


Use of a Guinea pig-specific transcriptome array for evaluation of protective immunity against genital chlamydial infection following intranasal vaccination in Guinea pigs.

Wali S, Gupta R, Veselenak RL, Li Y, Yu JJ, Murthy AK, Cap AP, Guentzel MN, Chambers JP, Zhong G, Rank RG, Pyles RB, Arulanandam BP - PLoS ONE (2014)

Quantitative PCR Assessment of Bacterial Genomic Burdens in Lower and Upper Genital Tracts from C. caviae Mock or (PBS) Vaccinated Guinea Pigs.Groups of three animals were euthanized on days 3 and 9 after C. caviae i.vag. challenge and tissues representing the lower or upper genital tract were aseptically collected. DNA was subjected to qPCR for GAPDH (host target used for normalization) and the single copy C. caviae tryptophan synthase gene (quantification of bacterial load). The average bacterial burdens for each tissue are depicted as grey (mock-vaccinated) or black (vaccinated) bars for each tissue and time point. LGT: lower genital tract. UGT: upper genital tract. * p<0.05, ** p<0.01.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263467&req=5

pone-0114261-g006: Quantitative PCR Assessment of Bacterial Genomic Burdens in Lower and Upper Genital Tracts from C. caviae Mock or (PBS) Vaccinated Guinea Pigs.Groups of three animals were euthanized on days 3 and 9 after C. caviae i.vag. challenge and tissues representing the lower or upper genital tract were aseptically collected. DNA was subjected to qPCR for GAPDH (host target used for normalization) and the single copy C. caviae tryptophan synthase gene (quantification of bacterial load). The average bacterial burdens for each tissue are depicted as grey (mock-vaccinated) or black (vaccinated) bars for each tissue and time point. LGT: lower genital tract. UGT: upper genital tract. * p<0.05, ** p<0.01.
Mentions: In order to add to the bacterial shedding profiles of C. caviae EB and mock vaccinated animals (Fig. 1), bacterial burdens in lower and upper genital tract of guinea pigs at days 3 and 9 post challenge were estimated using qRT-PCR. Although C. caviae loads were comparable at day 3 post challenge in the lower and upper tract of vaccinated and mock-vaccinated guinea pigs (Fig. 6), by day 9, C. caviae EB vaccinated guinea pigs displayed 4–6 logs fewer bacterial genomes in lower and upper genital tracts. Additionally, of the twelve tissues from C. caviae EB vaccinated animals, three had no detectable bacterial genomes. Taken together, bacterial burdens (Fig. 6) and shedding data (Fig. 1) indicated a robust protection and generation of immune responses (Fig. 5, Table 1 and 2), were induced in C. caviae EB vaccinated guinea pigs compared to mock vaccination.

Bottom Line: Importantly, at day 80 post challenge, vaccinated guinea pigs experienced significantly reduced genital pathology - a sequelae of genital chlamydial infections, in comparison to mock vaccinated guinea pigs.Th1-cellular/inflammatory immune genes and Th2-humoral associated genes were also found to be elevated in vaccinated guinea pigs at day 3 post-challenge and correlated with early clearance of the bacterium.Overall, this study provides the first evidence of guinea pig-specific genes involved in anti-chlamydial vaccination and illustrates the enhancement of the utility of this animal model in chlamydial pathogenesis.

View Article: PubMed Central - PubMed

Affiliation: South Texas Center for Emerging Infectious Diseases and Center of Excellence in Infection Genomics, University of Texas at San Antonio, One UTSA Circle, San Antonio, Texas 78249, United Stats of America.

ABSTRACT
Guinea pigs have been used as a second animal model to validate putative anti-chlamydial vaccine candidates tested in mice. However, the lack of guinea pig-specific reagents has limited the utility of this animal model in Chlamydia sp. vaccine studies. Using a novel guinea pig-specific transcriptome array, we determined correlates of protection in guinea pigs vaccinated with Chlamydia caviae (C. caviae) via the intranasal route, previously reported by us and others to provide robust antigen specific immunity against subsequent intravaginal challenge. C. caviae vaccinated guinea pigs resolved genital infection by day 3 post challenge. In contrast, mock vaccinated animals continued to shed viable Chlamydia up to day 18 post challenge. Importantly, at day 80 post challenge, vaccinated guinea pigs experienced significantly reduced genital pathology - a sequelae of genital chlamydial infections, in comparison to mock vaccinated guinea pigs. Sera from vaccinated guinea pigs displayed antigen specific IgG responses and increased IgG1 and IgG2 titers capable of neutralizing GPIC in vitro. Th1-cellular/inflammatory immune genes and Th2-humoral associated genes were also found to be elevated in vaccinated guinea pigs at day 3 post-challenge and correlated with early clearance of the bacterium. Overall, this study provides the first evidence of guinea pig-specific genes involved in anti-chlamydial vaccination and illustrates the enhancement of the utility of this animal model in chlamydial pathogenesis.

Show MeSH
Related in: MedlinePlus