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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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Comparative Heatmap Depiction of Differential Gene Expression using RT-PCR array screening.Three groups of guinea pigs were used for the comparative study: non vaccinated and non challenged (naïve), mock vaccinated but challenged (PBS/C), and C. caviae EB vaccinated and challenged (EB/C) groups. Each group contained three animals. The tissues (upper and lower genital tracts, U and L, respectively) from the respective groups of animals were collected at days 3 and 9 after challenge. Red shading indicates an increase in expression, while blue shading indicates suppression of expression, of the gene indicated on the right side of the panel. Lighter shades including white indicate similar levels of expression. Functional gene clustering is indicated by the brackets on the left showing 3 major groups consisting of innate, Th2 and Th1/inflammatory related genes.
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pone-0114261-g005: Comparative Heatmap Depiction of Differential Gene Expression using RT-PCR array screening.Three groups of guinea pigs were used for the comparative study: non vaccinated and non challenged (naïve), mock vaccinated but challenged (PBS/C), and C. caviae EB vaccinated and challenged (EB/C) groups. Each group contained three animals. The tissues (upper and lower genital tracts, U and L, respectively) from the respective groups of animals were collected at days 3 and 9 after challenge. Red shading indicates an increase in expression, while blue shading indicates suppression of expression, of the gene indicated on the right side of the panel. Lighter shades including white indicate similar levels of expression. Functional gene clustering is indicated by the brackets on the left showing 3 major groups consisting of innate, Th2 and Th1/inflammatory related genes.

Mentions: We developed a guinea pig-specific array (S2 Figure), to screen for differences in selected immune response-related genes involved in C. caviae EB vaccination-induced protection (Fig. 1). Results of the qRT-PCR analyses revealed modulation of 19 highly regulated genes that were subjected to hierarchical clustering analyses for probable co-regulation of immune components in C. caviae EB vaccinated or mock vaccinated guinea pigs (Fig. 5). The co-regulation of innate (NK), Th2-humoral (including CD93, CD39, IL-4R, β2-microglobulin) and Th1-cellular/inflammatory responses was evident in the heat map as clustering into 3 major groups. Overall, following C. caviae infection (Fig. 5, lanes 1–4) NK activation genes (CD94, IL-21, and CD233) were upregulated. In contrast, Th2 humoral response-related genes were down-regulated and correlated to an increased Th1 cellular and inflammatory response by day 9.


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)

Comparative Heatmap Depiction of Differential Gene Expression using RT-PCR array screening.Three groups of guinea pigs were used for the comparative study: non vaccinated and non challenged (naïve), mock vaccinated but challenged (PBS/C), and C. caviae EB vaccinated and challenged (EB/C) groups. Each group contained three animals. The tissues (upper and lower genital tracts, U and L, respectively) from the respective groups of animals were collected at days 3 and 9 after challenge. Red shading indicates an increase in expression, while blue shading indicates suppression of expression, of the gene indicated on the right side of the panel. Lighter shades including white indicate similar levels of expression. Functional gene clustering is indicated by the brackets on the left showing 3 major groups consisting of innate, Th2 and Th1/inflammatory related genes.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0114261-g005: Comparative Heatmap Depiction of Differential Gene Expression using RT-PCR array screening.Three groups of guinea pigs were used for the comparative study: non vaccinated and non challenged (naïve), mock vaccinated but challenged (PBS/C), and C. caviae EB vaccinated and challenged (EB/C) groups. Each group contained three animals. The tissues (upper and lower genital tracts, U and L, respectively) from the respective groups of animals were collected at days 3 and 9 after challenge. Red shading indicates an increase in expression, while blue shading indicates suppression of expression, of the gene indicated on the right side of the panel. Lighter shades including white indicate similar levels of expression. Functional gene clustering is indicated by the brackets on the left showing 3 major groups consisting of innate, Th2 and Th1/inflammatory related genes.
Mentions: We developed a guinea pig-specific array (S2 Figure), to screen for differences in selected immune response-related genes involved in C. caviae EB vaccination-induced protection (Fig. 1). Results of the qRT-PCR analyses revealed modulation of 19 highly regulated genes that were subjected to hierarchical clustering analyses for probable co-regulation of immune components in C. caviae EB vaccinated or mock vaccinated guinea pigs (Fig. 5). The co-regulation of innate (NK), Th2-humoral (including CD93, CD39, IL-4R, β2-microglobulin) and Th1-cellular/inflammatory responses was evident in the heat map as clustering into 3 major groups. Overall, following C. caviae infection (Fig. 5, lanes 1–4) NK activation genes (CD94, IL-21, and CD233) were upregulated. In contrast, Th2 humoral response-related genes were down-regulated and correlated to an increased Th1 cellular and inflammatory response by day 9.

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