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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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C. caviae Immune Serum and Chlamydial Infectivity.Sera obtained from guinea pigs (n = 5 per group) 30 days post vaccination with C. caviae or mock (PBS) or 15 days after challenge (+ challenge) were heat-inactivated (HK) at 56°C for 30 min or left untreated and diluted 1∶25 in DMEM before addition into 96 well plate. Sera from each animal was incubated with C. caviae (2×104 IFU) for 1 hr in a shaking incubator, then sera/bacterial mixtures were added to HeLa cells (0.01 MOI) for bacterial enumeration. Bacterial numbers for each group were presented as a box-and-whisker plot. * p<0.05.
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pone-0114261-g003: C. caviae Immune Serum and Chlamydial Infectivity.Sera obtained from guinea pigs (n = 5 per group) 30 days post vaccination with C. caviae or mock (PBS) or 15 days after challenge (+ challenge) were heat-inactivated (HK) at 56°C for 30 min or left untreated and diluted 1∶25 in DMEM before addition into 96 well plate. Sera from each animal was incubated with C. caviae (2×104 IFU) for 1 hr in a shaking incubator, then sera/bacterial mixtures were added to HeLa cells (0.01 MOI) for bacterial enumeration. Bacterial numbers for each group were presented as a box-and-whisker plot. * p<0.05.

Mentions: Vaccination with C. caviae EB produced elevated levels of total Chlamydia-binding IgG, compared to mock vaccinated guinea pigs at day 30 post-vaccination (Fig. 2A). However, anti-C. caviae serum titers were comparable in C. caviae and mock vaccinated guinea pigs at 15 days post challenge (Fig. 2B). Given that a significant difference in bacterial shedding between vaccinated and mock vaccinated animals was observed (Fig. 1), we next compared the neutralization capacity of sera collected from both groups of guinea pigs. As shown in Fig. 3, sera collected day 30 post vaccination from C. caviae EB vaccinated guinea pigs reduced infectivity in HeLa cells by approximately two logs compared to sera from mock vaccinated guinea pigs. This difference in ability to neutralize C. caviae in vitro increased to approximately three logs at day 15 post-challenge, supporting the conclusion that sera from C. caviae EB vaccinated guinea pigs generated antibodies capable of neutralizing C. caviae better than mock immunized animals. Because similar trends were observed in heat-treated sera we conclude that the neutralizing effects observed were independent of complement.


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)

C. caviae Immune Serum and Chlamydial Infectivity.Sera obtained from guinea pigs (n = 5 per group) 30 days post vaccination with C. caviae or mock (PBS) or 15 days after challenge (+ challenge) were heat-inactivated (HK) at 56°C for 30 min or left untreated and diluted 1∶25 in DMEM before addition into 96 well plate. Sera from each animal was incubated with C. caviae (2×104 IFU) for 1 hr in a shaking incubator, then sera/bacterial mixtures were added to HeLa cells (0.01 MOI) for bacterial enumeration. Bacterial numbers for each group were presented as a box-and-whisker plot. * p<0.05.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0114261-g003: C. caviae Immune Serum and Chlamydial Infectivity.Sera obtained from guinea pigs (n = 5 per group) 30 days post vaccination with C. caviae or mock (PBS) or 15 days after challenge (+ challenge) were heat-inactivated (HK) at 56°C for 30 min or left untreated and diluted 1∶25 in DMEM before addition into 96 well plate. Sera from each animal was incubated with C. caviae (2×104 IFU) for 1 hr in a shaking incubator, then sera/bacterial mixtures were added to HeLa cells (0.01 MOI) for bacterial enumeration. Bacterial numbers for each group were presented as a box-and-whisker plot. * p<0.05.
Mentions: Vaccination with C. caviae EB produced elevated levels of total Chlamydia-binding IgG, compared to mock vaccinated guinea pigs at day 30 post-vaccination (Fig. 2A). However, anti-C. caviae serum titers were comparable in C. caviae and mock vaccinated guinea pigs at 15 days post challenge (Fig. 2B). Given that a significant difference in bacterial shedding between vaccinated and mock vaccinated animals was observed (Fig. 1), we next compared the neutralization capacity of sera collected from both groups of guinea pigs. As shown in Fig. 3, sera collected day 30 post vaccination from C. caviae EB vaccinated guinea pigs reduced infectivity in HeLa cells by approximately two logs compared to sera from mock vaccinated guinea pigs. This difference in ability to neutralize C. caviae in vitro increased to approximately three logs at day 15 post-challenge, supporting the conclusion that sera from C. caviae EB vaccinated guinea pigs generated antibodies capable of neutralizing C. caviae better than mock immunized animals. Because similar trends were observed in heat-treated sera we conclude that the neutralizing effects observed were independent of complement.

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