Limits...
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.

Show MeSH

Related in: MedlinePlus

Effect of C. caviae Vaccination on Histopathological Lesions in the Genital tract from Guinea pigs following Chlamydial Challenge.The genital tracts from each humanely euthanized guinea pig were removed at day 80 post C. caviae challenge fixed and embedded and then sectioned, and analyzed microscopically after H&E staining. (A) Representative photomicrographs of histological sections from uterine tissues are shown for each group of challenged guinea pigs with C. caviae EB vaccination (EB, n = 3) or mock vaccinated (Mock, n = 3). The superimposed images are magnifications of the regions of the indicated boxes to show details of inflammatory cell infiltration (c and f) and hemorrhage (a and d). Original magnification of the images (b and e) is ×200, while a, c, d and f are ×400. The light blue dash lines mark superficial layer exfoliation of the endometrial epithelium of the uterus (b), whereas the light blue solid lines indicate the intact endometrial epithelium of the uterus (e). Histopathological injury scores were calculated from five distinct morphological parameters (inflammatory cell infiltration, superficial layer exfoliation, edema, congestion and hemorrhage) in the uterus (B). Scores were calculated by examination of 5 consecutive sections (2 mm-interval) in every animal. Graphs expressed as mean ± SD, and compared using paired t- test. The asterisk indicates statistically significant differences (* p<0.05) between the C. caviae group and the mock group for the respective parameter.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4263467&req=5

pone-0114261-g004: Effect of C. caviae Vaccination on Histopathological Lesions in the Genital tract from Guinea pigs following Chlamydial Challenge.The genital tracts from each humanely euthanized guinea pig were removed at day 80 post C. caviae challenge fixed and embedded and then sectioned, and analyzed microscopically after H&E staining. (A) Representative photomicrographs of histological sections from uterine tissues are shown for each group of challenged guinea pigs with C. caviae EB vaccination (EB, n = 3) or mock vaccinated (Mock, n = 3). The superimposed images are magnifications of the regions of the indicated boxes to show details of inflammatory cell infiltration (c and f) and hemorrhage (a and d). Original magnification of the images (b and e) is ×200, while a, c, d and f are ×400. The light blue dash lines mark superficial layer exfoliation of the endometrial epithelium of the uterus (b), whereas the light blue solid lines indicate the intact endometrial epithelium of the uterus (e). Histopathological injury scores were calculated from five distinct morphological parameters (inflammatory cell infiltration, superficial layer exfoliation, edema, congestion and hemorrhage) in the uterus (B). Scores were calculated by examination of 5 consecutive sections (2 mm-interval) in every animal. Graphs expressed as mean ± SD, and compared using paired t- test. The asterisk indicates statistically significant differences (* p<0.05) between the C. caviae group and the mock group for the respective parameter.

Mentions: To evaluate the effect of C. caviae EB vaccination on development of pathological lesions in the genital tract, sections were obtained from challenged guinea pigs at day 80. Previous extensive analyses have demonstrated the suitability of this time-period to evaluate the upper genital tract sequelae following i.vag. Chlamydia challenge [12], [15], [43]. Histological analysis of the uterus 80 days after chlamydial challenge of mock-vaccinated animals revealed pathological damage that was characterized by the presence of a severe inflammatory cell infiltration (majority of the inflammatory cells were lymphocytes and macrophages) (Fig. 4A), moderate superficial layer exfoliation (Fig. 4A) and hemorrhage (Fig. 4A, a and b). In contrast, C. caviae EB vaccinated animals had an intact endometrial epithelium (Fig. 4A, e), reduced inflammation (Fig.4A, e and f) and hemorrhage in the uterus (Fig.4A, e and d). The mean histopathology severity scores for the uterus demonstrated significantly (p<0.05) reduced inflammatory cell infiltration, superficial layer exfoliation, and hemorrhage upon C. caviae EB vaccination compared to controls (Fig. 3B). Congestion and edema were reduced in vaccinated animals, but these scores were not statistically different from the mock-vaccinated guinea pigs.


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)

Effect of C. caviae Vaccination on Histopathological Lesions in the Genital tract from Guinea pigs following Chlamydial Challenge.The genital tracts from each humanely euthanized guinea pig were removed at day 80 post C. caviae challenge fixed and embedded and then sectioned, and analyzed microscopically after H&E staining. (A) Representative photomicrographs of histological sections from uterine tissues are shown for each group of challenged guinea pigs with C. caviae EB vaccination (EB, n = 3) or mock vaccinated (Mock, n = 3). The superimposed images are magnifications of the regions of the indicated boxes to show details of inflammatory cell infiltration (c and f) and hemorrhage (a and d). Original magnification of the images (b and e) is ×200, while a, c, d and f are ×400. The light blue dash lines mark superficial layer exfoliation of the endometrial epithelium of the uterus (b), whereas the light blue solid lines indicate the intact endometrial epithelium of the uterus (e). Histopathological injury scores were calculated from five distinct morphological parameters (inflammatory cell infiltration, superficial layer exfoliation, edema, congestion and hemorrhage) in the uterus (B). Scores were calculated by examination of 5 consecutive sections (2 mm-interval) in every animal. Graphs expressed as mean ± SD, and compared using paired t- test. The asterisk indicates statistically significant differences (* p<0.05) between the C. caviae group and the mock group for the respective parameter.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0114261-g004: Effect of C. caviae Vaccination on Histopathological Lesions in the Genital tract from Guinea pigs following Chlamydial Challenge.The genital tracts from each humanely euthanized guinea pig were removed at day 80 post C. caviae challenge fixed and embedded and then sectioned, and analyzed microscopically after H&E staining. (A) Representative photomicrographs of histological sections from uterine tissues are shown for each group of challenged guinea pigs with C. caviae EB vaccination (EB, n = 3) or mock vaccinated (Mock, n = 3). The superimposed images are magnifications of the regions of the indicated boxes to show details of inflammatory cell infiltration (c and f) and hemorrhage (a and d). Original magnification of the images (b and e) is ×200, while a, c, d and f are ×400. The light blue dash lines mark superficial layer exfoliation of the endometrial epithelium of the uterus (b), whereas the light blue solid lines indicate the intact endometrial epithelium of the uterus (e). Histopathological injury scores were calculated from five distinct morphological parameters (inflammatory cell infiltration, superficial layer exfoliation, edema, congestion and hemorrhage) in the uterus (B). Scores were calculated by examination of 5 consecutive sections (2 mm-interval) in every animal. Graphs expressed as mean ± SD, and compared using paired t- test. The asterisk indicates statistically significant differences (* p<0.05) between the C. caviae group and the mock group for the respective parameter.
Mentions: To evaluate the effect of C. caviae EB vaccination on development of pathological lesions in the genital tract, sections were obtained from challenged guinea pigs at day 80. Previous extensive analyses have demonstrated the suitability of this time-period to evaluate the upper genital tract sequelae following i.vag. Chlamydia challenge [12], [15], [43]. Histological analysis of the uterus 80 days after chlamydial challenge of mock-vaccinated animals revealed pathological damage that was characterized by the presence of a severe inflammatory cell infiltration (majority of the inflammatory cells were lymphocytes and macrophages) (Fig. 4A), moderate superficial layer exfoliation (Fig. 4A) and hemorrhage (Fig. 4A, a and b). In contrast, C. caviae EB vaccinated animals had an intact endometrial epithelium (Fig. 4A, e), reduced inflammation (Fig.4A, e and f) and hemorrhage in the uterus (Fig.4A, e and d). The mean histopathology severity scores for the uterus demonstrated significantly (p<0.05) reduced inflammatory cell infiltration, superficial layer exfoliation, and hemorrhage upon C. caviae EB vaccination compared to controls (Fig. 3B). Congestion and edema were reduced in vaccinated animals, but these scores were not statistically different from the mock-vaccinated guinea pigs.

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