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Immunogenicity without Efficacy of an Adenoviral Tuberculosis Vaccine in a Stringent Mouse Model for Immunotherapy during Treatment.

Alyahya SA, Nolan ST, Smith CM, Bishai WR, Sadoff J, Lamichhane G - PLoS ONE (2015)

Bottom Line: Even though immunotherapy resulted in strong splenic IFN-γ responses, no effect on bacterial replication in the lungs was seen.Multiplex assay analysis of lung samples revealed the absence of cytokine augmentation such as IFN-γ, TNF-α and IL-2, suggesting that immunization failed to induce immunity in the lungs.The results obtained from our study raise questions regarding the traits of protective TB immunity that are relevant for the development of future immunotherapeutic and post-exposure vaccination strategies.

View Article: PubMed Central - PubMed

Affiliation: Crucell Holland B.V., Janssen Infectious Diseases and Vaccines, Leiden, The Netherlands.

ABSTRACT
To investigate if bacterial persistence during TB drug treatment could be overcome by modulation of host immunity, we adapted a clinically-relevant model developed for the evaluation of new drugs and examined if immunotherapy with two adenoviral vaccines, Ad35-TBS (AERAS-402) and Ad26-TBS, could shorten therapy in mice. Even though immunotherapy resulted in strong splenic IFN-γ responses, no effect on bacterial replication in the lungs was seen. Multiplex assay analysis of lung samples revealed the absence of cytokine augmentation such as IFN-γ, TNF-α and IL-2, suggesting that immunization failed to induce immunity in the lungs. In this model, we show that IFN-γ levels were not associated with protection against disease relapse. The results obtained from our study raise questions regarding the traits of protective TB immunity that are relevant for the development of future immunotherapeutic and post-exposure vaccination strategies.

No MeSH data available.


Related in: MedlinePlus

Probing the mechanism of failure at week 28.A) ELISpot analysis was conducted at the final relapse time point of week 28 as previously described. Results of groups that received immunotherapy (TV Ad35+ and TV Ad26+) and the main negative control group NC-T4 are depicted. Results of significance testing are depicted by * for comparison between TV Ad35+ and NC-T4 whereby: * p<0.05, ** p<0.01, *** p<0.001 and **** p<0.0001. § denotes significant differences between TV Ad26+ and NC-T4 whereby: § p<0.05, §§ p<0.01, §§§ p<0.001 and §§§§ p<0.0001. Statistical significance was determined by ANOVA followed by Tukey’s multiple comparisons test where n = 5. B) Multiplex enzyme-linked immunosorbent assay was carried out at week 28 on lung samples as previously described. Results from groups that received immunotherapy (TV Ad35+ and TV Ad26+) and the main control group NC-T4 are shown (mean and standard deviation, n = 4 per group). No statistically significant differences in the levels of IFN-γ, TNF-α or IL-2 were measured between all groups (IFN-γ: p = 0.35; TNF-α: p = 0.98; IL-2: p = 0.09; ANOVA). C) Histopathology of lung samples at week 12 was examined for signs of immunopathology following two doses of immunotherapy with Ad35-TBS. In comparison to the NC-IU animals, the lungs of animals that underwent therapy appeared similar with minimal signs of lesions, necrosis or significant immune infiltration. Immunotherapy did not result in observable immunopathology. Results of histopathology at week 12 are representative of all time points measured (weeks 4, 6, 8 and 10) and at week 18 following Ad26-TBS boosting (data not shown).
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pone.0127907.g007: Probing the mechanism of failure at week 28.A) ELISpot analysis was conducted at the final relapse time point of week 28 as previously described. Results of groups that received immunotherapy (TV Ad35+ and TV Ad26+) and the main negative control group NC-T4 are depicted. Results of significance testing are depicted by * for comparison between TV Ad35+ and NC-T4 whereby: * p<0.05, ** p<0.01, *** p<0.001 and **** p<0.0001. § denotes significant differences between TV Ad26+ and NC-T4 whereby: § p<0.05, §§ p<0.01, §§§ p<0.001 and §§§§ p<0.0001. Statistical significance was determined by ANOVA followed by Tukey’s multiple comparisons test where n = 5. B) Multiplex enzyme-linked immunosorbent assay was carried out at week 28 on lung samples as previously described. Results from groups that received immunotherapy (TV Ad35+ and TV Ad26+) and the main control group NC-T4 are shown (mean and standard deviation, n = 4 per group). No statistically significant differences in the levels of IFN-γ, TNF-α or IL-2 were measured between all groups (IFN-γ: p = 0.35; TNF-α: p = 0.98; IL-2: p = 0.09; ANOVA). C) Histopathology of lung samples at week 12 was examined for signs of immunopathology following two doses of immunotherapy with Ad35-TBS. In comparison to the NC-IU animals, the lungs of animals that underwent therapy appeared similar with minimal signs of lesions, necrosis or significant immune infiltration. Immunotherapy did not result in observable immunopathology. Results of histopathology at week 12 are representative of all time points measured (weeks 4, 6, 8 and 10) and at week 18 following Ad26-TBS boosting (data not shown).

Mentions: We examined the state of the immune system at week 28, to explore possible explanations for the failure of immunotherapy in preventing relapse. Analysis of splenocyte IFN-γ production to pooled peptides revealed significant elevation of immune responses in the immunotherapy groups TV Ad35+ and TV Ad26+ to Ag85A and Ag85B compared to the NC-T4 control but this difference was not seen in antigen TB10.4 responses following Ad35-TBS vaccination but only after Ad26-TBS boosting (Fig 7a). Whilst the levels of Ag85A CD4+ IFN-γ production in the TV Ad35+ and TV Ad26+ animals at week 28 did not defer considerably from the last measurements taken at weeks 16 and 18 respectively, we noticed an increase in the amount of CD4+ T-cells producing IFN-γ in the NC-T4 control group between weeks 10 to 28 (p<0.01, t-test). This ‘catch-up’ response in the control group resulted in no distinguishable differences in CD4 Ag85A IFN-γ values between TV Ad35+ and the control group NC-T4, although boosting with Ad26-TBS did increase the responses further (p<0.0001; ANOVA) (Fig 7a). On the contrary, the level of Ag85A CD8+ IFN-γ production was markedly higher in the TV Ad35+ and TV Ad26+ animals as opposed to the NC-T4 control (TV Ad35+: p<0.0001; TV Ad26+: p<0.0001; ANOVA) (Fig 7a). These results demonstrate that immunotherapy with Ad35-TBS induced both CD4+ and CD8+ responses to Ag85A early in treatment, but only CD8+ responses remained significantly elevated compared to the NC-T4 control by the end of the study. Boosting with Ad26-TBS helped to heighten T-cell responses to all vaccine antigens further by the final time point at week 28.


Immunogenicity without Efficacy of an Adenoviral Tuberculosis Vaccine in a Stringent Mouse Model for Immunotherapy during Treatment.

Alyahya SA, Nolan ST, Smith CM, Bishai WR, Sadoff J, Lamichhane G - PLoS ONE (2015)

Probing the mechanism of failure at week 28.A) ELISpot analysis was conducted at the final relapse time point of week 28 as previously described. Results of groups that received immunotherapy (TV Ad35+ and TV Ad26+) and the main negative control group NC-T4 are depicted. Results of significance testing are depicted by * for comparison between TV Ad35+ and NC-T4 whereby: * p<0.05, ** p<0.01, *** p<0.001 and **** p<0.0001. § denotes significant differences between TV Ad26+ and NC-T4 whereby: § p<0.05, §§ p<0.01, §§§ p<0.001 and §§§§ p<0.0001. Statistical significance was determined by ANOVA followed by Tukey’s multiple comparisons test where n = 5. B) Multiplex enzyme-linked immunosorbent assay was carried out at week 28 on lung samples as previously described. Results from groups that received immunotherapy (TV Ad35+ and TV Ad26+) and the main control group NC-T4 are shown (mean and standard deviation, n = 4 per group). No statistically significant differences in the levels of IFN-γ, TNF-α or IL-2 were measured between all groups (IFN-γ: p = 0.35; TNF-α: p = 0.98; IL-2: p = 0.09; ANOVA). C) Histopathology of lung samples at week 12 was examined for signs of immunopathology following two doses of immunotherapy with Ad35-TBS. In comparison to the NC-IU animals, the lungs of animals that underwent therapy appeared similar with minimal signs of lesions, necrosis or significant immune infiltration. Immunotherapy did not result in observable immunopathology. Results of histopathology at week 12 are representative of all time points measured (weeks 4, 6, 8 and 10) and at week 18 following Ad26-TBS boosting (data not shown).
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pone.0127907.g007: Probing the mechanism of failure at week 28.A) ELISpot analysis was conducted at the final relapse time point of week 28 as previously described. Results of groups that received immunotherapy (TV Ad35+ and TV Ad26+) and the main negative control group NC-T4 are depicted. Results of significance testing are depicted by * for comparison between TV Ad35+ and NC-T4 whereby: * p<0.05, ** p<0.01, *** p<0.001 and **** p<0.0001. § denotes significant differences between TV Ad26+ and NC-T4 whereby: § p<0.05, §§ p<0.01, §§§ p<0.001 and §§§§ p<0.0001. Statistical significance was determined by ANOVA followed by Tukey’s multiple comparisons test where n = 5. B) Multiplex enzyme-linked immunosorbent assay was carried out at week 28 on lung samples as previously described. Results from groups that received immunotherapy (TV Ad35+ and TV Ad26+) and the main control group NC-T4 are shown (mean and standard deviation, n = 4 per group). No statistically significant differences in the levels of IFN-γ, TNF-α or IL-2 were measured between all groups (IFN-γ: p = 0.35; TNF-α: p = 0.98; IL-2: p = 0.09; ANOVA). C) Histopathology of lung samples at week 12 was examined for signs of immunopathology following two doses of immunotherapy with Ad35-TBS. In comparison to the NC-IU animals, the lungs of animals that underwent therapy appeared similar with minimal signs of lesions, necrosis or significant immune infiltration. Immunotherapy did not result in observable immunopathology. Results of histopathology at week 12 are representative of all time points measured (weeks 4, 6, 8 and 10) and at week 18 following Ad26-TBS boosting (data not shown).
Mentions: We examined the state of the immune system at week 28, to explore possible explanations for the failure of immunotherapy in preventing relapse. Analysis of splenocyte IFN-γ production to pooled peptides revealed significant elevation of immune responses in the immunotherapy groups TV Ad35+ and TV Ad26+ to Ag85A and Ag85B compared to the NC-T4 control but this difference was not seen in antigen TB10.4 responses following Ad35-TBS vaccination but only after Ad26-TBS boosting (Fig 7a). Whilst the levels of Ag85A CD4+ IFN-γ production in the TV Ad35+ and TV Ad26+ animals at week 28 did not defer considerably from the last measurements taken at weeks 16 and 18 respectively, we noticed an increase in the amount of CD4+ T-cells producing IFN-γ in the NC-T4 control group between weeks 10 to 28 (p<0.01, t-test). This ‘catch-up’ response in the control group resulted in no distinguishable differences in CD4 Ag85A IFN-γ values between TV Ad35+ and the control group NC-T4, although boosting with Ad26-TBS did increase the responses further (p<0.0001; ANOVA) (Fig 7a). On the contrary, the level of Ag85A CD8+ IFN-γ production was markedly higher in the TV Ad35+ and TV Ad26+ animals as opposed to the NC-T4 control (TV Ad35+: p<0.0001; TV Ad26+: p<0.0001; ANOVA) (Fig 7a). These results demonstrate that immunotherapy with Ad35-TBS induced both CD4+ and CD8+ responses to Ag85A early in treatment, but only CD8+ responses remained significantly elevated compared to the NC-T4 control by the end of the study. Boosting with Ad26-TBS helped to heighten T-cell responses to all vaccine antigens further by the final time point at week 28.

Bottom Line: Even though immunotherapy resulted in strong splenic IFN-γ responses, no effect on bacterial replication in the lungs was seen.Multiplex assay analysis of lung samples revealed the absence of cytokine augmentation such as IFN-γ, TNF-α and IL-2, suggesting that immunization failed to induce immunity in the lungs.The results obtained from our study raise questions regarding the traits of protective TB immunity that are relevant for the development of future immunotherapeutic and post-exposure vaccination strategies.

View Article: PubMed Central - PubMed

Affiliation: Crucell Holland B.V., Janssen Infectious Diseases and Vaccines, Leiden, The Netherlands.

ABSTRACT
To investigate if bacterial persistence during TB drug treatment could be overcome by modulation of host immunity, we adapted a clinically-relevant model developed for the evaluation of new drugs and examined if immunotherapy with two adenoviral vaccines, Ad35-TBS (AERAS-402) and Ad26-TBS, could shorten therapy in mice. Even though immunotherapy resulted in strong splenic IFN-γ responses, no effect on bacterial replication in the lungs was seen. Multiplex assay analysis of lung samples revealed the absence of cytokine augmentation such as IFN-γ, TNF-α and IL-2, suggesting that immunization failed to induce immunity in the lungs. In this model, we show that IFN-γ levels were not associated with protection against disease relapse. The results obtained from our study raise questions regarding the traits of protective TB immunity that are relevant for the development of future immunotherapeutic and post-exposure vaccination strategies.

No MeSH data available.


Related in: MedlinePlus