Limits...
Preclinical efficacy and safety of an anti-IL-1β vaccine for the treatment of type 2 diabetes.

Spohn G, Schori C, Keller I, Sladko K, Sina C, Guler R, Schwarz K, Johansen P, Jennings GT, Bachmann MF - Mol Ther Methods Clin Dev (2014)

Bottom Line: Here, we describe the preclinical development of a therapeutic vaccine against IL-1β consisting of a detoxified version of IL-1β chemically cross-linked to virus-like particles of the bacteriophage Qβ.Antibody titers were long lasting but reversible over time and not associated with the development of potentially harmful T cell responses against IL-1β.Hence, immunization with IL-1β conjugated to virus-like particles has the potential to become a safe, efficacious, and cost-effective therapy for the prevention and long-term treatment of type 2 diabetes.

View Article: PubMed Central - PubMed

Affiliation: Cytos Biotechnology , Schlieren, Switzerland.

ABSTRACT
Neutralization of the inflammatory cytokine interleukin-1β (IL-1β) is a promising new strategy to prevent the β-cell destruction, which leads to type 2 diabetes. Here, we describe the preclinical development of a therapeutic vaccine against IL-1β consisting of a detoxified version of IL-1β chemically cross-linked to virus-like particles of the bacteriophage Qβ. The vaccine was well tolerated and induced robust antibody responses in mice, which neutralized the biological activity of IL-1β, as shown both in cellular assays and in challenge experiments in vivo. Antibody titers were long lasting but reversible over time and not associated with the development of potentially harmful T cell responses against IL-1β. Neutralization of IL-1β by vaccine-induced antibodies had no influence on the immune responses of mice to Listeria monocytogenes and Mycobacterium tuberculosis. In a diet-induced model of type 2 diabetes, immunized mice showed improved glucose tolerance, which was mediated by improved insulin secretion by pancreatic β-cells. Hence, immunization with IL-1β conjugated to virus-like particles has the potential to become a safe, efficacious, and cost-effective therapy for the prevention and long-term treatment of type 2 diabetes.

No MeSH data available.


Related in: MedlinePlus

Immunogenicity of Qβ-mIL-1b(D143K) in mice. (a) Induction of anti-mIL-1β antibody titers. Female C57BL/6 mice (n = 5) were immunized twice (days 0 and 42, arrows) with 10 µg of Qβ-mIL-1b(D143K) in the presence of Alum. At the indicated time points, mouse IL-1β (wild type)-specific IgG antibody titers were measured by ELISA (filled black circles). At the same time points IL-1β neutralizing titers were determined ex vivo in the HeLa IL-6 secretion assay as described in Materials and Methods (open gray circles). Shown are mean titers ± SEM. (b) Ratio of IL-1β neutralizing versus ELISA titers over time. The ratios of IL-1β- neutralizing versus IL-1β-specific IgG ELISA titers were calculated for the group means of each time point of the experiment described in a. Vaccine injections are indicated by arrows. (c) Antibody responses in IL-1β-deficient mice. Groups (n = 4) of female C57BL/6 IL-1β-deficient mice (-/-) or C57BL/6 control mice (+/+) were immunized on days 0, 14, and 28 with 10 µg of Qβ-mIL-1b(D143K) in the presence of Alum. On day 42, mouse IL-1β (wild type)- as well as Qβ-specific IgG antibody titers were measured by ELISA. Shown are mean titers ± SEM. (d) Induction of IL-1β-neutralizing titers in IL-1β-deficient mice. Mouse IL-1β (wild type)-neutralizing titers were determined in sera from day 42 of the experiment described in c. Shown are mean titers ± SEM. (e) In vivo neutralization of IL-1β in Qβ-mIL-1b(D143K)-immunized mice. Groups of female C57BL/6 mice (n = 5) were immunized s.c. on days 0, 14, and 28 with 1 µg of Qβ-mIL-1b(D143K) or Qβ VLPs as control. On day 42, mice were challenged with an i.p. injection of 1 µg wild-type mouse IL-1β. Three hours after challenge, sera were collected and IL-6 levels were quantified with a Quantikine ELISA kit. Shown are data from individual mice and group means (*P < 0.001; n.s., not significant).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4362373&req=5

fig2: Immunogenicity of Qβ-mIL-1b(D143K) in mice. (a) Induction of anti-mIL-1β antibody titers. Female C57BL/6 mice (n = 5) were immunized twice (days 0 and 42, arrows) with 10 µg of Qβ-mIL-1b(D143K) in the presence of Alum. At the indicated time points, mouse IL-1β (wild type)-specific IgG antibody titers were measured by ELISA (filled black circles). At the same time points IL-1β neutralizing titers were determined ex vivo in the HeLa IL-6 secretion assay as described in Materials and Methods (open gray circles). Shown are mean titers ± SEM. (b) Ratio of IL-1β neutralizing versus ELISA titers over time. The ratios of IL-1β- neutralizing versus IL-1β-specific IgG ELISA titers were calculated for the group means of each time point of the experiment described in a. Vaccine injections are indicated by arrows. (c) Antibody responses in IL-1β-deficient mice. Groups (n = 4) of female C57BL/6 IL-1β-deficient mice (-/-) or C57BL/6 control mice (+/+) were immunized on days 0, 14, and 28 with 10 µg of Qβ-mIL-1b(D143K) in the presence of Alum. On day 42, mouse IL-1β (wild type)- as well as Qβ-specific IgG antibody titers were measured by ELISA. Shown are mean titers ± SEM. (d) Induction of IL-1β-neutralizing titers in IL-1β-deficient mice. Mouse IL-1β (wild type)-neutralizing titers were determined in sera from day 42 of the experiment described in c. Shown are mean titers ± SEM. (e) In vivo neutralization of IL-1β in Qβ-mIL-1b(D143K)-immunized mice. Groups of female C57BL/6 mice (n = 5) were immunized s.c. on days 0, 14, and 28 with 1 µg of Qβ-mIL-1b(D143K) or Qβ VLPs as control. On day 42, mice were challenged with an i.p. injection of 1 µg wild-type mouse IL-1β. Three hours after challenge, sera were collected and IL-6 levels were quantified with a Quantikine ELISA kit. Shown are data from individual mice and group means (*P < 0.001; n.s., not significant).

Mentions: To enable preclinical testing of the anti-IL-1β vaccine in the context of self-specific immunization, we produced the murine version of hIL-1b(D145K) (=mIL-1b(D143K)) and conjugated it to Qβ VLPs. Figure 2a shows that a single injection of the conjugate vaccine Qβ-mIL-1b(D143K) induced antimurine IL-1β (mIL-1β) IgG titers of 17,600 by day 21, which increased to 33,900 by day 42. An additional injection on day 42 led to a further increase in titers to a maximum of 65,200 by day 70. In the absence of further injections, the titers then declined with an approximate half-life of 2–3 months. Moderate mIL-1β-neutralizing titers of 50 were detected in sera from day 21, which increased to 300 on day 42 and reached their maximum of 1,200 on day 70. Thereafter, they declined in parallel to the enzyme-linked immunosorbent assay (ELISA) titers until the end of the observation period. The ratio of mIL-1β-specific neutralization versus ELISA titers (Figure 2b) increased from day 21 to 70, indicating an augmentation in the average affinity of the produced anti-IL-1β antibodies over time. To assess if the magnitude and quality of the anti-mIL-1β antibody response is affected by the repertoire of IL-1β-specific B cells, we compared wild-type and IL-1β-deficient mice. If the presence of IL-1β in wild-type animals causes a certain degree of B cell tolerance toward IL-1β as an antigen, one would expect that the Qβ-mIL-1b(D143K) vaccine induces stronger IL-1β-specific B cell responses in IL-1β-deficient mice due to the absence of B cell tolerance and thus a larger number and a broader repertoire of IL-1β-specific B cells. Groups of IL-1β-deficient and wild-type control mice were immunized with Qβ-mIL-1b(D143K), and antibody responses against mIL-1β and the Qβ VLP carrier were analyzed by ELISA. As shown in Figure 2c, IgG titers against the foreign antigen Qβ were not significantly reduced in mutant mice, indicating that IL-1β deficiency does not lead to a general suppression of the humoral response. Importantly, both ELISA titers against mIL-1β (Figure 2c) as well as IL-1β-neutralizing titers (Figure 2d) were similar between wild-type and IL-1β-deficient mice, suggesting that no obvious restriction of the IL-1β-specific B cell repertoire is present in wild-type mice.


Preclinical efficacy and safety of an anti-IL-1β vaccine for the treatment of type 2 diabetes.

Spohn G, Schori C, Keller I, Sladko K, Sina C, Guler R, Schwarz K, Johansen P, Jennings GT, Bachmann MF - Mol Ther Methods Clin Dev (2014)

Immunogenicity of Qβ-mIL-1b(D143K) in mice. (a) Induction of anti-mIL-1β antibody titers. Female C57BL/6 mice (n = 5) were immunized twice (days 0 and 42, arrows) with 10 µg of Qβ-mIL-1b(D143K) in the presence of Alum. At the indicated time points, mouse IL-1β (wild type)-specific IgG antibody titers were measured by ELISA (filled black circles). At the same time points IL-1β neutralizing titers were determined ex vivo in the HeLa IL-6 secretion assay as described in Materials and Methods (open gray circles). Shown are mean titers ± SEM. (b) Ratio of IL-1β neutralizing versus ELISA titers over time. The ratios of IL-1β- neutralizing versus IL-1β-specific IgG ELISA titers were calculated for the group means of each time point of the experiment described in a. Vaccine injections are indicated by arrows. (c) Antibody responses in IL-1β-deficient mice. Groups (n = 4) of female C57BL/6 IL-1β-deficient mice (-/-) or C57BL/6 control mice (+/+) were immunized on days 0, 14, and 28 with 10 µg of Qβ-mIL-1b(D143K) in the presence of Alum. On day 42, mouse IL-1β (wild type)- as well as Qβ-specific IgG antibody titers were measured by ELISA. Shown are mean titers ± SEM. (d) Induction of IL-1β-neutralizing titers in IL-1β-deficient mice. Mouse IL-1β (wild type)-neutralizing titers were determined in sera from day 42 of the experiment described in c. Shown are mean titers ± SEM. (e) In vivo neutralization of IL-1β in Qβ-mIL-1b(D143K)-immunized mice. Groups of female C57BL/6 mice (n = 5) were immunized s.c. on days 0, 14, and 28 with 1 µg of Qβ-mIL-1b(D143K) or Qβ VLPs as control. On day 42, mice were challenged with an i.p. injection of 1 µg wild-type mouse IL-1β. Three hours after challenge, sera were collected and IL-6 levels were quantified with a Quantikine ELISA kit. Shown are data from individual mice and group means (*P < 0.001; n.s., not significant).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362373&req=5

fig2: Immunogenicity of Qβ-mIL-1b(D143K) in mice. (a) Induction of anti-mIL-1β antibody titers. Female C57BL/6 mice (n = 5) were immunized twice (days 0 and 42, arrows) with 10 µg of Qβ-mIL-1b(D143K) in the presence of Alum. At the indicated time points, mouse IL-1β (wild type)-specific IgG antibody titers were measured by ELISA (filled black circles). At the same time points IL-1β neutralizing titers were determined ex vivo in the HeLa IL-6 secretion assay as described in Materials and Methods (open gray circles). Shown are mean titers ± SEM. (b) Ratio of IL-1β neutralizing versus ELISA titers over time. The ratios of IL-1β- neutralizing versus IL-1β-specific IgG ELISA titers were calculated for the group means of each time point of the experiment described in a. Vaccine injections are indicated by arrows. (c) Antibody responses in IL-1β-deficient mice. Groups (n = 4) of female C57BL/6 IL-1β-deficient mice (-/-) or C57BL/6 control mice (+/+) were immunized on days 0, 14, and 28 with 10 µg of Qβ-mIL-1b(D143K) in the presence of Alum. On day 42, mouse IL-1β (wild type)- as well as Qβ-specific IgG antibody titers were measured by ELISA. Shown are mean titers ± SEM. (d) Induction of IL-1β-neutralizing titers in IL-1β-deficient mice. Mouse IL-1β (wild type)-neutralizing titers were determined in sera from day 42 of the experiment described in c. Shown are mean titers ± SEM. (e) In vivo neutralization of IL-1β in Qβ-mIL-1b(D143K)-immunized mice. Groups of female C57BL/6 mice (n = 5) were immunized s.c. on days 0, 14, and 28 with 1 µg of Qβ-mIL-1b(D143K) or Qβ VLPs as control. On day 42, mice were challenged with an i.p. injection of 1 µg wild-type mouse IL-1β. Three hours after challenge, sera were collected and IL-6 levels were quantified with a Quantikine ELISA kit. Shown are data from individual mice and group means (*P < 0.001; n.s., not significant).
Mentions: To enable preclinical testing of the anti-IL-1β vaccine in the context of self-specific immunization, we produced the murine version of hIL-1b(D145K) (=mIL-1b(D143K)) and conjugated it to Qβ VLPs. Figure 2a shows that a single injection of the conjugate vaccine Qβ-mIL-1b(D143K) induced antimurine IL-1β (mIL-1β) IgG titers of 17,600 by day 21, which increased to 33,900 by day 42. An additional injection on day 42 led to a further increase in titers to a maximum of 65,200 by day 70. In the absence of further injections, the titers then declined with an approximate half-life of 2–3 months. Moderate mIL-1β-neutralizing titers of 50 were detected in sera from day 21, which increased to 300 on day 42 and reached their maximum of 1,200 on day 70. Thereafter, they declined in parallel to the enzyme-linked immunosorbent assay (ELISA) titers until the end of the observation period. The ratio of mIL-1β-specific neutralization versus ELISA titers (Figure 2b) increased from day 21 to 70, indicating an augmentation in the average affinity of the produced anti-IL-1β antibodies over time. To assess if the magnitude and quality of the anti-mIL-1β antibody response is affected by the repertoire of IL-1β-specific B cells, we compared wild-type and IL-1β-deficient mice. If the presence of IL-1β in wild-type animals causes a certain degree of B cell tolerance toward IL-1β as an antigen, one would expect that the Qβ-mIL-1b(D143K) vaccine induces stronger IL-1β-specific B cell responses in IL-1β-deficient mice due to the absence of B cell tolerance and thus a larger number and a broader repertoire of IL-1β-specific B cells. Groups of IL-1β-deficient and wild-type control mice were immunized with Qβ-mIL-1b(D143K), and antibody responses against mIL-1β and the Qβ VLP carrier were analyzed by ELISA. As shown in Figure 2c, IgG titers against the foreign antigen Qβ were not significantly reduced in mutant mice, indicating that IL-1β deficiency does not lead to a general suppression of the humoral response. Importantly, both ELISA titers against mIL-1β (Figure 2c) as well as IL-1β-neutralizing titers (Figure 2d) were similar between wild-type and IL-1β-deficient mice, suggesting that no obvious restriction of the IL-1β-specific B cell repertoire is present in wild-type mice.

Bottom Line: Here, we describe the preclinical development of a therapeutic vaccine against IL-1β consisting of a detoxified version of IL-1β chemically cross-linked to virus-like particles of the bacteriophage Qβ.Antibody titers were long lasting but reversible over time and not associated with the development of potentially harmful T cell responses against IL-1β.Hence, immunization with IL-1β conjugated to virus-like particles has the potential to become a safe, efficacious, and cost-effective therapy for the prevention and long-term treatment of type 2 diabetes.

View Article: PubMed Central - PubMed

Affiliation: Cytos Biotechnology , Schlieren, Switzerland.

ABSTRACT
Neutralization of the inflammatory cytokine interleukin-1β (IL-1β) is a promising new strategy to prevent the β-cell destruction, which leads to type 2 diabetes. Here, we describe the preclinical development of a therapeutic vaccine against IL-1β consisting of a detoxified version of IL-1β chemically cross-linked to virus-like particles of the bacteriophage Qβ. The vaccine was well tolerated and induced robust antibody responses in mice, which neutralized the biological activity of IL-1β, as shown both in cellular assays and in challenge experiments in vivo. Antibody titers were long lasting but reversible over time and not associated with the development of potentially harmful T cell responses against IL-1β. Neutralization of IL-1β by vaccine-induced antibodies had no influence on the immune responses of mice to Listeria monocytogenes and Mycobacterium tuberculosis. In a diet-induced model of type 2 diabetes, immunized mice showed improved glucose tolerance, which was mediated by improved insulin secretion by pancreatic β-cells. Hence, immunization with IL-1β conjugated to virus-like particles has the potential to become a safe, efficacious, and cost-effective therapy for the prevention and long-term treatment of type 2 diabetes.

No MeSH data available.


Related in: MedlinePlus