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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

Safety assessment of the Qβ-mIL-1b(D143K) vaccine in mice. (a) Time course of anti-IL-1β antibody responses. Female C57BL/6 mice (n = 4) were immunized s.c. on days 0, 14, and 35 (arrows) with 1 µg of Qβ-mIL-1b(D143K). On days 42, 70, 140, and 228, mice were challenged with i.p. injections of 1 µg wild-type mouse IL-1β. Three hours after challenge, sera were collected, and IL-6 levels were quantified by ELISA. Single data points (filled gray circles) represent individual animals. Antimouse IL-1β (wild type) IgG antibody titers were determined at the same time points by ELISA and are represented as group means ± SEM (black triangles). (b) Effect of increased IL-1β levels on antimouse IL-1β IgG antibody titers. Groups of female C57BL/6 mice (n = 5) were immunized s.c. on day 0 with 50 µg Qβ-mIL-1b(D143K). On day 76, mice received either an i.v. injection of 100 ng wild-type mouse IL-1β (in 100 µl PBS), an i.p. injection of a mixture of 1 ng Escherichia coli lipopolysaccharide, and 20 mg N-galactosamine (Sigma-Aldrich), or a s.c. injection of 50 µg Qβ-mIL-1b(D143K). Control groups received either an i.v. injection of 100 µl PBS or a s.c. injection of 50 µg Qβ VLPs. Mice were bled on days 69, 76, 83, and 90 and mouse IL-1β (wild type)-specific IgG antibody titers were determined by ELISA. Shown are group means ± SEM. (c) Measurement of IL-1β-specific T cell responses after vaccination with Qβ-mIL-1b(D143K). Groups of mice were immunized with Qβ-mIL-1b(D143K)-p13 in the presence of CpG or Alum, respectively, as described in Materials and Methods. One group of female C57BL/6 mice was kept naive. After immunization, splenocytes were isolated from all mice and stimulated with BMDC that had been loaded either with synthetic p13 peptide or with wild-type mouse IL-1β. Nonspecific IFNγ-release from CD4+ T cells was determined by incubation of splenocytes with mock-pulsed BMDC (no antigen). Antigen-specific IFNγ-producing CD4+ T cells were determined by fluorescence-activated cell sorting. Shown are group means ± SEM. (d) Th cell dependence of anti-IL-1β antibody induction. Groups of mice received s.c. injections of either wild-type mouse IL-1β or mIL-1b(D143K)-p13, each in the presence of incomplete Freund’s adjuvant. A subgroup of mIL-1b(D143K)-p13-immunized mice were additionally injected with a depleting anti-CD4 antibody. Mouse IL-1β (wild type)-specific IgG antibody titers were analyzed by ELISA. Shown are group means ± SEM.
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fig3: Safety assessment of the Qβ-mIL-1b(D143K) vaccine in mice. (a) Time course of anti-IL-1β antibody responses. Female C57BL/6 mice (n = 4) were immunized s.c. on days 0, 14, and 35 (arrows) with 1 µg of Qβ-mIL-1b(D143K). On days 42, 70, 140, and 228, mice were challenged with i.p. injections of 1 µg wild-type mouse IL-1β. Three hours after challenge, sera were collected, and IL-6 levels were quantified by ELISA. Single data points (filled gray circles) represent individual animals. Antimouse IL-1β (wild type) IgG antibody titers were determined at the same time points by ELISA and are represented as group means ± SEM (black triangles). (b) Effect of increased IL-1β levels on antimouse IL-1β IgG antibody titers. Groups of female C57BL/6 mice (n = 5) were immunized s.c. on day 0 with 50 µg Qβ-mIL-1b(D143K). On day 76, mice received either an i.v. injection of 100 ng wild-type mouse IL-1β (in 100 µl PBS), an i.p. injection of a mixture of 1 ng Escherichia coli lipopolysaccharide, and 20 mg N-galactosamine (Sigma-Aldrich), or a s.c. injection of 50 µg Qβ-mIL-1b(D143K). Control groups received either an i.v. injection of 100 µl PBS or a s.c. injection of 50 µg Qβ VLPs. Mice were bled on days 69, 76, 83, and 90 and mouse IL-1β (wild type)-specific IgG antibody titers were determined by ELISA. Shown are group means ± SEM. (c) Measurement of IL-1β-specific T cell responses after vaccination with Qβ-mIL-1b(D143K). Groups of mice were immunized with Qβ-mIL-1b(D143K)-p13 in the presence of CpG or Alum, respectively, as described in Materials and Methods. One group of female C57BL/6 mice was kept naive. After immunization, splenocytes were isolated from all mice and stimulated with BMDC that had been loaded either with synthetic p13 peptide or with wild-type mouse IL-1β. Nonspecific IFNγ-release from CD4+ T cells was determined by incubation of splenocytes with mock-pulsed BMDC (no antigen). Antigen-specific IFNγ-producing CD4+ T cells were determined by fluorescence-activated cell sorting. Shown are group means ± SEM. (d) Th cell dependence of anti-IL-1β antibody induction. Groups of mice received s.c. injections of either wild-type mouse IL-1β or mIL-1b(D143K)-p13, each in the presence of incomplete Freund’s adjuvant. A subgroup of mIL-1b(D143K)-p13-immunized mice were additionally injected with a depleting anti-CD4 antibody. Mouse IL-1β (wild type)-specific IgG antibody titers were analyzed by ELISA. Shown are group means ± SEM.

Mentions: A possible safety concern for the use of a vaccine against IL-1β in humans is uncontrolled, irreversible neutralization of the cytokine through the induction of long-lasting or even steadily increasing titers of IL-1β-specific antibodies. Such persisting anti-IL-β antibody titers could have long-term consequences for immunity against infectious diseases. To test for this possibility in a preclinical setting, we first immunized mice three times with Qβ-mIL-1b(D143K) to induce a robust mIL-1β-neutralizing antibody response. Starting from the peak of the antibody response, we then followed the mIL-1β-specific antibody titers over an extended time period while measuring the neutralizing activity of the antibodies in vivo by repeated challenges with a constant dose of recombinant wild-type mIL-1β. On day 42, at the peak of the antibody response, anti-mIL-1β IgG titers of ~27,900 were measured, and an i.p. challenge with 1 µg of mIL-1β did not induce any elevation of serum IL-6 levels, indicating complete neutralization of the applied dose of mIL-1β by vaccine-induced antibodies (Figure 3a). During the follow-up phase, antibody titers continuously declined and responsiveness to the challenge was gradually regained. On day 228, three out of four mice produced robust amounts of IL-6 in response to injection with mIL-1β. Importantly, the IL-1β-specific antibody response was not boosted by the repeated administration of recombinant wild-type IL-1β, indicating that nonconjugated IL-1β is unable to activate IL-1β-specific B cells induced by vaccination.


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)

Safety assessment of the Qβ-mIL-1b(D143K) vaccine in mice. (a) Time course of anti-IL-1β antibody responses. Female C57BL/6 mice (n = 4) were immunized s.c. on days 0, 14, and 35 (arrows) with 1 µg of Qβ-mIL-1b(D143K). On days 42, 70, 140, and 228, mice were challenged with i.p. injections of 1 µg wild-type mouse IL-1β. Three hours after challenge, sera were collected, and IL-6 levels were quantified by ELISA. Single data points (filled gray circles) represent individual animals. Antimouse IL-1β (wild type) IgG antibody titers were determined at the same time points by ELISA and are represented as group means ± SEM (black triangles). (b) Effect of increased IL-1β levels on antimouse IL-1β IgG antibody titers. Groups of female C57BL/6 mice (n = 5) were immunized s.c. on day 0 with 50 µg Qβ-mIL-1b(D143K). On day 76, mice received either an i.v. injection of 100 ng wild-type mouse IL-1β (in 100 µl PBS), an i.p. injection of a mixture of 1 ng Escherichia coli lipopolysaccharide, and 20 mg N-galactosamine (Sigma-Aldrich), or a s.c. injection of 50 µg Qβ-mIL-1b(D143K). Control groups received either an i.v. injection of 100 µl PBS or a s.c. injection of 50 µg Qβ VLPs. Mice were bled on days 69, 76, 83, and 90 and mouse IL-1β (wild type)-specific IgG antibody titers were determined by ELISA. Shown are group means ± SEM. (c) Measurement of IL-1β-specific T cell responses after vaccination with Qβ-mIL-1b(D143K). Groups of mice were immunized with Qβ-mIL-1b(D143K)-p13 in the presence of CpG or Alum, respectively, as described in Materials and Methods. One group of female C57BL/6 mice was kept naive. After immunization, splenocytes were isolated from all mice and stimulated with BMDC that had been loaded either with synthetic p13 peptide or with wild-type mouse IL-1β. Nonspecific IFNγ-release from CD4+ T cells was determined by incubation of splenocytes with mock-pulsed BMDC (no antigen). Antigen-specific IFNγ-producing CD4+ T cells were determined by fluorescence-activated cell sorting. Shown are group means ± SEM. (d) Th cell dependence of anti-IL-1β antibody induction. Groups of mice received s.c. injections of either wild-type mouse IL-1β or mIL-1b(D143K)-p13, each in the presence of incomplete Freund’s adjuvant. A subgroup of mIL-1b(D143K)-p13-immunized mice were additionally injected with a depleting anti-CD4 antibody. Mouse IL-1β (wild type)-specific IgG antibody titers were analyzed by ELISA. Shown are group means ± SEM.
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Related In: Results  -  Collection

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Show All Figures
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fig3: Safety assessment of the Qβ-mIL-1b(D143K) vaccine in mice. (a) Time course of anti-IL-1β antibody responses. Female C57BL/6 mice (n = 4) were immunized s.c. on days 0, 14, and 35 (arrows) with 1 µg of Qβ-mIL-1b(D143K). On days 42, 70, 140, and 228, mice were challenged with i.p. injections of 1 µg wild-type mouse IL-1β. Three hours after challenge, sera were collected, and IL-6 levels were quantified by ELISA. Single data points (filled gray circles) represent individual animals. Antimouse IL-1β (wild type) IgG antibody titers were determined at the same time points by ELISA and are represented as group means ± SEM (black triangles). (b) Effect of increased IL-1β levels on antimouse IL-1β IgG antibody titers. Groups of female C57BL/6 mice (n = 5) were immunized s.c. on day 0 with 50 µg Qβ-mIL-1b(D143K). On day 76, mice received either an i.v. injection of 100 ng wild-type mouse IL-1β (in 100 µl PBS), an i.p. injection of a mixture of 1 ng Escherichia coli lipopolysaccharide, and 20 mg N-galactosamine (Sigma-Aldrich), or a s.c. injection of 50 µg Qβ-mIL-1b(D143K). Control groups received either an i.v. injection of 100 µl PBS or a s.c. injection of 50 µg Qβ VLPs. Mice were bled on days 69, 76, 83, and 90 and mouse IL-1β (wild type)-specific IgG antibody titers were determined by ELISA. Shown are group means ± SEM. (c) Measurement of IL-1β-specific T cell responses after vaccination with Qβ-mIL-1b(D143K). Groups of mice were immunized with Qβ-mIL-1b(D143K)-p13 in the presence of CpG or Alum, respectively, as described in Materials and Methods. One group of female C57BL/6 mice was kept naive. After immunization, splenocytes were isolated from all mice and stimulated with BMDC that had been loaded either with synthetic p13 peptide or with wild-type mouse IL-1β. Nonspecific IFNγ-release from CD4+ T cells was determined by incubation of splenocytes with mock-pulsed BMDC (no antigen). Antigen-specific IFNγ-producing CD4+ T cells were determined by fluorescence-activated cell sorting. Shown are group means ± SEM. (d) Th cell dependence of anti-IL-1β antibody induction. Groups of mice received s.c. injections of either wild-type mouse IL-1β or mIL-1b(D143K)-p13, each in the presence of incomplete Freund’s adjuvant. A subgroup of mIL-1b(D143K)-p13-immunized mice were additionally injected with a depleting anti-CD4 antibody. Mouse IL-1β (wild type)-specific IgG antibody titers were analyzed by ELISA. Shown are group means ± SEM.
Mentions: A possible safety concern for the use of a vaccine against IL-1β in humans is uncontrolled, irreversible neutralization of the cytokine through the induction of long-lasting or even steadily increasing titers of IL-1β-specific antibodies. Such persisting anti-IL-β antibody titers could have long-term consequences for immunity against infectious diseases. To test for this possibility in a preclinical setting, we first immunized mice three times with Qβ-mIL-1b(D143K) to induce a robust mIL-1β-neutralizing antibody response. Starting from the peak of the antibody response, we then followed the mIL-1β-specific antibody titers over an extended time period while measuring the neutralizing activity of the antibodies in vivo by repeated challenges with a constant dose of recombinant wild-type mIL-1β. On day 42, at the peak of the antibody response, anti-mIL-1β IgG titers of ~27,900 were measured, and an i.p. challenge with 1 µg of mIL-1β did not induce any elevation of serum IL-6 levels, indicating complete neutralization of the applied dose of mIL-1β by vaccine-induced antibodies (Figure 3a). During the follow-up phase, antibody titers continuously declined and responsiveness to the challenge was gradually regained. On day 228, three out of four mice produced robust amounts of IL-6 in response to injection with mIL-1β. Importantly, the IL-1β-specific antibody response was not boosted by the repeated administration of recombinant wild-type IL-1β, indicating that nonconjugated IL-1β is unable to activate IL-1β-specific B cells induced by vaccination.

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