Limits...
Virtual optimization of nasal insulin therapy predicts immunization frequency to be crucial for diabetes protection.

Fousteri G, Chan JR, Zheng Y, Whiting C, Dave A, Bresson D, Croft M, von Herrath M - Diabetes (2010)

Bottom Line: The experimental aim was to evaluate the impact of dose, frequency of administration, and age at treatment on Treg induction and optimal therapeutic outcome.Here, the advantage of applying computer modeling in optimizing the therapeutic efficacy of nasal insulin immunotherapy was confirmed.In silico modeling was able to streamline the experimental design and to identify the particular time frame at which biomarkers associated with protection in live NODs were induced.

View Article: PubMed Central - PubMed

Affiliation: Diabetes Center, La Jolla Institute for Allergy and Immunology, La Jolla, California, USA.

ABSTRACT

Objective: Development of antigen-specific strategies to treat or prevent type 1 diabetes has been slow and difficult because of the lack of experimental tools and defined biomarkers that account for the underlying therapeutic mechanisms.

Research design and methods: The type 1 diabetes PhysioLab platform, a large-scale mathematical model of disease pathogenesis in the nonobese diabetic (NOD) mouse, was used to investigate the possible mechanisms underlying the efficacy of nasal insulin B:9-23 peptide therapy. The experimental aim was to evaluate the impact of dose, frequency of administration, and age at treatment on Treg induction and optimal therapeutic outcome.

Results: In virtual NOD mice, treatment efficacy was predicted to depend primarily on the immunization frequency and stage of the disease and to a lesser extent on the dose. Whereas low-frequency immunization protected from diabetes atrributed to Treg and interleukin (IL)-10 induction in the pancreas 1-2 weeks after treatment, high-frequency immunization failed. These predictions were confirmed with wet-lab approaches, where only low-frequency immunization started at an early disease stage in the NOD mouse resulted in significant protection from diabetes by inducing IL-10 and Treg.

Conclusions: Here, the advantage of applying computer modeling in optimizing the therapeutic efficacy of nasal insulin immunotherapy was confirmed. In silico modeling was able to streamline the experimental design and to identify the particular time frame at which biomarkers associated with protection in live NODs were induced. These results support the development and application of humanized platforms for the design of clinical trials (i.e., for the ongoing nasal insulin prevention studies).

Show MeSH

Related in: MedlinePlus

Nasal B:9–23 administration started at 4 weeks of age upon low-frequency immunization decreases insulitis and increases Treg in the islets. A–C: Reduced insulitis in low-frequency B:9-23 nasal immunized NOD. A: Pancreatic sections from B:9-23 nasally treated and control (PBS) mice were scored as described in research design and methods. The percentages represent the number of islets of a given score over the total number of islets examined. Scoring was performed on histological sections derived from 12-week-old nondiabetic mice (n = 4 mice per group). B: Pancreatic histology. Pancreatic sections (6 μm thick) were stained for insulin (blue) and CD8 (red) (C) or CD4 (red) (D) and analyzed at ×10 magnification. All histological sections shown were derived from 12-week-old nondiabetic mice. D–F: Increased Treg numbers in the islets of B:9-23–immunized mice. D: In silico–predicted changes in Treg levels within the pancreatic islet. The green box indicates the suggested-by-the-model-age window at which the laboratory measurements should be conducted. E: Actual Treg frequency as determined by measuring Foxp3+ cells per CD4+ cells per islet according to degree of insulitis. Same histological sections as in B and C were analyzed after immunofluorescent staining (IFS) for CD4, insulin, and Foxp3. Data are means ± SE. F: Representative microphotograph from each group of mice with insulitis degree 4 is shown at ×20 magnification (IFS for CD4 in blue, insulin in red, and Foxp3 in green). *P < 0.05 between PBS and high-frequency, high-frequency versus low-frequency immunization. **P < 0.01 between PBS and low-frequency immunization. (A high-quality digital representation of this figure is available in the online issue.)
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2992777&req=5

Figure 4: Nasal B:9–23 administration started at 4 weeks of age upon low-frequency immunization decreases insulitis and increases Treg in the islets. A–C: Reduced insulitis in low-frequency B:9-23 nasal immunized NOD. A: Pancreatic sections from B:9-23 nasally treated and control (PBS) mice were scored as described in research design and methods. The percentages represent the number of islets of a given score over the total number of islets examined. Scoring was performed on histological sections derived from 12-week-old nondiabetic mice (n = 4 mice per group). B: Pancreatic histology. Pancreatic sections (6 μm thick) were stained for insulin (blue) and CD8 (red) (C) or CD4 (red) (D) and analyzed at ×10 magnification. All histological sections shown were derived from 12-week-old nondiabetic mice. D–F: Increased Treg numbers in the islets of B:9-23–immunized mice. D: In silico–predicted changes in Treg levels within the pancreatic islet. The green box indicates the suggested-by-the-model-age window at which the laboratory measurements should be conducted. E: Actual Treg frequency as determined by measuring Foxp3+ cells per CD4+ cells per islet according to degree of insulitis. Same histological sections as in B and C were analyzed after immunofluorescent staining (IFS) for CD4, insulin, and Foxp3. Data are means ± SE. F: Representative microphotograph from each group of mice with insulitis degree 4 is shown at ×20 magnification (IFS for CD4 in blue, insulin in red, and Foxp3 in green). *P < 0.05 between PBS and high-frequency, high-frequency versus low-frequency immunization. **P < 0.01 between PBS and low-frequency immunization. (A high-quality digital representation of this figure is available in the online issue.)

Mentions: To determine the extent of islet infiltration following nasal B:9-23 immunization, pancreata from nondiabetic mice of all treatment groups were histologically examined at 12 weeks of age. As shown in Fig. 4A–C, the majority of islets in animals treated with the low-frequency protocol was minimally inflamed or had mild peri-insulitis. In contrast, most of the islets of normoglycemic mice that received the high-frequency immunization protocol had much greater CD4+ and CD8+ T-cell infiltrates. Furthermore, when scoring of the islets was undertaken, mice that received the high-frequency protocol showed a relatively greater degree of insulitis than the control mice.


Virtual optimization of nasal insulin therapy predicts immunization frequency to be crucial for diabetes protection.

Fousteri G, Chan JR, Zheng Y, Whiting C, Dave A, Bresson D, Croft M, von Herrath M - Diabetes (2010)

Nasal B:9–23 administration started at 4 weeks of age upon low-frequency immunization decreases insulitis and increases Treg in the islets. A–C: Reduced insulitis in low-frequency B:9-23 nasal immunized NOD. A: Pancreatic sections from B:9-23 nasally treated and control (PBS) mice were scored as described in research design and methods. The percentages represent the number of islets of a given score over the total number of islets examined. Scoring was performed on histological sections derived from 12-week-old nondiabetic mice (n = 4 mice per group). B: Pancreatic histology. Pancreatic sections (6 μm thick) were stained for insulin (blue) and CD8 (red) (C) or CD4 (red) (D) and analyzed at ×10 magnification. All histological sections shown were derived from 12-week-old nondiabetic mice. D–F: Increased Treg numbers in the islets of B:9-23–immunized mice. D: In silico–predicted changes in Treg levels within the pancreatic islet. The green box indicates the suggested-by-the-model-age window at which the laboratory measurements should be conducted. E: Actual Treg frequency as determined by measuring Foxp3+ cells per CD4+ cells per islet according to degree of insulitis. Same histological sections as in B and C were analyzed after immunofluorescent staining (IFS) for CD4, insulin, and Foxp3. Data are means ± SE. F: Representative microphotograph from each group of mice with insulitis degree 4 is shown at ×20 magnification (IFS for CD4 in blue, insulin in red, and Foxp3 in green). *P < 0.05 between PBS and high-frequency, high-frequency versus low-frequency immunization. **P < 0.01 between PBS and low-frequency immunization. (A high-quality digital representation of this figure is available in the online issue.)
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2992777&req=5

Figure 4: Nasal B:9–23 administration started at 4 weeks of age upon low-frequency immunization decreases insulitis and increases Treg in the islets. A–C: Reduced insulitis in low-frequency B:9-23 nasal immunized NOD. A: Pancreatic sections from B:9-23 nasally treated and control (PBS) mice were scored as described in research design and methods. The percentages represent the number of islets of a given score over the total number of islets examined. Scoring was performed on histological sections derived from 12-week-old nondiabetic mice (n = 4 mice per group). B: Pancreatic histology. Pancreatic sections (6 μm thick) were stained for insulin (blue) and CD8 (red) (C) or CD4 (red) (D) and analyzed at ×10 magnification. All histological sections shown were derived from 12-week-old nondiabetic mice. D–F: Increased Treg numbers in the islets of B:9-23–immunized mice. D: In silico–predicted changes in Treg levels within the pancreatic islet. The green box indicates the suggested-by-the-model-age window at which the laboratory measurements should be conducted. E: Actual Treg frequency as determined by measuring Foxp3+ cells per CD4+ cells per islet according to degree of insulitis. Same histological sections as in B and C were analyzed after immunofluorescent staining (IFS) for CD4, insulin, and Foxp3. Data are means ± SE. F: Representative microphotograph from each group of mice with insulitis degree 4 is shown at ×20 magnification (IFS for CD4 in blue, insulin in red, and Foxp3 in green). *P < 0.05 between PBS and high-frequency, high-frequency versus low-frequency immunization. **P < 0.01 between PBS and low-frequency immunization. (A high-quality digital representation of this figure is available in the online issue.)
Mentions: To determine the extent of islet infiltration following nasal B:9-23 immunization, pancreata from nondiabetic mice of all treatment groups were histologically examined at 12 weeks of age. As shown in Fig. 4A–C, the majority of islets in animals treated with the low-frequency protocol was minimally inflamed or had mild peri-insulitis. In contrast, most of the islets of normoglycemic mice that received the high-frequency immunization protocol had much greater CD4+ and CD8+ T-cell infiltrates. Furthermore, when scoring of the islets was undertaken, mice that received the high-frequency protocol showed a relatively greater degree of insulitis than the control mice.

Bottom Line: The experimental aim was to evaluate the impact of dose, frequency of administration, and age at treatment on Treg induction and optimal therapeutic outcome.Here, the advantage of applying computer modeling in optimizing the therapeutic efficacy of nasal insulin immunotherapy was confirmed.In silico modeling was able to streamline the experimental design and to identify the particular time frame at which biomarkers associated with protection in live NODs were induced.

View Article: PubMed Central - PubMed

Affiliation: Diabetes Center, La Jolla Institute for Allergy and Immunology, La Jolla, California, USA.

ABSTRACT

Objective: Development of antigen-specific strategies to treat or prevent type 1 diabetes has been slow and difficult because of the lack of experimental tools and defined biomarkers that account for the underlying therapeutic mechanisms.

Research design and methods: The type 1 diabetes PhysioLab platform, a large-scale mathematical model of disease pathogenesis in the nonobese diabetic (NOD) mouse, was used to investigate the possible mechanisms underlying the efficacy of nasal insulin B:9-23 peptide therapy. The experimental aim was to evaluate the impact of dose, frequency of administration, and age at treatment on Treg induction and optimal therapeutic outcome.

Results: In virtual NOD mice, treatment efficacy was predicted to depend primarily on the immunization frequency and stage of the disease and to a lesser extent on the dose. Whereas low-frequency immunization protected from diabetes atrributed to Treg and interleukin (IL)-10 induction in the pancreas 1-2 weeks after treatment, high-frequency immunization failed. These predictions were confirmed with wet-lab approaches, where only low-frequency immunization started at an early disease stage in the NOD mouse resulted in significant protection from diabetes by inducing IL-10 and Treg.

Conclusions: Here, the advantage of applying computer modeling in optimizing the therapeutic efficacy of nasal insulin immunotherapy was confirmed. In silico modeling was able to streamline the experimental design and to identify the particular time frame at which biomarkers associated with protection in live NODs were induced. These results support the development and application of humanized platforms for the design of clinical trials (i.e., for the ongoing nasal insulin prevention studies).

Show MeSH
Related in: MedlinePlus