Autoantigen-specific regulatory T cells induced in patients with type 1 diabetes mellitus by insulin B-chain immunotherapy
Tihamer Orban, Klara Farkas, Heyam Jalahej, Janos Kis, Andras Treszl, Ben Falk, Helena Reijonen, Joseph Wolfsdorf, Alyne Ricker, Jeffrey B Matthews, Nadio Tchao, Peter Sayre, Pete Bianchine, Tihamer Orban, Klara Farkas, Heyam Jalahej, Janos Kis, Andras Treszl, Ben Falk, Helena Reijonen, Joseph Wolfsdorf, Alyne Ricker, Jeffrey B Matthews, Nadio Tchao, Peter Sayre, Pete Bianchine
Abstract
There is a growing body of evidence to suggest that the autoimmunity observed in type 1 diabetes mellitus (T1DM) is the result of an imbalance between autoaggressive and regulatory cell subsets. Therapeutics that supplement or enhance the existing regulatory subset are therefore a much sought after goal in this indication. Here, we report the results of a double blind, placebo controlled, phase I clinical trial of a novel antigen-specific therapeutic in 12 subjects with recently diagnosed T1DM. Our primary objective was to test its safety. The study drug, human insulin B-chain in incomplete Freund's adjuvant (IFA) was administered as a single intramuscular injection, with subjects followed for 2 years. All subjects completed therapy and all follow-up visits. The therapy was generally safe and well-tolerated. Mixed meal stimulated C-peptide responses, measured every 6 months, showed no statistical differences between arms. All patients vaccinated with the autoantigen, but none who received placebo, developed robust insulin-specific humoral and T cell responses. Up to two years following the single injection, in peripheral blood from subjects in the experimental arm, but not the control arm, insulin B-chain-specific CD4+ T cells could be isolated and cloned that showed phenotypic and functional characteristics of regulatory T cells. The induction of a lasting, robust immune response generating autoantigen-specific regulatory T cells provides strong justification for further testing of this therapy in type 1 diabetes. (clinicaltrials.gov identifier NCT00057499).
Figures
![Figure 1](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/2860016/bin/nihms157062f1.jpg)
Figure 2
Insulin autoantibodies (IAA) by group…
Figure 2
Insulin autoantibodies (IAA) by group over time. IAA titers were significantly increased in…
Figure 3
T-cell assay stimulation indices (SI)…
Figure 3
T-cell assay stimulation indices (SI) in treatment and placebo groups in response to…
Figure 4
Ratio of the TGF-β1 concentrations…
Figure 4
Ratio of the TGF-β1 concentrations of the B-chain stimulated/unstimulated cells in the 7-day…
Figure 5
Suppression assay of two CFSE-B-chain…
Figure 5
Suppression assay of two CFSE-B-chain single cell clones with different cell-to-cell ratios of…
- Effects of high-dose oral insulin on immune responses in children at high risk for type 1 diabetes: the Pre-POINT randomized clinical trial.Bonifacio E, Ziegler AG, Klingensmith G, Schober E, Bingley PJ, Rottenkolber M, Theil A, Eugster A, Puff R, Peplow C, Buettner F, Lange K, Hasford J, Achenbach P; Pre-POINT Study Group. Bonifacio E, et al. JAMA. 2015 Apr 21;313(15):1541-9. doi: 10.1001/jama.2015.2928. JAMA. 2015. PMID: 25898052 Clinical Trial.
- Upregulating CD4+CD25+FOXP3+ regulatory T cells in pancreatic lymph nodes in diabetic NOD mice by adjuvant immunotherapy.Tian B, Hao J, Zhang Y, Tian L, Yi H, O'Brien TD, Sutherland DE, Hering BJ, Guo Z. Tian B, et al. Transplantation. 2009 Jan 27;87(2):198-206. doi: 10.1097/TP.0b013e3181933261. Transplantation. 2009. PMID: 19155973
- Proinsulin-specific T regulatory cells may control immune responses in type 1 diabetes: implications for adoptive therapy.Gliwiński M, Iwaszkiewicz-Grześ D, Wołoszyn-Durkiewicz A, Tarnowska M, Żalińska M, Hennig M, Zielińska H, Dukat-Mazurek A, Zielkowska-Dębska J, Zieliński M, Jaźwińska-Curyłło A, Owczuk R, Jarosz-Chobot P, Bossowski A, Szadkowska A, Młynarski W, Marek-Trzonkowska N, Moszkowska G, Siebert J, Myśliwiec M, Trzonkowski P. Gliwiński M, et al. BMJ Open Diabetes Res Care. 2020 Feb;8(1):e000873. doi: 10.1136/bmjdrc-2019-000873. BMJ Open Diabetes Res Care. 2020. PMID: 32098895 Free PMC article.
- Oral insulin immunotherapy in children at risk for type 1 diabetes in a randomised controlled trial.Assfalg R, Knoop J, Hoffman KL, Pfirrmann M, Zapardiel-Gonzalo JM, Hofelich A, Eugster A, Weigelt M, Matzke C, Reinhardt J, Fuchs Y, Bunk M, Weiss A, Hippich M, Halfter K, Hauck SM, Hasford J, Petrosino JF, Achenbach P, Bonifacio E, Ziegler AG. Assfalg R, et al. Diabetologia. 2021 May;64(5):1079-1092. doi: 10.1007/s00125-020-05376-1. Epub 2021 Jan 30. Diabetologia. 2021. PMID: 33515070 Free PMC article. Clinical Trial.
- Low-dose IL-2 in children with recently diagnosed type 1 diabetes: a Phase I/II randomised, double-blind, placebo-controlled, dose-finding study.Rosenzwajg M, Salet R, Lorenzon R, Tchitchek N, Roux A, Bernard C, Carel JC, Storey C, Polak M, Beltrand J, Amouyal C, Hartemann A, Corbeau P, Vicaut E, Bibal C, Bougnères P, Tran TA, Klatzmann D. Rosenzwajg M, et al. Diabetologia. 2020 Sep;63(9):1808-1821. doi: 10.1007/s00125-020-05200-w. Epub 2020 Jul 1. Diabetologia. 2020. PMID: 32607749 Clinical Trial.
- Delivery route considerations for designing antigen-specific biomaterial strategies to combat autoimmunity.Ackun-Farmmer MA, Jewell CM. Ackun-Farmmer MA, et al. Adv Nanobiomed Res. 2023 Mar;3(3):2200135. doi: 10.1002/anbr.202200135. Epub 2023 Jan 29. Adv Nanobiomed Res. 2023. PMID: 36938103
- Mechanisms and therapeutic strategies of immune checkpoint molecules and regulators in type 1 diabetes.Ding JT, Yang KP, Lin KL, Cao YK, Zou F. Ding JT, et al. Front Endocrinol (Lausanne). 2023 Jan 10;13:1090842. doi: 10.3389/fendo.2022.1090842. eCollection 2022. Front Endocrinol (Lausanne). 2023. PMID: 36704045 Free PMC article. Review.
- Safety of the use of gold nanoparticles conjugated with proinsulin peptide and administered by hollow microneedles as an immunotherapy in type 1 diabetes.Tatovic D, McAteer MA, Barry J, Barrientos A, Rodríguez Terradillos K, Perera I, Kochba E, Levin Y, Dul M, Coulman SA, Birchall JC, von Ruhland C, Howell A, Stenson R, Alhadj Ali M, Luzio SD, Dunseath G, Cheung WY, Holland G, May K, Ingram JR, Chowdhury MMU, Wong FS, Casas R, Dayan C, Ludvigsson J. Tatovic D, et al. Immunother Adv. 2022 Jan 27;2(1):ltac002. doi: 10.1093/immadv/ltac002. eCollection 2022. Immunother Adv. 2022. PMID: 35919496 Free PMC article.
- Potential Therapeutic Application of Regulatory T Cells in Diabetes Mellitus Type 1.Ben-Skowronek I, Sieniawska J, Pach E, Wrobel W, Skowronek A, Tomczyk Z, Rosolowska I. Ben-Skowronek I, et al. Int J Mol Sci. 2021 Dec 30;23(1):390. doi: 10.3390/ijms23010390. Int J Mol Sci. 2021. PMID: 35008819 Free PMC article. Review.
- Biologia Futura: Emerging antigen-specific therapies for autoimmune diseases.Sármay G. Sármay G. Biol Futur. 2021 Mar;72(1):15-24. doi: 10.1007/s42977-021-00074-4. Epub 2021 Feb 4. Biol Futur. 2021. PMID: 34554499 Review.
- Clinical Trial, Phase I
- Randomized Controlled Trial
- Research Support, N.I.H., Extramural
- Adolescent
- Adult
- Autoantigens / administration & dosage*
- Autoantigens / therapeutic use
- Autoimmunity / drug effects
- Cell Proliferation
- Clone Cells / immunology
- Diabetes Mellitus, Type 1 / immunology
- Diabetes Mellitus, Type 1 / therapy
- Double-Blind Method
- Humans
- Immunotherapy
- Insulin / administration & dosage*
- Insulin / immunology
- Insulin / therapeutic use
- T-Lymphocytes, Regulatory / drug effects
- T-Lymphocytes, Regulatory / immunology*
- Treatment Outcome
- Vaccination
- Young Adult
- Autoantigens
- Insulin
- ClinicalTrials.gov/NCT00057499
- Full Text Sources
- Other Literature Sources
- Medical
- Research Materials
NCBI Literature Resources
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![Figure 2](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/2860016/bin/nihms157062f2.jpg)
Figure 3
T-cell assay stimulation indices (SI)…
Figure 3
T-cell assay stimulation indices (SI) in treatment and placebo groups in response to…
Figure 4
Ratio of the TGF-β1 concentrations…
Figure 4
Ratio of the TGF-β1 concentrations of the B-chain stimulated/unstimulated cells in the 7-day…
Figure 5
Suppression assay of two CFSE-B-chain…
Figure 5
Suppression assay of two CFSE-B-chain single cell clones with different cell-to-cell ratios of…
- Effects of high-dose oral insulin on immune responses in children at high risk for type 1 diabetes: the Pre-POINT randomized clinical trial.Bonifacio E, Ziegler AG, Klingensmith G, Schober E, Bingley PJ, Rottenkolber M, Theil A, Eugster A, Puff R, Peplow C, Buettner F, Lange K, Hasford J, Achenbach P; Pre-POINT Study Group. Bonifacio E, et al. JAMA. 2015 Apr 21;313(15):1541-9. doi: 10.1001/jama.2015.2928. JAMA. 2015. PMID: 25898052 Clinical Trial.
- Upregulating CD4+CD25+FOXP3+ regulatory T cells in pancreatic lymph nodes in diabetic NOD mice by adjuvant immunotherapy.Tian B, Hao J, Zhang Y, Tian L, Yi H, O'Brien TD, Sutherland DE, Hering BJ, Guo Z. Tian B, et al. Transplantation. 2009 Jan 27;87(2):198-206. doi: 10.1097/TP.0b013e3181933261. Transplantation. 2009. PMID: 19155973
- Proinsulin-specific T regulatory cells may control immune responses in type 1 diabetes: implications for adoptive therapy.Gliwiński M, Iwaszkiewicz-Grześ D, Wołoszyn-Durkiewicz A, Tarnowska M, Żalińska M, Hennig M, Zielińska H, Dukat-Mazurek A, Zielkowska-Dębska J, Zieliński M, Jaźwińska-Curyłło A, Owczuk R, Jarosz-Chobot P, Bossowski A, Szadkowska A, Młynarski W, Marek-Trzonkowska N, Moszkowska G, Siebert J, Myśliwiec M, Trzonkowski P. Gliwiński M, et al. BMJ Open Diabetes Res Care. 2020 Feb;8(1):e000873. doi: 10.1136/bmjdrc-2019-000873. BMJ Open Diabetes Res Care. 2020. PMID: 32098895 Free PMC article.
- Oral insulin immunotherapy in children at risk for type 1 diabetes in a randomised controlled trial.Assfalg R, Knoop J, Hoffman KL, Pfirrmann M, Zapardiel-Gonzalo JM, Hofelich A, Eugster A, Weigelt M, Matzke C, Reinhardt J, Fuchs Y, Bunk M, Weiss A, Hippich M, Halfter K, Hauck SM, Hasford J, Petrosino JF, Achenbach P, Bonifacio E, Ziegler AG. Assfalg R, et al. Diabetologia. 2021 May;64(5):1079-1092. doi: 10.1007/s00125-020-05376-1. Epub 2021 Jan 30. Diabetologia. 2021. PMID: 33515070 Free PMC article. Clinical Trial.
- Low-dose IL-2 in children with recently diagnosed type 1 diabetes: a Phase I/II randomised, double-blind, placebo-controlled, dose-finding study.Rosenzwajg M, Salet R, Lorenzon R, Tchitchek N, Roux A, Bernard C, Carel JC, Storey C, Polak M, Beltrand J, Amouyal C, Hartemann A, Corbeau P, Vicaut E, Bibal C, Bougnères P, Tran TA, Klatzmann D. Rosenzwajg M, et al. Diabetologia. 2020 Sep;63(9):1808-1821. doi: 10.1007/s00125-020-05200-w. Epub 2020 Jul 1. Diabetologia. 2020. PMID: 32607749 Clinical Trial.
- Delivery route considerations for designing antigen-specific biomaterial strategies to combat autoimmunity.Ackun-Farmmer MA, Jewell CM. Ackun-Farmmer MA, et al. Adv Nanobiomed Res. 2023 Mar;3(3):2200135. doi: 10.1002/anbr.202200135. Epub 2023 Jan 29. Adv Nanobiomed Res. 2023. PMID: 36938103
- Mechanisms and therapeutic strategies of immune checkpoint molecules and regulators in type 1 diabetes.Ding JT, Yang KP, Lin KL, Cao YK, Zou F. Ding JT, et al. Front Endocrinol (Lausanne). 2023 Jan 10;13:1090842. doi: 10.3389/fendo.2022.1090842. eCollection 2022. Front Endocrinol (Lausanne). 2023. PMID: 36704045 Free PMC article. Review.
- Safety of the use of gold nanoparticles conjugated with proinsulin peptide and administered by hollow microneedles as an immunotherapy in type 1 diabetes.Tatovic D, McAteer MA, Barry J, Barrientos A, Rodríguez Terradillos K, Perera I, Kochba E, Levin Y, Dul M, Coulman SA, Birchall JC, von Ruhland C, Howell A, Stenson R, Alhadj Ali M, Luzio SD, Dunseath G, Cheung WY, Holland G, May K, Ingram JR, Chowdhury MMU, Wong FS, Casas R, Dayan C, Ludvigsson J. Tatovic D, et al. Immunother Adv. 2022 Jan 27;2(1):ltac002. doi: 10.1093/immadv/ltac002. eCollection 2022. Immunother Adv. 2022. PMID: 35919496 Free PMC article.
- Potential Therapeutic Application of Regulatory T Cells in Diabetes Mellitus Type 1.Ben-Skowronek I, Sieniawska J, Pach E, Wrobel W, Skowronek A, Tomczyk Z, Rosolowska I. Ben-Skowronek I, et al. Int J Mol Sci. 2021 Dec 30;23(1):390. doi: 10.3390/ijms23010390. Int J Mol Sci. 2021. PMID: 35008819 Free PMC article. Review.
- Biologia Futura: Emerging antigen-specific therapies for autoimmune diseases.Sármay G. Sármay G. Biol Futur. 2021 Mar;72(1):15-24. doi: 10.1007/s42977-021-00074-4. Epub 2021 Feb 4. Biol Futur. 2021. PMID: 34554499 Review.
- Clinical Trial, Phase I
- Randomized Controlled Trial
- Research Support, N.I.H., Extramural
- Adolescent
- Adult
- Autoantigens / administration & dosage*
- Autoantigens / therapeutic use
- Autoimmunity / drug effects
- Cell Proliferation
- Clone Cells / immunology
- Diabetes Mellitus, Type 1 / immunology
- Diabetes Mellitus, Type 1 / therapy
- Double-Blind Method
- Humans
- Immunotherapy
- Insulin / administration & dosage*
- Insulin / immunology
- Insulin / therapeutic use
- T-Lymphocytes, Regulatory / drug effects
- T-Lymphocytes, Regulatory / immunology*
- Treatment Outcome
- Vaccination
- Young Adult
- Autoantigens
- Insulin
- ClinicalTrials.gov/NCT00057499
- Full Text Sources
- Other Literature Sources
- Medical
- Research Materials
![Figure 3](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/2860016/bin/nihms157062f3.jpg)
![Figure 4](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/2860016/bin/nihms157062f4.jpg)
Figure 5
Suppression assay of two CFSE-B-chain…
Figure 5
Suppression assay of two CFSE-B-chain single cell clones with different cell-to-cell ratios of…
![Figure 5](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/2860016/bin/nihms157062f5.jpg)
Source: PubMed