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In vivo relationship between bound and free insulin in patients with diabetes having anti-insulin antibodies

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Abstract

Objective

The Scatchard plot of anti-insulin antibodies is curvilinear, indicating heterogeneity in binding sites. However, the relationship between bound insulin (B) and free insulin (F) in patients with anti-insulin antibodies has not yet been elucidated. This study aimed to determine this relationship.

Methods

We studied two insulin-treated patients with diabetes who had high titers of anti-insulin antibodies. The B and F levels were measured using daily blood samples. Assuming that the law of mass action is applicable to the reactions between insulin and anti-insulin antibody forms, we plotted the bound-to-free ratio (B/F) vs. B using patient data. We also performed an equilibrium binding assay in vitro.

Results

Some of the B/F vs. B plots of the daily variation showed an approximately linear relationship, while the Scatchard plots of in vitro data became curvilinear.

Conclusion

Our study suggests that the one-site (high-affinity site) of anti-insulin antibodies accounts, for the most part, for insulin pharmacokinetics within physiological insulin concentrations.

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Data availability

The deidentified participant data will be shared on a request basis. Please contact Hiroyuki Asaka (daitoku@muse.ocn.ne.jp). But the data used in this study has been restricted by the Kanazawa University IRB.

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Acknowledgements

We thank the patients who participated in this study.

Funding

None.

Author information

Authors and Affiliations

  1. Division of Endocrine and Diabetes, Department of Cardiovascular and Internal Medicine, Kanazawa University Graduate School of Medicine, 13-1 Takara-machi, Kanazawa, Ishikawa, 920-8641, Japan

    Hiroyuki Asaka, Mitsuhiro Kometani & Takashi Yoneda

  2. Daitoku Internal Medicine and Diabetes Clinic, 2-570 Uneda-higashi, Kanazawa, Ishikawa, 920-0344, Japan

    Hiroyuki Asaka

  3. Institute of Liberal Arts and Science, Kanazawa University, Kanazawa, 920-1192, Japan

    Shigehiro Karashima

  4. Center for Clinical Research, Toyama University Hospital, Sugitani 2630, Toyama, 930-0194, Japan

    Daisuke Chujo

  5. Internal Medicine, Houju Memorial Hospital, 11-71 Midorigaoka, Nomi, Ishikawa, 923-1226, Japan

    Mikiya Usukura

  6. Center for Clinical Genomics, Kanazawa Medical University Hospital, 1-1 Daigaku, Uchinada, Ishikawa, 920-0293, Japan

    Kunimasa Yagi

  7. Department of Health Promotion and Medicine of the Future, Graduate School of Medicine, Kanazawa University, Kanazawa, 920-8541, Japan

    Ko Aiga & Takashi Yoneda

  8. Faculty of Transdisciplinary Sciences, Institute of Transdisciplinary Sciences, Kanazawa University, Kanazawa, 920-1192, Japan

    Takashi Yoneda

Authors
  1. Hiroyuki Asaka
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  2. Shigehiro Karashima
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  3. Daisuke Chujo
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  4. Mitsuhiro Kometani
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  5. Mikiya Usukura
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  6. Kunimasa Yagi
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  7. Ko Aiga
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  8. Takashi Yoneda
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Contributions

HA conceived and designed the study, analyzed the data, and drafted the manuscript for publication. SK designed the study and reviewed the manuscript. DC supervised the study, contributed to discussion, and reviewed and edited the manuscript. MK, MU, KY, KA, and TY reviewed the manuscript. All authors edited, reviewed, and approved the final version of the manuscript. HA is the guarantor of this work, and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Corresponding author

Correspondence to Hiroyuki Asaka.

Ethics declarations

Conflict of interest

None of the authors have any potential conflicts of interest associated with this research.

Human and animal rights

All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and/or with the Helsinki Declaration of 1964 and later versions. This study was approved by the ethics committee of Kanazawa University (approval No. 1917-1/ September 16, 2015, approval No. 1917-2/February 21, 2017).

Informed consent

Written informed consent was obtained from the patients.

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Supplementary Information

Below is the link to the electronic supplementary material.

13340_2023_641_MOESM1_ESM.pdf

Supplementary file1 The results of the patient with type 1 diabetes mellitus who had moderate anti-insulin antibodies (Patient 3). A: Fasting data. B: Daily variation in total insulin, free insulin, and plasma glucose. C: Scatchard plot (B/F vs. B plot) and Ka and Bmax by one-site model in vivo. D: Scatchard plot using 125I-labeled porcine insulin and Ka and Bmax by the two-site model in vitro. The patient was treated with continuous subcutaneous insulin infusion with insulin lispro (4.0, 6.9, and 6.7 units before breakfast, lunch, and dinner, respectively) and total daily basal insulin (15.6 units/day). T-IRI, total immunoreactive insulin (pmol/L); F-IRI, free immunoreactive insulin (pmol/L); PG, plasma glucose (mg/dL); B, bound insulin (10−8 M); F, free insulin; B/F, bound/free ratio; Ka, affinity constant (1/10−8 M); Bmax, binding capacity (10−8 M) (PDF 68 KB)

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Asaka, H., Karashima, S., Chujo, D. et al. In vivo relationship between bound and free insulin in patients with diabetes having anti-insulin antibodies. Diabetol Int 14, 427–433 (2023). https://doi.org/10.1007/s13340-023-00641-1

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  • DOI: https://doi.org/10.1007/s13340-023-00641-1

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