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Rapid in vitro assessment of the immunogenicity potential of engineered antibody therapeutics through detection of CD4⁺ T cell interleukin-2 secretion.

ABSTRACT: Therapeutic antibodies sometimes elicit anti-drug antibodies (ADAs) that can affect efficacy and safety. Engineered antibodies that contain artificial amino acid sequences are potentially highly immunogenic, but this is currently difficult to predict. Therefore, it is important to efficiently assess immunogenicity during the development of complex antibody-based formats. Here, we present an in vitro peripheral blood mononuclear cell-based assay that can be used to assess immunogenicity potential within 3 days. This method involves examining the frequency and function of interleukin (IL)-2-secreting CD4⁺ T cells induced by therapeutic antibodies. IL-2-secreting CD4⁺ T cells seem to be functionally relevant to the immunogenic potential due to their proliferative activity and the expression of several cytokines. The rates of the donors responding to low and high immunogenic proteins, mAb1, and keyhole limpet hemocyanin were 1.3% and 93.5%, respectively. Seven antibodies with known rates of immunogenicity (etanercept, emicizumab, abciximab, romosozumab, blosozumab, humanized anti-human A33 antibody, and bococizumab) induced responses in 1.9%, 3.8%, 6.4%, 10.0%, 29.2%, 43.8%, and 89.5% of donors, respectively. These data are comparable with ADA incidences in clinical settings. Our results show that this assay can contribute to the swift assessment and mechanistic understanding of the immunogenicity of therapeutic antibodies.

SUBMITTER: Arata Y

PROVIDER: S-EPMC10494738 | biostudies-literature | 2023 Jan-Dec

REPOSITORIES: biostudies-literature

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Rapid <i>in vitro</i> assessment of the immunogenicity potential of engineered antibody therapeutics through detection of CD4<sup>+</sup> T cell interleukin-2 secretion.

Arata Yoshiyuki Y Motoyama Shigeki S Yano Mariko M Ikuno Tatsuya T Ito Shunsuke S Matsushita Tomochika T Takeiri Akira A Nishito Yukari Y Yabuki Nami N Mizuno Hideaki H Sampei Zenjiro Z Mishima Masayuki M Honda Masaki M Kiyokawa Jumpei J Suzuki Hiromi H Chiba Shuichi S Tabo Mitsuyasu M Kubo Chiyomi C

mAbs 20230101 1

Therapeutic antibodies sometimes elicit anti-drug antibodies (ADAs) that can affect efficacy and safety. Engineered antibodies that contain artificial amino acid sequences are potentially highly immunogenic, but this is currently difficult to predict. Therefore, it is important to efficiently assess immunogenicity during the development of complex antibody-based formats. Here, we present an in vitro peripheral blood mononuclear cell-based assay that can be used to assess immunogenicity potential ...[more]

PMID: 37682072

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Project description:BackgroundThe immune system and the skeletal system have complex interactions in the bone marrow and even in the joints, which has promoted the development of the concept of osteoimmunology. Some evidence has indicated that T cells and B cells contribute to the balance between the resorption and formation of bone. However, there has been little discussion on the regulation of CD4+ T lymphocytes by cells involved in bone metabolism. Mesenchymal stem cells (MSCs), which exert core functions related to immunoregulation and osteogenic differentiation, are crucial cells linked to both bone metabolism and the immune system. Previous studies have shown that the immunoregulatory capacity of MSCs changes following differentiation. However, it is still unclear whether the osteogenic differentiation of MSCs affects the migration and differentiation of CD4+ T cells.MethodsMSCs were cultured in growth medium or osteogenic medium for 10 days and then cocultured with CD4+ T cells. CD4+ T cell migration and differentiation were detected by flow cytometry. Further, gene expression levels of specific cytokines were analyzed by quantitative real-time PCR and enzyme-linked immunosorbent assays. A Proteome Profiler Human XL Cytokine Array Kit was used to analyze supernatants collected from MSCs. Alizarin red S staining and Alkaline phosphatase assay were used to detect the osteogenic differentiation of MSCs.ResultsHere, we found that the migration of CD4+ T cells was elevated, and the capacity to induce the differentiation of regulatory T (Treg) cells was weakened during MSC osteogenic differentiation, while the differentiation of T helper 1 (Th1), T helper 2 (Th2) and T helper 17 (Th17) cells was not affected. Further studies revealed that interleukin (IL)-8 was significantly upregulated during MSC osteogenic differentiation. Both a neutralizing antibody and IL-8-specific siRNA significantly inhibited the migration of CD4+ T cells and promoted the differentiation of Treg cells. Finally, we found that the transcription factor c-Jun was involved in regulating the expression of IL-8 and affected the osteogenic differentiation of MSCs, thereby mediating the migration and differentiation of CD4+ T cells.ConclusionThis study demonstrated that MSC osteogenic differentiation promoted c-Jun-dependent secretion of IL-8 and mediated the migration and differentiation of CD4+ T cells. These results provide a further understanding of the crosstalk between bone and the immune system and reveal information about the relationship between osteogenesis and inflammation in the field of osteoimmunology.

Dataset Information

Rapid in vitro assessment of the immunogenicity potential of engineered antibody therapeutics through detection of CD4⁺ T cell interleukin-2 secretion.

Publications

Rapid <i>in vitro</i> assessment of the immunogenicity potential of engineered antibody therapeutics through detection of CD4<sup>+</sup> T cell interleukin-2 secretion.

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Dataset Information

Rapid in vitro assessment of the immunogenicity potential of engineered antibody therapeutics through detection of CD4+ T cell interleukin-2 secretion.

Publications

Rapid <i>in vitro</i> assessment of the immunogenicity potential of engineered antibody therapeutics through detection of CD4<sup>+</sup> T cell interleukin-2 secretion.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Rapid in vitro assessment of the immunogenicity potential of engineered antibody therapeutics through detection of CD4⁺ T cell interleukin-2 secretion.