Enhanced effect of the immunosuppressive soluble HLA-G2 homodimer by site-specific PEGylation
Abstract Human leukocyte antigen (HLA)-G is a nonclassical HLA class I molecule that has an immunosuppressive effect mediated by binding to immune inhibitory leukocyte immunoglobulin-like receptors (LILR) B1 and LILRB2. A conventional HLA-G isoform, HLA-G1, forms a heterotrimeric complex composed of...
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Nature Portfolio
2025-01-01
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| author | Chisato Yamada Kimiko Kuroki Naoyoshi Maeda Hiroshi Watanabe Ami Takahashi Katsumi Maenaka |
| author_facet | Chisato Yamada Kimiko Kuroki Naoyoshi Maeda Hiroshi Watanabe Ami Takahashi Katsumi Maenaka |
| author_sort | Chisato Yamada |
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| description | Abstract Human leukocyte antigen (HLA)-G is a nonclassical HLA class I molecule that has an immunosuppressive effect mediated by binding to immune inhibitory leukocyte immunoglobulin-like receptors (LILR) B1 and LILRB2. A conventional HLA-G isoform, HLA-G1, forms a heterotrimeric complex composed of a heavy chain (α1-α3 domains), β2-microglobulin (β2m) and a cognate peptide. One of the other isoforms, HLA-G2, lacks a α2 domain or β2m to form a nondisulfide-linked homodimer, and its ectodomain specifically binds to LILRB2 expressed in human monocytes, macrophages, and dendritic cells. The administration of the ectodomain of HLA-G2, designated the soluble HLA-G2 homodimer, showed significant immunosuppressive effects in mouse models of rheumatoid arthritis and systemic lupus erythematosus, presumably by binding to a mouse ortholog of LILRB2, paired immunoglobulin-like receptor B. However, the refolded soluble HLA-G2 homodimer used in these studies tends to aggregate and degrade; thus, its stability for clinical use has been a concern. In the present study, we improved the stability of the refolded soluble HLA-G2 homodimer via a site-directed PEGylation method. PEGylation at an original free cysteine residue, Cys42, resulted in increased lyophilization and thermal and serum stability. Furthermore, the PEGylated soluble HLA-G2 homodimer could better suppress atopic symptoms in mice than the non-PEGylated homodimer. These results suggest that PEGylated soluble HLA-G2 homodimers could be candidates for immunosuppressive biologics that specifically target LILRB2-positive myelomonocytic antigen-presenting cells. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-ff61e90416b84d3b82a0f67077d494cf2025-01-26T12:26:08ZengNature PortfolioScientific Reports2045-23222025-01-0115111410.1038/s41598-024-85072-xEnhanced effect of the immunosuppressive soluble HLA-G2 homodimer by site-specific PEGylationChisato Yamada0Kimiko Kuroki1Naoyoshi Maeda2Hiroshi Watanabe3Ami Takahashi4Katsumi Maenaka5Laboratory of Biomolecular Science, Faculty of Pharmaceutical Sciences, Hokkaido UniversityLaboratory of Biomolecular Science, Faculty of Pharmaceutical Sciences, Hokkaido UniversityCenter for Research and Education On Drug Discovery, Faculty of Pharmaceutical Sciences, Hokkaido UniversityLaboratory of Biomolecular Science, Faculty of Pharmaceutical Sciences, Hokkaido UniversityLaboratory of Biomolecular Science, Faculty of Pharmaceutical Sciences, Hokkaido UniversityLaboratory of Biomolecular Science, Faculty of Pharmaceutical Sciences, Hokkaido UniversityAbstract Human leukocyte antigen (HLA)-G is a nonclassical HLA class I molecule that has an immunosuppressive effect mediated by binding to immune inhibitory leukocyte immunoglobulin-like receptors (LILR) B1 and LILRB2. A conventional HLA-G isoform, HLA-G1, forms a heterotrimeric complex composed of a heavy chain (α1-α3 domains), β2-microglobulin (β2m) and a cognate peptide. One of the other isoforms, HLA-G2, lacks a α2 domain or β2m to form a nondisulfide-linked homodimer, and its ectodomain specifically binds to LILRB2 expressed in human monocytes, macrophages, and dendritic cells. The administration of the ectodomain of HLA-G2, designated the soluble HLA-G2 homodimer, showed significant immunosuppressive effects in mouse models of rheumatoid arthritis and systemic lupus erythematosus, presumably by binding to a mouse ortholog of LILRB2, paired immunoglobulin-like receptor B. However, the refolded soluble HLA-G2 homodimer used in these studies tends to aggregate and degrade; thus, its stability for clinical use has been a concern. In the present study, we improved the stability of the refolded soluble HLA-G2 homodimer via a site-directed PEGylation method. PEGylation at an original free cysteine residue, Cys42, resulted in increased lyophilization and thermal and serum stability. Furthermore, the PEGylated soluble HLA-G2 homodimer could better suppress atopic symptoms in mice than the non-PEGylated homodimer. These results suggest that PEGylated soluble HLA-G2 homodimers could be candidates for immunosuppressive biologics that specifically target LILRB2-positive myelomonocytic antigen-presenting cells.https://doi.org/10.1038/s41598-024-85072-xHLA-G2LILRPEGylationImmune checkpointImmunosuppressive |
| spellingShingle | Chisato Yamada Kimiko Kuroki Naoyoshi Maeda Hiroshi Watanabe Ami Takahashi Katsumi Maenaka Enhanced effect of the immunosuppressive soluble HLA-G2 homodimer by site-specific PEGylation Scientific Reports HLA-G2 LILR PEGylation Immune checkpoint Immunosuppressive |
| title | Enhanced effect of the immunosuppressive soluble HLA-G2 homodimer by site-specific PEGylation |
| title_full | Enhanced effect of the immunosuppressive soluble HLA-G2 homodimer by site-specific PEGylation |
| title_fullStr | Enhanced effect of the immunosuppressive soluble HLA-G2 homodimer by site-specific PEGylation |
| title_full_unstemmed | Enhanced effect of the immunosuppressive soluble HLA-G2 homodimer by site-specific PEGylation |
| title_short | Enhanced effect of the immunosuppressive soluble HLA-G2 homodimer by site-specific PEGylation |
| title_sort | enhanced effect of the immunosuppressive soluble hla g2 homodimer by site specific pegylation |
| topic | HLA-G2 LILR PEGylation Immune checkpoint Immunosuppressive |
| url | https://doi.org/10.1038/s41598-024-85072-x |
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