Genetic and structural basis of the human anti-α-galactosyl antibody response

Langley, David B.; Schofield, Peter; Névoltris, Damien; Jackson, Jennifer; Jackson, Katherine; Peters, Tim J; Burk, Melanie; Matthews, J.M.; Basten, Antony; Goodnow, Christopher C.; Nunen, Sheryl van; Reed, Joanne H.; Christ, Daniel

doi:10.1073/pnas.2123212119

Cited by 7 publications

(5 citation statements)

References 71 publications

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“…We observed a significant increase in the production of the heavy and light chains at 1 dpi following bacterial bioreactor growth, a consistent increase in the heavy chain at 3 dpi, however, by 7 dpi, there was no significant differences in trastuzumab production regardless of the bacterial growth condition (Figure 3e). We confirmed trastuzumab production at 3 and 7 dpi via western blot for both agroinfiltration methods (Figure 3f; Figure S2) and further validated the increase in trastuzumab production at 3 dpi following bioreactor infiltration using the Blitz system (the industry‐standard method for quantifying target protein production; Langley et al ., 2022; Lua et al ., 2015; Figure 3g).…”

Section: Resultsmentioning

confidence: 99%

Bacterial growth‐mediated systems remodelling of Nicotiana benthamiana defines unique signatures of target protein production in molecular pharming

Prudhomme,

Pastora,

Thomson

et al. 2024

Plant Biotechnology Journal

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SummaryThe need for therapeutics to treat a plethora of medical conditions and diseases is on the rise and the demand for alternative approaches to mammalian‐based production systems is increasing. Plant‐based strategies provide a safe and effective alternative to produce biological drugs but have yet to enter mainstream manufacturing at a competitive level. Limitations associated with batch consistency and target protein production levels are present; however, strategies to overcome these challenges are underway. In this study, we apply state‐of‐the‐art mass spectrometry‐based proteomics to define proteome remodelling of the plant following agroinfiltration with bacteria grown under shake flask or bioreactor conditions. We observed distinct signatures of bacterial protein production corresponding to the different growth conditions that directly influence the plant defence responses and target protein production on a temporal axis. Our integration of proteomic profiling with small molecule detection and quantification reveals the fluctuation of secondary metabolite production over time to provide new insight into the complexities of dual system modulation in molecular pharming. Our findings suggest that bioreactor bacterial growth may promote evasion of early plant defence responses towards Agrobacterium tumefaciens (updated nomenclature to Rhizobium radiobacter). Furthermore, we uncover and explore specific targets for genetic manipulation to suppress host defences and increase recombinant protein production in molecular pharming.

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Section: Resultsmentioning

confidence: 99%

Bacterial growth‐mediated systems remodelling of Nicotiana benthamiana defines unique signatures of target protein production in molecular pharming

Prudhomme,

Pastora,

Thomson

et al. 2024

Plant Biotechnology Journal

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“…Due to gut microbiota expressing α-Gal, anti-Gal antibodies are one of the most abundant types of antibodies in humans ( 20 , 21 ), constituting 0.2 - 1.0% of total immunoglobulins ( 22 , 23 ). Anti-Gal antibodies are characterized by a preferential use of Ig variable region V3 family genes in the heavy chain ( 24 – 26 ), where a germline encoded tryptophan in position 33 of the complementarity determining region is essential for the binding to α-Gal ( 26 ). The formation of anti-Gal antibodies begins in the first few months of life and the IgM isotype rises faster and steeper compared to IgG and IgA until 2 years of age ( 27 ).…”

Section: The α-Gal Epitope – a Strong Immunogenmentioning

confidence: 99%

“…However, allergen-specific IgG+ MBC have been clonally linked to IgE+ plasma blasts ( 92 ), and IgG+ MBC that differentiate into IgE+ plasma cells have been shown to have a specific surface phenotype, expressing CD23 and the IL-4R ( 93 – 95 ). In AGS patients, no α-Gal-specific IgE+ B cells were found in circulation, but α-Gal-specific B cells had similar usage of B cell receptor heavy chain V genes as healthy individuals, that were mainly of the IgM isotype ( 26 ). However, there is evidence that human IgE+ B cells mainly seem to have differentiated from IgG1+ B cells through sequential class-switching ( 92 , 96 , 97 ).…”

Section: Mechanisms Of Sensitization To α-Galmentioning

confidence: 99%

The α-Gal epitope - the cause of a global allergic disease

Perusko,

Grundström,

Eldh

et al. 2024

Front. Immunol.

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The galactose-α-1,3-galactose (α-Gal) epitope is the cause of a global allergic disease, the α-Gal syndrome (AGS). It is a severe form of allergy to food and products of mammalian origin where IgE against the mammalian carbohydrate, α-Gal, is the cause of the allergic reactions. Allergic reactions triggered by parenterally administered α-Gal sources appear immediately, but those triggered via the oral route appear with a latency of several hours. The α-Gal epitope is highly immunogenic to humans, apes and old-world monkeys, all of which produce anti-α-Gal antibodies of the IgM, IgA and IgG subclasses. Strong evidence suggests that in susceptible individuals, class switch to IgE occurs after several tick bites. In this review, we discuss the strong immunogenic role of the α-Gal epitope and its structural resemblance to the blood type B antigen. We emphasize the broad abundance of α-Gal in different foods and pharmaceuticals and the allergenicity of various α-Gal containing molecules. We give an overview of the association of tick bites with the development of AGS and describe innate and adaptive immune response to tick saliva that possibly leads to sensitization to α-Gal. We further discuss a currently favored hypothesis explaining the mechanisms of the delayed effector phase of the allergic reaction to α-Gal. We highlight AGS from a clinical point of view. We review the different clinical manifestations of the disease and the prevalence of sensitization to α-Gal and AGS. The usefulness of various diagnostic tests is discussed. Finally, we provide different aspects of the management of AGS. With climate change and global warming, the tick density is increasing, and their geographic range is expanding. Thus, more people will be affected by AGS which requires more knowledge of the disease.

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“…A crystallization study with the monoclonal anti-Gal antibody M86 immunocomplexed with the Galα1-3Gal portion of the α-gal epitope demonstrated a groove in the binding site of this antibody, in which the Galα1-3Gal disaccharide binds via hydrogen bonds to the antibody ( Langley et al, 2022 ). In view of this study, it is possible that the groove shape in the binding site of pure anti-Gal antibody clones differs from that of anti-Gal/B in that in the latter, the binding is not affected by the fucose-linked α1-2 to the penultimate galactose, as schematically shown in Figure 1 ( Galili et al, 1987b ).…”

Section: Anti-gal Immune Response In Humansmentioning

confidence: 99%

Antibody production and tolerance to the α-gal epitope as models for understanding and preventing the immune response to incompatible ABO carbohydrate antigens and for α-gal therapies

Galili

2023

Front. Mol. Biosci.

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This review describes the significance of the α-gal epitope (Galα-3Galβ1-4GlcNAc-R) as the core of human blood-group A and B antigens (A and B antigens), determines in mouse models the principles underlying the immune response to these antigens, and suggests future strategies for the induction of immune tolerance to incompatible A and B antigens in human allografts. Carbohydrate antigens, such as ABO antigens and the α-gal epitope, differ from protein antigens in that they do not interact with T cells, but B cells interacting with them require T-cell help for their activation. The α-gal epitope is the core of both A and B antigens and is the ligand of the natural anti-Gal antibody, which is abundant in all humans. In A and O individuals, anti-Gal clones (called anti-Gal/B) comprise >85% of the so-called anti-B activity and bind to the B antigen in facets that do not include fucose-linked α1–2 to the core α-gal. As many as 1% of B cells are anti-Gal B cells. Activation of quiescent anti-Gal B cells upon exposure to α-gal epitopes on xenografts and some protozoa can increase the titer of anti-Gal by 100-fold. α1,3-Galactosyltransferase knockout (GT-KO) mice lack α-gal epitopes and can produce anti-Gal. These mice simulate human recipients of ABO-incompatible human allografts. Exposure for 2–4 weeks of naïve and memory mouse anti-Gal B cells to α-gal epitopes in the heterotopically grafted wild-type (WT) mouse heart results in the elimination of these cells and immune tolerance to this epitope. Shorter exposures of 7 days of anti-Gal B cells to α-gal epitopes in the WT heart result in the production of accommodating anti-Gal antibodies that bind to α-gal epitopes but do not lyse cells or reject the graft. Tolerance to α-gal epitopes due to the elimination of naïve and memory anti-Gal B cells can be further induced by 2 weeks in vivo exposure to WT lymphocytes or autologous lymphocytes engineered to present α-gal epitopes by transduction of the α1,3-galactosyltransferase gene. These mouse studies suggest that autologous human lymphocytes similarly engineered to present the A or B antigen may induce corresponding tolerance in recipients of ABO-incompatible allografts. The review further summarizes experimental works demonstrating the efficacy of α-gal therapies in amplifying anti-viral and anti-tumor immune-protection and regeneration of injured tissues.

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Genetic and structural basis of the human anti-α-galactosyl antibody response

Cited by 7 publications

References 71 publications

Bacterial growth‐mediated systems remodelling of Nicotiana benthamiana defines unique signatures of target protein production in molecular pharming

Bacterial growth‐mediated systems remodelling of Nicotiana benthamiana defines unique signatures of target protein production in molecular pharming

The α-Gal epitope - the cause of a global allergic disease

Antibody production and tolerance to the α-gal epitope as models for understanding and preventing the immune response to incompatible ABO carbohydrate antigens and for α-gal therapies

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