Enhancement of immunogenic properties of ovalbumin as a result of its chlorination

doi:10.1016/0020-711X(91)90280-Z

International Journal of Biochemistry

Volume 23, Issue 12, 1991, Pages 1393-1395

https://doi.org/10.1016/0020-711X(91)90280-Z Get rights and content

Abstract

1.
1. Chlorination of ovalbumin results in its enhanced immunogenic properties.
2.
2. This has been evaluated by the interleukine-2 production after incubation of the modified protein with antigen presenting cells and T helper cells.

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The ratio of CD8+/CD4+ T cells in the TME of combined group, associated with favorable prognosis after immunotherapy, also showed an increase compared to the anti-PD-1 alone, but not to the HOCl-CDS hydrogel alone (Fig. 6g-i). Although immunotherapies have changed melanoma treatment, not all patients respond [32–34]. More studies are trying to develop new strategies to potentiate cancer immunotherapy.
Autologous tumor cells and cell-derived secretions (CDS) can induce antitumor immune responses. The conditions in which cells are cultured and treated impact CDS, and cellular insults alter their composition and function. In this study, we generated CDS from tumor cells exposed to normal culture conditions, hypoxia, cisplatin, radiotherapy, photodynamic therapy, or hypochlorous acid (HOCl). In vitro HOCl-CDS showed the strongest stimulatory effects on dendritic cells and macrophages compared to CDS generated by hypoxia, cisplatin, radiotherapy or photodynamic therapy. To improve HOCl-CDS activity at the tumor site, we loaded HOCl-CDS into a melittin-encapsulated hydrogel scaffold. When injected intratumorally, the HOCl-CDS hydrogel promoted tumor cell death, cytotoxic T lymphocyte infiltration, and tumor-associated macrophage reprogramming towards an M1 phenotype. The hydrogel inhibited tumor growth and prolonged the survival of mice bearing B16–F10 melanoma. Furthermore, hydrogel-delivered HOCl-CDS augmented the antitumor effects of immune checkpoint blockade. These results underscore the importance of the CDS generation method and delivery approach for improving cancer immunotherapy.
The other myeloperoxidase: Emerging functions
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Several data indicate that the MPO-mediated antigen modifications could partially be responsible for these neutrophil actions as they can boost the immune response and suggest therefore that MPO can act as an adjuvant. Thus: (i) HOCl modification of ovalbumin improves its processing and presentation, increases the specific T cell response, and lowers the antigen amount required to stimulate the immune response [134,135]. ( ii) HOCl modification of lactalbumin induces a higher specific T cell activation [136]. (
Myeloperoxidase (MPO) is a member of the mammalian peroxidase family. It is mainly expressed in neutrophils, monocytes and macrophages. As a catalyzer of reactive oxidative species and radical species formation, it contributes to neutrophil bactericidal activity. Nevertheless MPO invalidation does not seem to have major health consequences in affected individuals. This suggests that MPO might have alternative functions supporting its conservation during evolution. We will review the available data supporting these non-canonical functions in terms of tissue specific expression, function and enzymatic activity. Thus, we discuss its cell type specific expression. We review in between others its roles in angiogenesis, endothelial (dys-) function, immune reaction, and inflammation. We summarize its pathological actions in clinical conditions such as cardiovascular disease and cancer.
Airway exposure to hypochlorite prior to ovalbumin induces airway hyperreactivity without evidence for allergic sensitization
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Firstly, chlorination products may induce epithelial hyperpermeability leading to an increased amount of allergen having access to the subepithelial dendritic cells and thereby amplifying sensitization in atopic subjects (Bernard, 2007). Allergens have also been shown to be very important targets for oxidants released when Cl2 or NaClO are added to water; e.g. ClO− is able to react with proteins, which could result in either breaking tolerance towards self-proteins or increasing immunogenicity creating new epitopes or revealing cryptic epitopes through protein unfolding (Olszowska et al., 1989; Marcinkiewicz et al., 1991; Westman and Harris, 2004). For other allergenic proteins however, such as Fel d 1 and Der p 1, the opposite could be observed; these proteins can be denatured by chlorine-based oxidants and in this way lose their immunogenicity (Chen and Eggleston, 2001; Matsui et al., 2003).
Some epidemiologic studies have indicated that attendance to chlorinated swimming pools is associated with airway hyperreactivity (AHR), allergies and asthma.
To investigate the effects of sodium hypochlorite (NaClO), the main pool disinfectant, on allergic sensitization and airway inflammation in mice.
In a first series of experiments, mice were sensitized to ovalbumin (OVA), followed by OVA aerosols with or without prior nasal instillation of NaClO (3 ppm active chlorine). In a second series, naïve mice received 1–7 nasal instillations of OVA, 10 min after instillations of NaClO or water. After 1, 3, 5 and 7 exposures airway reactivity to methacholine, cellular inflammation in bronchoalveolar lavage (BAL), serum OVA-specific IgEs and lung Th2 cytokines were measured.
In the first mouse model, airway allergy parameters were not significantly altered upon NaClO administration. However in the second model, NaClO exposure prior to OVA did induce AHR, already after 1 combined application. Combined NaClO + OVA exposure did not lead to an influx of inflammatory cells in BAL fluid or production of anti-OVA IgEs. No AHR developed when OVA was heat-denatured, pre-chlorinated, or replaced by bovine serum albumin or lipopolysaccharide.
Nasal instillation of NaClO prior to OVA induces AHR without allergic sensitization. This response is OVA-specific.
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Hypochlorous acid (HOCl) is a potent oxidant formed from hydrogen peroxide (H2O2) and chloride ion (Cl−) in a reaction catalyzed by the neutrophil enzyme myeloperoxidase (MPO) during acute inflammation. As well as showing strong microbicidal activity, HOCl has been shown to enhance the immunogenicity of protein antigens by several folds via oxidation in vivo and ex vivo[104–106]. It has been demonstrated that proteins oxidized by HOCl are more readily taken up and processed by APCs and led to enhanced activation of antigen-specific T cells in vitro[107–110].
Although cancer vaccines with defined antigens are commonly used, the use of whole tumor cell preparations in tumor immunotherapy is a very promising approach and can obviate some important limitations in vaccine development. Whole tumor cells are a good source of TAAs and can induce simultaneous CTLs and CD4⁺ T helper cell activation. We review current approaches to prepare whole tumor cell vaccines, including traditional methods of freeze–thaw lysates, tumor cells treated with ultraviolet irradiation, and RNA electroporation, along with more recent methods to increase tumor cell immunogenicity with HOCl oxidation or infection with replication-incompetent herpes simplex virus.
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Enhancement of immunogenic properties of ovalbumin as a result of its chlorination

Abstract

Biochem. biophys. Res. Commun.

Int. J. Biochem.

Biochim. biophys. Acta

Cell Immun.

Endopeptidase activities associated with the plasma membrane compartment of an antigen-presenting B cell

Clin. Exp. Immun.

The biochemistry and cell biology of antigen processing

Immun. Rev.