Extraction, characterization and biological activity of a (1,3)(1,6)-β-d-glucan from the pathogenic oomycete Pythium insidiosum

Tondolo, Juliana S. M.; Ledur, Pauline Christ; Loreto, Érico S.; Verdi, Camila Marina; Bitencourt, Paula Eliete Rodrigues; Jesus, Francielli Pantella Kunz de; Rocha, Jean P.; Alves, Sydney Hartz; Sassaki, Guilherme L.; Santúrio, Jânio Morais

doi:10.1016/j.carbpol.2016.10.053

Cited by 22 publications

(13 citation statements)

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“…Lymphocytes from all groups were cultivated with RPMI-1640 (non-stimulated cells: N/S) or were stimulated with a P . insidiosum cell wall β-glucan (Glucan, as a mitogen) at 0.1 mg mL -1 [23], and concanavalin A (ConA, Sigma-Aldrich) was used as the mitogen positive control at 0.04 mg mL -1 .…”

Section: Methodsmentioning

confidence: 99%

Chemically induced disseminated pythiosis in BALB/c mice: A new experimental model for Pythium insidiosum infection

et al. 2017

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Pythiosis is a severe and life-threatening disease that affects humans and various animal species. We report a model of vascular/disseminated pythiosis occurring after subcutaneous inoculation of 2 x 104 Pythium insidiosum zoospores/mL in immunocompromised BALB/c mice. For this model, we carried out two rounds of experiments. First, we evaluated two protocols of immunosuppression before inoculation: cyclophosphamide at 150 mg/kg (CYP group) and cyclophosphamide 200 mg/kg plus hydrocortisone acetate at 250 mg/kg (CYP+HCA group). It was not possible to obtain mortality in the CYP group; however, the combination of CYP+HCA altered disease outcomes, with mortality rates reaching 60%. Second, we used the CYP+HCA immunosuppression protocol to analyze the histological and immunological statuses triggered by disease. When we inoculated immunocompetent mice with P. insidiosum zoospores, self-healing occurred via increased levels of IL-2, IFN-γ and IL-17A, which are characteristic of the Th1/Th17 cytokine response. For infected and immunosuppressed mice, the cytokine profiles showed high levels of IL-10, IL-6 and TNF-α. Increased IL-10 values are related to fungal infection susceptibility and led us to speculate that infection may be established through suppression of the host immune response. In addition, histopathological evaluation of the kidneys and liver demonstrated the presence of hyphae and the cellular findings suggested an acute vascular inflammation that mimics vascular/disseminated pythiosis in humans. This is the first murine model for pythiosis that is useful both for understanding the pathogenesis of this disease and for evaluating new treatment approaches.

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Section: Methodsmentioning

confidence: 99%

Chemically induced disseminated pythiosis in BALB/c mice: A new experimental model for Pythium insidiosum infection

et al. 2017

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“…However, recovery of the PIA-immunized humans and animals from pythiosis is associated with switching Th2 to T helper-1 (Th1) mediated immunity [131,142,244,251,252]. An APC should process an immunomodulating antigen of P. insidiosum, such as (1,3)(1,6)-β-glucan, which is a main cell wall component [251,253,254], before priming a resulting antigenic epitope to a naïve T cell (via MHC-TCR complex) [242,243]. This cellular interaction might trigger the release of IFNγ and IL-2 for enhancing the differentiation of Th0 to Th1 cell and Th0 to regulatory T (Treg) cell, respectively [242,246,251,252,255,256].…”

Section: Proposed Mechanism Of P Insidiosum Antigen-based Immunotherapymentioning

confidence: 99%

“…This cellular interaction might trigger the release of IFNγ and IL-2 for enhancing the differentiation of Th0 to Th1 cell and Th0 to regulatory T (Treg) cell, respectively [242,246,251,252,255,256]. The Th1 cell produces IFN-γ to activate the macrophages and cytotoxic T lymphocytes [131,142,242,244,246,251,253,256]. The PIA raises the T helper-17 (Th17) cells, which secrete IL-17A to recruit neutrophils and macrophages into the infected tissue [242,243,246,[251][252][253].…”

Section: Proposed Mechanism Of P Insidiosum Antigen-based Immunotherapymentioning

confidence: 99%

“…The Th1 cell produces IFN-γ to activate the macrophages and cytotoxic T lymphocytes [131,142,242,244,246,251,253,256]. The PIA raises the T helper-17 (Th17) cells, which secrete IL-17A to recruit neutrophils and macrophages into the infected tissue [242,243,246,[251][252][253]. The PIA could also promote the release of the IL-10 cytokine, which relates to the anti-inflammatory and immunoregulation activity of Treg cells [242,246,[251][252][253]256].…”

Section: Proposed Mechanism Of P Insidiosum Antigen-based Immunotherapymentioning

confidence: 99%

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History and Perspective of Immunotherapy for Pythiosis

Yolanda

Krajaejun

2021

Vaccines

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The fungus-like microorganism Pythium insidiosum causes pythiosis, a life-threatening infectious disease increasingly reported worldwide. Antimicrobial drugs are ineffective. Radical surgery is an essential treatment. Pythiosis can resume post-surgically. Immunotherapy using P. insidiosum antigens (PIA) has emerged as an alternative treatment. This review aims at providing up-to-date information of the immunotherapeutic PIA, with the focus on its history, preparation, clinical application, outcome, mechanism, and recent advances, in order to promote the proper use and future development of this treatment modality. P. insidiosum crude extract is the primary source of immunotherapeutic antigens. Based on 967 documented human and animal (mainly horses) pythiosis cases, PIA immunotherapy reduced disease morbidity and mortality. Concerning clinical outcomes, 19.4% of PIA-immunized human patients succumbed to vascular pythiosis instead of 41.0% in unimmunized cases. PIA immunotherapy may not provide an advantage in a local P. insidiosum infection of the eye. Both PIA-immunized and unimmunized horses with pythiosis showed a similar survival rate of ~70%; however, demands for surgical intervention were much lesser in the immunized cases (22.8% vs. 75.2%). The proposed PIA action involves switching the non-protective T-helper-2 to protective T-helper-1 mediated immunity. By exploring the available P. insidiosum genome data, synthetic peptides, recombinant proteins, and nucleic acids are potential sources of the immunotherapeutic antigens worth investigating. The PIA therapeutic property needs improvement for a better prognosis of pythiosis patients.

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“…Unlike fungi, the cell wall of oomycetes primarily composes of cellulose, glucans, and to a lesser extent, chitin (M elida et al, 2013). β-glucan is a primary carbohydrate found in the P. insidiosum extract (Tondolo et al, 2017). The β-glucanase Exo1 may involve in cell wall remodeling, hyphal growth, and invasion of P. insidiosum (Keeratijarut et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Expression, purification, and characterization of the recombinant exo-1,3-β-glucanase (Exo1) of the pathogenic oomycete Pythium insidiosum

Rotchanapreeda

Kumsang

Sae‐Chew

et al. 2020

Heliyon

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Pythiosis is a deadly infectious disease of humans and animals living in tropical and subtropical countries. The causative agent is the oomycete Pythium insidiosum. Treatment of pythiosis is challenging. The use of antimicrobial agents usually fails in the treatment of pythiosis. Many patients undergo surgical removal of an infected organ (i.e., eye, arm, and leg). The immunotherapeutic vaccine, prepared from the crude extract of P. insidiosum, shows limited efficacy against pythiosis. The fatal outcome occurs in patients with advanced disease. There are urgent needs for an effective therapeutic modality for pythiosis. Recently, the exo-1,3-β-glucanase (Exo1) has been identified as a conserve immunoreactive protein of P. insidiosum. Exo1 was predicted to reside at the cell membrane and hydrolyze cell wall β-glucan during cell growth. An Exo1 ortholog is absent in the human genome, making it an appealing target for drug or vaccine development. We attempted to clone and express the codonoptimized exo1 gene of P. insidiosum in E. coli. To solve the inclusion body formation, expression and purification of Exo1 were achievable in the denaturing condition using SDS-and urea-based buffers. Exo1 lacked hydrolytic activity due to the absence of proper protein folding and post-translational modifications. ELISA and Western blot analyses demonstrated the immunoreactivity of Exo1 against pythiosis sera. In conclusion, we successfully expressed and purified the immunoreactive Exo1 protein of P. insidiosum. The recombinant Exo1 can be produced at an unlimited amount and could serve as an extra protein to enhance the effectiveness of the current form of the vaccine against pythiosis.

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Extraction, characterization and biological activity of a (1,3)(1,6)-β-d-glucan from the pathogenic oomycete Pythium insidiosum

Cited by 22 publications

References 33 publications

Chemically induced disseminated pythiosis in BALB/c mice: A new experimental model for Pythium insidiosum infection

Chemically induced disseminated pythiosis in BALB/c mice: A new experimental model for Pythium insidiosum infection

History and Perspective of Immunotherapy for Pythiosis

Expression, purification, and characterization of the recombinant exo-1,3-β-glucanase (Exo1) of the pathogenic oomycete Pythium insidiosum

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