Monoclonal antibodies targeting surface exposed epitopes of Candida albicans cell wall proteins confer in vivo protection in an infection model

Soumya Palliyil; Mark Mawer; Sami  Alwafi; Lily  Fogg; Giuseppe Buda de Cesare; Tyng  Tan; Louise Walker; Donna  MacCalllum; Andrew Porter; Carol Munro

doi:10.1128/aac.01957-21

Monoclonal antibodies targeting surface exposed epitopes of Candida albicans cell wall proteins confer in vivo protection in an infection model

Soumya Palliyil^* (Corresponding Author), Mark Mawer, Sami Alwafi, Lily Fogg, Giuseppe Buda de Cesare, Tyng Tan, Louise Walker, Donna MacCalllum, Andrew Porter, Carol Munro

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

Monoclonal antibody (mAb)-based immunotherapies targeting systemic and deep-seated fungal infections are still in their early stages of development, with no licensed antifungal mAbs currently being available for patients at risk. The cell wall glycoproteins of Candida albicans are of particular interest as potential targets for therapeutic antibody generation due to their extracellular location and key involvement in fungal pathogenesis. Here, we describe the generation of recombinant human antibodies specifically targeting two key cell wall proteins (CWPs) in C. albicans: Utr2 and Pga31. These antibodies were isolated from a phage display antibody library using peptide antigens representing the surface-exposed regions of CWPs expressed at elevated levels during in vivo infection. Reformatted human-mouse chimeric mAbs preferentially recognized C. albicans hyphal forms compared to yeast cells, and increased binding was observed when the cells were grown in the presence of the antifungal agent caspofungin. In J774.1 macrophage interaction assays, mAb pretreatment resulted in the faster engulfment of C. albicans cells, suggesting a role of the CWP antibodies as opsonizing agents during phagocyte recruitment. Finally, in a series of clinically predictive mouse models of systemic candidiasis, our lead mAb achieved improved survival (83%) and a several-log reduction of the fungal burden in the kidneys, similar to the levels achieved for the fungicidal drug caspofungin and superior to the therapeutic efficacy of any anti-Candida mAb reported to date.

Original language	English
Article number	e01957-21
Journal	Antimicrobial Agents and Chemotherapy
Volume	66
Issue number	4
Early online date	14 Mar 2022
DOIs	https://doi.org/10.1128/aac.01957-21
Publication status	Published - 19 Apr 2022

Bibliographical note

ACKNOWLEDGMENTS
We gratefully acknowledge Kevin McKenzie and Lucy Wight from the University of Aberdeen Microscopy and Histology Facility for training and access to fluorescence microscopy and for their support and assistance in this work. We also gratefully acknowledge David Stead from Aberdeen Proteomics for his support and assistance with the Candida proteome analysis and the staff of the University of Aberdeen Medical Research Facility for their support and assistance with the mouse studies.
This work was supported by the following research grants: the High Throughput and Fragment Screening Fund, Scottish Universities Life Sciences Alliance (SULSA); a seed corn award from the University of Aberdeen Wellcome Trust Institutional Strategic Support Fund; an M.Res. studentship by the Medical Research Council Centre for Medical Mycology at the University of Aberdeen (grant number MR/P501955/1); a Ph.D. studentship from the Institute of Medical Sciences, University of Aberdeen; a Ph.D. studentship from Taibah University and a Saudi Government scholarship; and a Ph.D. studentship by the European Union’s Horizon 2020 research and innovation program under Marie Sklodowska-Curie grant agreement number H2020-MSCA-ITN-2014-642095 (OPATHY).
C.A.M., S.P., and A.J.P. contributed to the concept and study design. C.A.M. and S.P. developed the methodology. S.A.A. and L.F. performed recombinant antibody generation and ELISAs. L.F. and M.M. completed IgG reformatting and the production of mAbs for animal studies. M.M., T.H.T., and L.A.W. performed ELISAs and immunofluorescence staining. M.M. performed macrophage assays, and D.M.M. planned, conducted, and analyzed animal studies. C.A.M., S.P., and A.J.P. contributed to funding acquisition and project administration, and C.A.M., S.P., and L.A.W. contributed to the supervision and training of M.Res. and Ph.D. students. S.P. wrote the original draft, and C.A.M., D.M.M., and A.J.P. completed review and editing. All authors had full access to the data and approved the manuscript before it was submitted by the corresponding author(s).
S.P., A.J.P., and C.A.M. are inventors on a patent related to the development of antifungal antibodies to surface-exposed epitopes of fungal pathogens owned by the University of Aberdeen. All other authors declare that they have no competing interests.

Data Availability Statement

All data are available in the text or the supplemental material. Antibodies described in this paper will be available for research purposes through a material transfer agreement with the University of Aberdeen.

Keywords

Candida albicans
anti-Candida mAbs
immunotherapy
monoclonal antibodies

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Palliyil, S., Mawer, M., Alwafi, S., Fogg, L., Buda de Cesare, G., Tan, T., Walker, L., MacCalllum, D., Porter, A., & Munro, C. (2022). Monoclonal antibodies targeting surface exposed epitopes of Candida albicans cell wall proteins confer in vivo protection in an infection model. Antimicrobial Agents and Chemotherapy, 66(4), Article e01957-21. https://doi.org/10.1128/aac.01957-21

Monoclonal antibodies targeting surface exposed epitopes of Candida albicans cell wall proteins confer in vivo protection in an infection model. / Palliyil, Soumya (Corresponding Author); Mawer, Mark; Alwafi, Sami et al.
In: Antimicrobial Agents and Chemotherapy, Vol. 66, No. 4, e01957-21, 19.04.2022.

Research output: Contribution to journal › Article › peer-review

Palliyil, S, Mawer, M, Alwafi, S, Fogg, L, Buda de Cesare, G, Tan, T, Walker, L, MacCalllum, D, Porter, A & Munro, C 2022, 'Monoclonal antibodies targeting surface exposed epitopes of Candida albicans cell wall proteins confer in vivo protection in an infection model', Antimicrobial Agents and Chemotherapy, vol. 66, no. 4, e01957-21. https://doi.org/10.1128/aac.01957-21

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title = "Monoclonal antibodies targeting surface exposed epitopes of Candida albicans cell wall proteins confer in vivo protection in an infection model",

abstract = "Monoclonal antibody (mAb)-based immunotherapies targeting systemic and deep-seated fungal infections are still in their early stages of development, with no licensed antifungal mAbs currently being available for patients at risk. The cell wall glycoproteins of Candida albicans are of particular interest as potential targets for therapeutic antibody generation due to their extracellular location and key involvement in fungal pathogenesis. Here, we describe the generation of recombinant human antibodies specifically targeting two key cell wall proteins (CWPs) in C. albicans: Utr2 and Pga31. These antibodies were isolated from a phage display antibody library using peptide antigens representing the surface-exposed regions of CWPs expressed at elevated levels during in vivo infection. Reformatted human-mouse chimeric mAbs preferentially recognized C. albicans hyphal forms compared to yeast cells, and increased binding was observed when the cells were grown in the presence of the antifungal agent caspofungin. In J774.1 macrophage interaction assays, mAb pretreatment resulted in the faster engulfment of C. albicans cells, suggesting a role of the CWP antibodies as opsonizing agents during phagocyte recruitment. Finally, in a series of clinically predictive mouse models of systemic candidiasis, our lead mAb achieved improved survival (83%) and a several-log reduction of the fungal burden in the kidneys, similar to the levels achieved for the fungicidal drug caspofungin and superior to the therapeutic efficacy of any anti-Candida mAb reported to date.",

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author = "Soumya Palliyil and Mark Mawer and Sami Alwafi and Lily Fogg and {Buda de Cesare}, Giuseppe and Tyng Tan and Louise Walker and Donna MacCalllum and Andrew Porter and Carol Munro",

note = "ACKNOWLEDGMENTS We gratefully acknowledge Kevin McKenzie and Lucy Wight from the University of Aberdeen Microscopy and Histology Facility for training and access to fluorescence microscopy and for their support and assistance in this work. We also gratefully acknowledge David Stead from Aberdeen Proteomics for his support and assistance with the Candida proteome analysis and the staff of the University of Aberdeen Medical Research Facility for their support and assistance with the mouse studies. This work was supported by the following research grants: the High Throughput and Fragment Screening Fund, Scottish Universities Life Sciences Alliance (SULSA); a seed corn award from the University of Aberdeen Wellcome Trust Institutional Strategic Support Fund; an M.Res. studentship by the Medical Research Council Centre for Medical Mycology at the University of Aberdeen (grant number MR/P501955/1); a Ph.D. studentship from the Institute of Medical Sciences, University of Aberdeen; a Ph.D. studentship from Taibah University and a Saudi Government scholarship; and a Ph.D. studentship by the European Union{\textquoteright}s Horizon 2020 research and innovation program under Marie Sklodowska-Curie grant agreement number H2020-MSCA-ITN-2014-642095 (OPATHY). C.A.M., S.P., and A.J.P. contributed to the concept and study design. C.A.M. and S.P. developed the methodology. S.A.A. and L.F. performed recombinant antibody generation and ELISAs. L.F. and M.M. completed IgG reformatting and the production of mAbs for animal studies. M.M., T.H.T., and L.A.W. performed ELISAs and immunofluorescence staining. M.M. performed macrophage assays, and D.M.M. planned, conducted, and analyzed animal studies. C.A.M., S.P., and A.J.P. contributed to funding acquisition and project administration, and C.A.M., S.P., and L.A.W. contributed to the supervision and training of M.Res. and Ph.D. students. S.P. wrote the original draft, and C.A.M., D.M.M., and A.J.P. completed review and editing. All authors had full access to the data and approved the manuscript before it was submitted by the corresponding author(s). S.P., A.J.P., and C.A.M. are inventors on a patent related to the development of antifungal antibodies to surface-exposed epitopes of fungal pathogens owned by the University of Aberdeen. All other authors declare that they have no competing interests.",

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AU - Mawer, Mark

AU - Alwafi, Sami

AU - Fogg, Lily

AU - Buda de Cesare, Giuseppe

AU - Tan, Tyng

AU - Walker, Louise

AU - MacCalllum, Donna

AU - Porter, Andrew

AU - Munro, Carol

N1 - ACKNOWLEDGMENTS We gratefully acknowledge Kevin McKenzie and Lucy Wight from the University of Aberdeen Microscopy and Histology Facility for training and access to fluorescence microscopy and for their support and assistance in this work. We also gratefully acknowledge David Stead from Aberdeen Proteomics for his support and assistance with the Candida proteome analysis and the staff of the University of Aberdeen Medical Research Facility for their support and assistance with the mouse studies. This work was supported by the following research grants: the High Throughput and Fragment Screening Fund, Scottish Universities Life Sciences Alliance (SULSA); a seed corn award from the University of Aberdeen Wellcome Trust Institutional Strategic Support Fund; an M.Res. studentship by the Medical Research Council Centre for Medical Mycology at the University of Aberdeen (grant number MR/P501955/1); a Ph.D. studentship from the Institute of Medical Sciences, University of Aberdeen; a Ph.D. studentship from Taibah University and a Saudi Government scholarship; and a Ph.D. studentship by the European Union’s Horizon 2020 research and innovation program under Marie Sklodowska-Curie grant agreement number H2020-MSCA-ITN-2014-642095 (OPATHY). C.A.M., S.P., and A.J.P. contributed to the concept and study design. C.A.M. and S.P. developed the methodology. S.A.A. and L.F. performed recombinant antibody generation and ELISAs. L.F. and M.M. completed IgG reformatting and the production of mAbs for animal studies. M.M., T.H.T., and L.A.W. performed ELISAs and immunofluorescence staining. M.M. performed macrophage assays, and D.M.M. planned, conducted, and analyzed animal studies. C.A.M., S.P., and A.J.P. contributed to funding acquisition and project administration, and C.A.M., S.P., and L.A.W. contributed to the supervision and training of M.Res. and Ph.D. students. S.P. wrote the original draft, and C.A.M., D.M.M., and A.J.P. completed review and editing. All authors had full access to the data and approved the manuscript before it was submitted by the corresponding author(s). S.P., A.J.P., and C.A.M. are inventors on a patent related to the development of antifungal antibodies to surface-exposed epitopes of fungal pathogens owned by the University of Aberdeen. All other authors declare that they have no competing interests.

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N2 - Monoclonal antibody (mAb)-based immunotherapies targeting systemic and deep-seated fungal infections are still in their early stages of development, with no licensed antifungal mAbs currently being available for patients at risk. The cell wall glycoproteins of Candida albicans are of particular interest as potential targets for therapeutic antibody generation due to their extracellular location and key involvement in fungal pathogenesis. Here, we describe the generation of recombinant human antibodies specifically targeting two key cell wall proteins (CWPs) in C. albicans: Utr2 and Pga31. These antibodies were isolated from a phage display antibody library using peptide antigens representing the surface-exposed regions of CWPs expressed at elevated levels during in vivo infection. Reformatted human-mouse chimeric mAbs preferentially recognized C. albicans hyphal forms compared to yeast cells, and increased binding was observed when the cells were grown in the presence of the antifungal agent caspofungin. In J774.1 macrophage interaction assays, mAb pretreatment resulted in the faster engulfment of C. albicans cells, suggesting a role of the CWP antibodies as opsonizing agents during phagocyte recruitment. Finally, in a series of clinically predictive mouse models of systemic candidiasis, our lead mAb achieved improved survival (83%) and a several-log reduction of the fungal burden in the kidneys, similar to the levels achieved for the fungicidal drug caspofungin and superior to the therapeutic efficacy of any anti-Candida mAb reported to date.

AB - Monoclonal antibody (mAb)-based immunotherapies targeting systemic and deep-seated fungal infections are still in their early stages of development, with no licensed antifungal mAbs currently being available for patients at risk. The cell wall glycoproteins of Candida albicans are of particular interest as potential targets for therapeutic antibody generation due to their extracellular location and key involvement in fungal pathogenesis. Here, we describe the generation of recombinant human antibodies specifically targeting two key cell wall proteins (CWPs) in C. albicans: Utr2 and Pga31. These antibodies were isolated from a phage display antibody library using peptide antigens representing the surface-exposed regions of CWPs expressed at elevated levels during in vivo infection. Reformatted human-mouse chimeric mAbs preferentially recognized C. albicans hyphal forms compared to yeast cells, and increased binding was observed when the cells were grown in the presence of the antifungal agent caspofungin. In J774.1 macrophage interaction assays, mAb pretreatment resulted in the faster engulfment of C. albicans cells, suggesting a role of the CWP antibodies as opsonizing agents during phagocyte recruitment. Finally, in a series of clinically predictive mouse models of systemic candidiasis, our lead mAb achieved improved survival (83%) and a several-log reduction of the fungal burden in the kidneys, similar to the levels achieved for the fungicidal drug caspofungin and superior to the therapeutic efficacy of any anti-Candida mAb reported to date.

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Monoclonal antibodies targeting surface exposed epitopes of Candida albicans cell wall proteins confer in vivo protection in an infection model

Abstract

Bibliographical note

Data Availability Statement

Keywords

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