Subtilisin 6 From the Dermatophyte Trichophyton benhamiae Is a Marker of Infection but Not a Unique Virulence Factor.

Trichophyton benhamiae; dermatophytosis; fungal marker; infection model; subtilisin 6; Virulence Factors; Subtilisins; Fungal Proteins; Animals; Mice; Virulence; Skin/microbiology; Skin/pathology; Subtilisins/genetics; Female; Fungal Proteins/genetics; Fungal Proteins/metabolism; Gene Deletion; Mice, Inbred BALB C; Virulence Factors/genetics; Disease Models, Animal; Trichophyton/genetics; Trichophyton/pathogenicity; Tinea/microbiology

Abstract :

[en] [en] BACKGROUND: Trichophyton benhamiae is a common dermatophyte whose natural host is the guinea pig and which causes highly inflammatory skin lesions in humans. The subtilisin 6 (SUB6) of this fungus belongs to a family of 12 SUB genes. Its encoding gene, overexpressed in vivo but not in vitro, has been considered a potentially important virulence factor, but its role in pathogenesis remains to be elucidated. OBJECTIVES: The aim of this study was to assess the role of T. benhamiae SUB6 in virulence in a mouse skin infection model. METHODS: To assess the contribution of SUB6 to virulence, SUB6-deleted (ΔSUB6) and complemented strains were generated by genetic transformation. The pathogenicity of these strains was compared with that of the parental strain in vivo in mice, based on the evolution of skin symptoms, histopathological lesions and molecular analyses targeting the expression of host pro-inflammatory genes and fungal genes encoding subtilisins from the same family as SUB6. RESULTS: The ΔSUB6 strain induced superficial skin signs and histopathological inflammatory lesions similar to those caused by the parental strain. Significant overexpression of the SUB1, SUB3, SUB8 and SUB10 genes in the tissues was observed regardless of the strain tested, with no difference between these strains, reflecting the absence of any compensatory mechanism among subtilisins. CONCLUSIONS: SUB6 appears to be more of a marker of fungal infection than a virulence factor, at least acting alone.

Disciplines :

Veterinary medicine & animal health

Author, co-author :

Poirier, Wilfried ; Université de Liège - ULiège > Fundamental and Applied Research for Animals and Health (FARAH) > FARAH: Santé publique vétérinaire

Faway, Émilie ; Molecular Physiology Research Unit, NAmur Research Institute for Life Sciences (URPHYM-NARILIS), Faculty of Medicine, University of Namur, Namur, Belgium

Yamada, Tsuyoshi ; Institute of Medical Mycology, Teikyo University, Tokyo, Japan ; Asia International Institute of Infectious Disease Control, Teikyo University, Tokyo, Japan

Ozawa, Kiyotaka ; Institute of Medical Mycology, Teikyo University, Tokyo, Japan ; Medical Mycology Research Center, Chiba University, Chiba, Japan

Maréchal, Françoise ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires (DMI)

Monod, Michel ; Department of Dermatology, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland

Poumay, Yves ; Molecular Physiology Research Unit, NAmur Research Institute for Life Sciences (URPHYM-NARILIS), Faculty of Medicine, University of Namur, Namur, Belgium

Mignon, Bernard ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires (DMI) > Parasitologie et pathologie des maladies parasitaires

Language :

English

Title :

Subtilisin 6 From the Dermatophyte Trichophyton benhamiae Is a Marker of Infection but Not a Unique Virulence Factor.

Publication date :

February 2025

Journal title :

Mycoses

ISSN :

0933-7407

eISSN :

1439-0507

Publisher :

Wiley, Germany

Volume :

Issue :

Pages :

e70037

Peer reviewed :

Peer Reviewed verified by ORBi

Additional URL :

https://onlinelibrary.wiley.com/doi/pdf/10.1111/myc.70037

Available on ORBi :

since 28 January 2026

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Bibliography

B. Havlickova, V. A. Czaika, and M. Friedrich, “Epidemiological Trends in Skin Mycoses Worldwide,” Mycoses 51, no. 4 (2008): 2–15, https://doi.org/10.1111/j.1439-0507.2008.01606.x.
A. D. Paryuni, S. Indarjulianto, and S. Widyarini, “Dermatophytosis in Companion Animals: A Review,” Vet World 13, no. 6 (2020): 1174–1181, https://doi.org/10.14202/vetworld.2020.1174-1181.
P. Zhan and W. Liu, “The Changing Face of Dermatophytic Infections Worldwide,” Mycopathologia 182, no. 1–2 (2017): 77–86, https://doi.org/10.1007/s11046-016-0082-8.
D. Sriranganadane, P. Waridel, K. Salamin, et al., “Identification of Novel Secreted Proteases During Extracellular Proteolysis by Dermatophytes at Acidic pH,” Proteomics 11, no. 22 (2011): 4422–4433, https://doi.org/10.1002/pmic.201100234.
N. M. Martinez-Rossi, N. T. A. Peres, and A. Rossi, “Pathogenesis of Dermatophytosis: Sensing the Host Tissue,” Mycopathologia 182, no. 1–2 (2017): 215–227, https://doi.org/10.1007/s11046-016-0057-9.
S. Vermout, J. Tabart, A. Baldo, A. Mathy, B. Losson, and B. Mignon, “Pathogenesis of Dermatophytosis,” Mycopathologia 166, no. 5–6 (2008): 267–275, https://doi.org/10.1007/s11046-008-9104-5.
E. T. Băguţ, A. Baldo, A. Mathy, et al., “Subtilisin Sub3 Is Involved in Adherence of Microsporum canis to Human and Animal Epidermis,” Veterinary Microbiology 160, no. 3-4 (2012): 413–419, https://doi.org/10.1016/j.vetmic.2012.06.011.
A. Baldo, M. Monod, A. Mathy, et al., “Mechanisms of Skin Adherence and Invasion by Dermatophytes,” Mycoses 55, no. 3 (2012): 218–223, https://doi.org/10.1111/j.1439-0507.2011.02081.x.
A. Baldo, A. Mathy, J. Tabart, et al., “Secreted Subtilisin Sub3 From Microsporum canis Is Required for Adherence to but Not for Invasion of the Epidermis,” British Journal of Dermatology 162, no. 5 (2010): 990–997, https://doi.org/10.1111/j.1365-2133.2009.09608.x.
A. Baldo, J. Tabart, S. Vermout, et al., “Secreted Subtilisins of Microsporum canis Are Involved in Adherence of Arthroconidia to Feline Corneocytes,” Journal of Medical Microbiology 57, no. 9 (2008): 1152–1156, https://doi.org/10.1099/jmm.0.47827-0.
M. Fratti, O. Bontems, K. Salamin, E. Guenova, and M. Monod, “Survey on Dermatophytes Isolated From Animals in Switzerland in the Context of the Prevention of Zoonotic Dermatophytosis,” Journal of Fungi 9, no. 2 (2023): 253, https://doi.org/10.3390/jof9020253.
J. Fumeaux, M. Mock, B. Ninet, et al., “First Report of Arthroderma benhamiae in Switzerland,” Dermatology 208, no. 3 (2004): 244–250, https://doi.org/10.1159/000077311.
V. D. T. Tran, N. De Coi, M. Feuermann, et al., “RNA Sequencing-Based Genome Reannotation of the Dermatophyte Arthroderma benhamiae and Characterization of Its Secretome and Whole Gene Expression Profile During Infection,” mSystems 1, no. 4 (2016): e0003616, https://doi.org/10.1128/mSystems.00036-16.
E. Faway, W. Poirier, F. Maréchal, Y. Poumay, and B. Mignon, “Expression of Fungal and Host Markers in Models of Dermatophytosis on Mice and Human Epidermis,” Journal of Investigative Dermatology (2024), https://doi.org/10.1016/j.jid.2024.08.010.
J. A. Woodfolk, L. M. Wheatley, R. V. Piyasena, D. C. Benjamin, and T. A. E. Platts-Mills, “Trichophyton Antigens Associated With IgE Antibodies and Delayed Type Hypersensitivity: Sequence Homology to Two Families of Serine Proteinases,” Journal of Biological Chemistry 273, no. 45 (1998): 29489–29496, https://doi.org/10.1074/jbc.273.45.29489.
B. Méhul, P. Grundt, A. Genette, J. J. Voegel, and M. Monod, “Detection of Trichophyton rubrum and Trichophyton interdigitale in Onychomycosis Using Monoclonal Antibodies Against Sub6 (Tri r 2),” Mycoses 62, no. 1 (2019): 32–40, https://doi.org/10.1111/myc.12843.
B. Méhul, Z. Gu, A. Jomard, G. Laffet, M. Feuilhade, and M. Monod, “Sub6 (Tri r 2), an Onychomycosis Marker Revealed by Proteomics Analysis of Trichophyton rubrum Secreted Proteins in Patient Nail Samples,” Journal of Investigative Dermatology 136, no. 1 (2016): 331–333, https://doi.org/10.1038/JID.2015.367.
C. M. Baumbach, J. K. Michler, P. Nenoff, S. Uhrlaß, and W. Schrödl, “Visualising Virulence Factors: Trichophyton benhamiae Subtilisins Demonstrated in a Guinea Pig Skin Ex Vivo Model,” Mycoses 63, no. 9 (2020): 970–978, https://doi.org/10.1111/myc.13136.
J. Lindenhahn, T. Bartosch, C. M. Baumbach, et al., “Detection of Subtilisin 3 and 6 in Skin Biopsies of Cattle With Clinically Manifested Bovine Ringworm,” Medical Mycology 59, no. 3 (2021): 305–308, https://doi.org/10.1093/mmy/myaa089.
A. Burmester, E. Shelest, G. Glöckner, et al., “Comparative and Functional Genomics Provide Insights Into the Pathogenicity of Dermatophytic Fungi,” Genome Biology 12, no. 1 (2011): R7, https://doi.org/10.1186/gb-2011-12-1-r7.
M. Grumbt, V. Defaweux, B. Mignon, et al., “Targeted Gene Deletion and In Vivo Analysis of Putative Virulence Gene Function in the Pathogenic Dermatophyte Arthroderma benhamiae,” Eukaryotic Cell 10, no. 6 (2011): 842–853, https://doi.org/10.1128/EC.00273-10.
M. Grumbt, M. Monod, T. Yamada, C. Hertweck, J. Kunert, and P. Staib, “Keratin Degradation by Dermatophytes Relies on Cysteine Dioxygenase and a Sulfite Efflux Pump,” Journal of Investigative Dermatology 133, no. 6 (2013): 1550–1555, https://doi.org/10.1038/jid.2013.41.
T. Yamada, K. Makimura, K. Satoh, Y. Umeda, Y. Ishihara, and S. Abe, “Agrobacterium tumefaciens-Mediated Transformation of the Dermatophyte, Trichophyton mentagrophytes: An Efficient Tool for Gene Transfer,” Medical Mycology 47, no. 5 (2009): 485–494, https://doi.org/10.1080/13693780802322240.
H. Girardin and J. P. Latgé, “DNA Extraction and Quantitation,” in Molecular Biology of Pathogenic Fungi: A Laboratory Manual, eds. B. Maresca and G. S. Kobayashi (Telos Press, 1994), 5–9.
T. Yamada, M. Maeda, M. M. Alshahni, et al., “Terbinafine Resistance of Trichophyton Clinical Isolates Caused by Specific Point Mutations in the Squalene Epoxidase Gene,” Antimicrobial Agents and Chemotherapy 61, no. 7 (2017): e00115-17, https://doi.org/10.1128/AAC.00115-17.
E. Faway, C. Staerck, C. Danzelle, et al., “Towards a Standardized Procedure for the Production of Infective Spores to Study the Pathogenesis of Dermatophytosis,” Journal of Fungi 7, no. 12 (2021): 1029, https://doi.org/10.3390/jof7121029.
K. J. Livak and T. D. Schmittgen, “Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2(-Delta Delta C(T)) Method,” Methods San Diego Calif 25, no. 4 (2001): 402–408, https://doi.org/10.1006/meth.2001.1262.
Y. Shi, Q. Niu, X. Yu, et al., “Assessment of the Function of SUB6 in the Pathogenic Dermatophyte Trichophyton mentagrophytes,” Medical Mycology 54, no. 1 (2016): 59–71, https://doi.org/10.1093/mmy/myv071.
S. O. Suh, K. M. Grosso, and M. E. Carrion, “Multilocus Phylogeny of the Trichophyton mentagrophytes Species Complex and the Application of Matrix-Assisted Laser Desorption/Ionization-Time-Of-Flight (MALDI-TOF) Mass Spectrometry for the Rapid Identification of Dermatophytes,” Mycologia 110, no. 1 (2018): 118–130, https://doi.org/10.1080/00275514.2018.1443653.
B. Hube, R. Hay, J. Brasch, S. Veraldi, and M. Schaller, “Dermatomycoses and Inflammation: The Adaptive Balance Between Growth, Damage, and Survival,” Journal de Mycologie Médicale 25, no. 1 (2015): 44–58, https://doi.org/10.1016/j.mycmed.2014.11.002.
B. Mignon, J. Tabart, A. Baldo, A. Mathy, B. Losson, and S. Vermout, “Immunization and Dermatophytes,” Current Opinion in Infectious Diseases 21, no. 2 (2008): 134–140, https://doi.org/10.1097/QCO.0b013e3282f55de6.
V. L. Burstein, L. Guasconi, I. Beccacece, et al., “IL-17-Mediated Immunity Controls Skin Infection and T Helper 1 Response During Experimental Microsporum canis Dermatophytosis,” Journal of Investigative Dermatology 138, no. 8 (2018): 1744–1753, https://doi.org/10.1016/j.jid.2018.02.042.
M. P. Heinen, L. Cambier, N. Antoine, et al., “Th1 and Th17 Immune Responses Act Complementarily to Optimally Control Superficial Dermatophytosis,” Journal of Investigative Dermatology 139, no. 3 (2019): 626–637, https://doi.org/10.1016/j.jid.2018.07.040.
M. P. Heinen, L. Cambier, L. Fievez, and B. Mignon, “Are Th17 Cells Playing a Role in Immunity to Dermatophytosis?,” Mycopathologia 182, no. 1–2 (2017): 251–261, https://doi.org/10.1007/s11046-016-0093-5.
B. Hube and J. Naglik, “Candida albicans Proteinases: Resolving the Mystery of a Gene Family,” Microbiology (Reading, England) 147, no. Pt 8 (2001): 1997–2005, https://doi.org/10.1099/00221287-147-8-1997.
K. Jaton-Ogay, S. Paris, M. Huerre, et al., “Cloning and Disruption of the Gene Encoding an Extracellular Metalloprotease of Aspergillus fumigatus,” Molecular Microbiology 14, no. 5 (1994): 917–928, https://doi.org/10.1111/j.1365-2958.1994.tb01327.x.
X. Zhang, Y. Wang, W. Chi, et al., “Metalloprotease Genes of Trichophyton mentagrophytes Are Important for Pathogenicity,” Medical Mycology 52, no. 1 (2014): 36–45, https://doi.org/10.3109/13693786.2013.811552.