The Transcriptional Regulatory Network of Corynebacterium pseudotuberculosis

Parise, Doglas; Parise, Mariana Teixeira Dornelles; Gomide, Anne Cybelle Pinto; Aburjaile, Flávia Figueira; Kato, Rodrigo Bentes; Salgado-Albarrán, Marisol; Tauch, Andreas; Azevedo, Vasco; Baumbach, Jan

doi:10.3390/microorganisms9020415

“…For example, Corynebacterium pseudotuberculosis infection is one such disease that leads to disseminated granulomas in ruminants. This infection is highly contagious, and it can result in serious economic losses caused by reduced production efficiency and carcass disposal, making it an extremely serious infection in ruminants ( 33 ). The immunological diagnostic method established in this study is expected to be used to differentiate R. equi infection from similar diseases.…”

Section: Discussionmentioning

confidence: 99%

Development of monoclonal antibodies against Rhodococcus equi virulence-associated protein N and their application to pathological diagnosis

Suzuki,

Takai,

Morizane

et al. 2023

Microbiol Spectr

2

0

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Since 2015, virulence-associated protein N ( vapN )-positive Rhodococcus equi has been isolated from pyogenic lesions in ruminants in multiple countries, including Japan, supporting the widespread distribution of this pathogen in ruminants, and suggesting that VapN is responsible for the pathogenicity of this infection. However, no immunological diagnostic method has been established for this infection. In this study, we attempted to produce anti-VapN monoclonal antibodies and apply them to immunostaining methods. Mice were immunized intraperitoneally or subcutaneously with 5 µg of recombinant VapN (rVapN). After cell fusion and cloning by limiting dilution, we generated three anti-VapN antibody-producing hybridomas (4H4, 5C3, and 5G10). Next, VapN-based immunostaining was performed using a mouse model inoculated with rVapN, the JCM94-3 (VapN-producing) strain, or the JID03-27 (low-producing) strain. Positive reactions were observed in necrotic lesions in the liver and spleen, especially in the rVapN-treated and JCM94-3-treated groups. Immunostaining of goats ( n = 2) and cattle ( n = 3) presumed to be infected with vapN -positive R. equi was also positive in organs carrying necrotic lesions including the lungs, lymph nodes, abomasum, and ribs. The reactivity of immunostaining differed among the antibodies, with the 4H4 antibody exhibiting the best reactivity and allowing detection at low concentrations. However, the reactivity of the other two antibodies was improved by antigen retrieval treatment. Several known diseases cause necrotizing granuloma formation, and the immunological diagnostic method established in this study can be used to distinguish R. equi infection from similar diseases or identify infection in previously missed cases. IMPORTANCE Rhodococcus equi can cause infection in ruminants, and its pathogenicity is suggested to be associated with VapN. Despite its wide distribution, no immunological diagnostic method has been developed for VapN-producing R. equi . Against this background, we attempted to develop monoclonal antibodies targeting VapN and assess their application in immunostaining. In the study, mice were immunized with recombinant VapN, and cell fusion and cloning by limiting dilution permitted the generation of three antibody-producing hybridomas. The utility of the antibodies produced from the hybridomas in immunostaining was demonstrated using an infected mouse model, and the antibodies were further applied to previously reported cases of R. equi infection in goats and cattle. Although the 4H4 antibody induced the strongest reactions, the reactivity of two other antibodies was improved by antigen retrieval. Our monoclonal antibodies will be utilized to support the definitive diagnosis of suspected R. equi infection, including cases that were previously missed.

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“…glutamicum for economic reasons, as reviewed in [9], a recent report described that a ribonuclease in C. diphtheriae , RNase J, controls mRNA metabolism and therefore influences bacterial physiology (division, growth, cellular shape) and virulence [10], an example that this kind of study on pathogenic Corynebacterium generates valuable information about the pathogenesis. Members of the genus Corynebacterium are Gram-stain-positive, facultatively anaerobic and rod-shaped, with ranging G+C contents of 46–74 mol% [11].…”

Section: Full-textmentioning

confidence: 99%

Corynebacterium lizhenjunii sp. nov., isolated from the respiratory tract of Marmota himalayana, and Corynebacterium qintianiae sp. nov., isolated from the lung tissue of Pseudois nayaur

Zhou

¹

,

Xu

²

,

Guo

³

et al. 2021

International Journal of Systematic and Evolutionary Microbiology

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Four Gram-stain-positive, non-motile and asporous bacilli (strains ZJ-599T, ZJ-621, MC1420T and MC1482), isolated from animal tissue and environmental samples collected on the Qinghai-Tibet Plateau, PR China, were taxonomically characterized. Based on the results of 16S rRNA gene sequence analyses, the closest relatives of strains ZJ-599T and ZJ-621 were Corynebacterium endometrii LMM-1653T (97.5 %), Corynebacterium phocae M408/89/1T (96.5 %) and Corynebacterium flavescens OJ8T (96.3 %), whereas strains MC1420T and MC1482 were closest to Corynebacterium sanguinis CCUG 58655T (98.9 %), Corynebacterium mycetoides DSM 20632T (98.4 %) and Corynebacterium lipophiloflavum DSM 44291T (97.9 %). The results of rpoB gene sequence similarity analysis indicated that C. phocae M408/89/1T and C. sanguinis CCUG 58655T were closest to strains ZJ-599T/ZJ-621 (83.5 %) and MC1420T/MC1482 (91.8 %), respectively. The two novel type strains shared a similarity of 95.2 % in 16S rRNA and 81.3 % in rpoB gene sequences. The TAP-PCR DNA fingerprint and MALDI-TOF MS spectrum patterns clearly differentiated the novel isolates within and between each pair of strains. Strain ZJ-599T had 21.9–22.4 % digital DNA–DNA hybridization (dDDH) scores with C. endometrii LMM-1653T, C. phocae M408/89/1T and C. flavescens OJ8T, and 72.3–72.9 % of average nucleotide identity (ANI) with them. Similarly, strain MC1420T had 22.9–23.7 % dDDH values with C. sanguinis CCUG 58655T, C. mycetoides DSM 20632T and C. lipophiloflavum DSM 44291T, and 80.4–81.3 % ANI scores with them. Strain ZJ-599T had a 23.1 % dDDH value and 70.5 % ANI score with strain MC1420T, both below the corresponding thresholds for species delineation. Strains ZJ-599T and MC1420T both contain mycolic acids and have MK-8(H2) and MK-9(H2) as the predominant respiratory quinones, meso-diaminopimelic acid as the diagnostic diamino acid, and C18 : 1 ω9c as the main fatty acid. C17 : 1 ω8c and C15 : 1 ω8c were predominant in strain ZJ-599T in contrast to C17 : 1 ω7c being predominant in strain MC1420T. The main polar lipids in strain ZJ-599T were diphosphatidylglycerol, phosphatidylinositol and one unidentified glycolipid, while strain MC1420T had diphosphatidylglycerol, phosphatidylglycerol and one unidentified lipid as the major components. Since the two pairs of novel strains (ZJ-599T/ZJ-621, MC1420T/MC1482) distinctly differ from each other and from their nearest relatives, two novel species of the genus Corynebacterium are proposed, namely Corynebacterium lizhenjunii (type strain ZJ-599T=GDMCC 1.1779T=JCM 34341T) and Corynebacterium qintianiae (type strain MC1420T=GDMCC 1.1783T=JCM 34340T), respectively.

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“…In addition, pharmacotherapy for clinical patients is not satisfactory and recurrent, which poses difficulties in controlling and eradicating the disease ( 6 ). This further caused serious economic losses to livestock production ( 7 ). Consequently, there is an urgent need to develop novel preventive and control strategies to curb the onset and progression of diseases caused by C. p .…”

Section: Introductionmentioning

confidence: 99%

Lactobacillus acidophilus protects against Corynebacterium pseudotuberculosis infection by regulating the autophagy of macrophages and maintaining gut microbiota homeostasis in C57BL/6 mice

Li,

Jiang,

Cui

et al. 2024

mSystems

0

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Corynebacterium pseudotuberculosis ( C. p ), a facultative intracellular bacterium, is an important zoonotic pathogen that causes abscesses and pyogenic granulomas. The relationship between gut microbiota and host health or diseases has received increasing attention. However, the role of gut microbiota in the process of C. p infection is still unclear. In this study, we established a C. p infection model in C57BL/6 mice and examined the impact of preemptive oral administration Lactobacillus acidophilus ( L. acidophilus ) on infection. Our findings revealed that C. p infection led to pronounced pathological alterations in the liver and kidneys, characterized by abscess formation, intense inflammatory responses, and bacterial overload. Remarkably, these deleterious effects were greatly relieved by oral administration of L. acidophilus before infection with C. p . Additionally, we further found that during C. p infection, peritoneal macrophages (PMs) of mice orally administered with L. acidophilus accumulated more rapidly at sites of infection. Furthermore, our results showed that PMs from mice with oral L. acidophilus administration showed a stronger C. p clearance effect, and this was mediated by high expression of LC3-II protein. Meanwhile, oral administration of L. acidophilus protected the gut microbiota disorder in C57BL/6 mice caused by C. p infection. In summary, our study demonstrates that oral administration of L. acidophilus confers effective protection against C. p infection in C57BL/6 mice by modulating macrophage autophagy, thereby augmenting bacterial clearance and preserving gut microbiota and function stability. These findings position L. acidophilus as a viable probiotic candidate for the clinical prevention of C. p infection. IMPORTANCE Corynebacterium pseudotuberculosis (C. p ) is known to induce a range of chronic diseases in both animals and humans. Currently, clinical treatment for C. p infection mainly relies on antibiotic therapy or surgical intervention. However, excessive use of antibiotics may increase the risk of drug-resistant strains, and the effectiveness of treatment remains unsatisfactory. Furthermore, surgical procedures do not completely eradicate pathogens and can easily cause environmental pollution. Probiotic interventions are receiving increasing attention for improving the body’s immune system and maintaining health. In this study, we established a C. p infection model in C57BL/6 mice to explore the impact of Lactobacillus acidophilus during C. p infection. Our results showed that L. acidophilus effectively protected against C. p infection by regulating the autophagy of macrophages and maintaining intestinal microbiota homeostasis. This study may provide a new strategy for the prevention of C. p infection.

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The Transcriptional Regulatory Network of Corynebacterium pseudotuberculosis

Cited by 5 publications

References 143 publications

Development of monoclonal antibodies against Rhodococcus equi virulence-associated protein N and their application to pathological diagnosis

Development of monoclonal antibodies against Rhodococcus equi virulence-associated protein N and their application to pathological diagnosis

Corynebacterium lizhenjunii sp. nov., isolated from the respiratory tract of Marmota himalayana, and Corynebacterium qintianiae sp. nov., isolated from the lung tissue of Pseudois nayaur

Lactobacillus acidophilus protects against Corynebacterium pseudotuberculosis infection by regulating the autophagy of macrophages and maintaining gut microbiota homeostasis in C57BL/6 mice

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