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Updated: Jun 6, 2026
Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291
Published on: December 10, 2016
In vivo studies of Clostridium perfringens in mouse gas gangrene model
Nabonita Sengupta1, Syed Imteyaz Alam
1Biotechnology Division, Defence Research & Development Establishment, Gwalior, 474002, India. nabonita.s@gmail.com
Abstract:
Understanding the pathogenesis of infectious diseases requires comprehensive knowledge of the proteins expressed by the pathogen during in vivo growth in the host. Proteomics provides the tools for such analyses but the protocols required to purify sufficient quantities of the pathogen from the host organism are currently lacking. In this study, we have separated Clostridium perfringens, a highly virulent bacterium and potential BTW agent, from the peritoneal fluid of infected mice using Percoll density gradient centrifugation. The bacterium could be isolated in quantities sufficient to carry out meaningful proteomic comparisons with in vitro grown bacteria. Furthermore, the isolates were found to be virtually free from contaminating host proteins. Microscopy revealed major morphological changes under host conditions at different stages of infection. Profile of immunogenic proteins from in vivo- and TPYG-grown whole cell lysate using mouse anti-gangrene serum indicated over-expression of several proteins especially in the low molecular weight region. Expression of two virulence determinants, ornithine carbamoyl transferase (cOTC), and cystathionine beta-lyase (CBL), under in vivo conditions has also been studied. Two-dimensional gel analysis revealed a host induced proteome which was apparently different in comparison to in vitro grown cells. Detailed proteomic elucidation of differentially expressed proteins shown here is likely to provide valuable insight towards understanding the complexity of the adaptive response of C. perfringens to the host environment.
Insights
Researchers developed a new method to isolate Clostridium perfringens bacteria from mice, enabling proteomic analysis. This study reveals key differences in bacterial protein expression between in vivo and in vitro conditions, crucial for understanding infectious disease pathogenesis.
Area of Science:
- Microbiology
- Proteomics
- Infectious Disease Pathogenesis
Background:
- Understanding pathogen protein expression during host infection is vital for infectious disease research.
- Current proteomic analyses are limited by the lack of effective methods to purify pathogens from host organisms.
- Clostridium perfringens is a highly virulent bacterium and a potential bioterrorism agent.
Purpose of the Study:
- To develop and validate a protocol for isolating Clostridium perfringens from host environments for proteomic analysis.
- To compare the proteomes of Clostridium perfringens grown in vivo versus in vitro.
- To identify host-induced changes in bacterial protein expression and virulence factors.
Main Methods:
- Percoll density gradient centrifugation was used to isolate Clostridium perfringens from the peritoneal fluid of infected mice.
- Proteomic analysis, including two-dimensional gel electrophoresis, was performed on isolated bacteria.
- Immunogenic protein profiles were analyzed using mouse anti-gangrene serum.
- Expression of virulence determinants ornithine carbamoyl transferase (cOTC) and cystathionine beta-lyase (CBL) was studied.
Main Results:
- A method was established to purify Clostridium perfringens from host samples with minimal contamination.
- Significant morphological changes were observed in bacteria under host conditions.
- Proteomic analysis revealed differential expression of proteins, particularly in the low molecular weight range, between in vivo and in vitro grown bacteria.
- Host-induced proteome alterations were identified, suggesting an adaptive response.
Conclusions:
- The developed isolation protocol enables meaningful proteomic comparisons of Clostridium perfringens under host conditions.
- Proteomic analysis reveals significant differences in bacterial protein expression in vivo, highlighting the host's influence.
- This study provides valuable insights into the adaptive strategies of Clostridium perfringens during infection, aiding in understanding pathogenesis.
