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Home / Equine Research Bank / Vet Sci / Preparation and Application of Clostridium perfringens Alpha...
Veterinary sciences2024; 11(8); 381; doi: 10.3390/vetsci11080381

Preparation and Application of Clostridium perfringens Alpha Toxin Nanobodies.

Abstract: All subtypes of () produce the alpha toxin (CPA), which can cause enteritis or enterotoxemia in lambs, cattle, pigs, and horses, as well as traumatic clostridial myonecrosis in humans and animals. CPA acts on cell membranes, ultimately leading to endocytosis and cell death. Therefore, the neutralization of CPA is crucial for the prevention and treatment of diseases caused by . In this study, utilizing CPA as an antigen, a nanobody (CPA-VHH) with a half-life of 2.9 h, an affinity constant (KD) of 0.9 nmol/L, and good stability below 60 °C was prepared from a natural nanobody library from alpacas. The biological activity analysis of CPA-VHH revealed its ability to effectively neutralize the phospholipase and hemolytic activity of CPA at a 15-fold ratio. In Vero cells, 9.8 μg/mL CPA-VHH neutralized the cytotoxicity of CPA at two times the half-maximal inhibitory concentration (IC). In a mouse model, 35.7 ng/g body weight (BW) of CPA-VHH neutralized 90% of the lethality caused by a 2× median lethal dose (LD) of CPA. It was found that CPA-VHH protected 80% of mice within 30 min at 2 × LD CPA, but this dropped below 50% after 2 h and to 0% after 4 h. Rescue trials indicated that using CPA-VHH within 30 min post-infection with 2 × LD CPA achieved an 80% rescue rate, which decreased to 10% after 2 h. Furthermore, CPA-VHH effectively mitigated the reduction in the expression levels of zonula occludens-1 (ZO-1), Occludin, and Claudin-1, while also attenuating the upregulation of the pro-inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-7 (IL-7), interleukin-8 (IL-8), tumor necrosis factor α (TNF-α), and interferon-γ (IFN-γ) induced by CPA infection. Overall, this study has identified a specific nanobody, CPA-VHH, that effectively neutralizes CPA toxins in vitro and in animal models, providing a new tool for inhibiting the pathogenicity resulting from these toxins and laying an important foundation for the development of new anti- toxin-related therapeutic products.
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Publication Date: 2024-08-19 PubMed ID: 39195835PubMed Central: PMC11360521DOI: 10.3390/vetsci11080381Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research involved the creation and testing of a specific nanobody, termed CPA-VHH. This nanobody was developed to effectively neutralize toxins produced by Clostridium perfringens. The study found that this nanobody could effectively neutralize the toxins, offering potential for future therapeutic uses.

Development of CPA-VHH Nanobody

  • The researchers used alpha toxin (CPA) from Clostridium perfringens as an antigen.
  • They developed a nanobody, called CPA-VHH, from a natural nanobody library obtained from alpacas.
  • CPA-VHH showcased good stability below 60 °C and had a half-life of 2.9 hours with an affinity constant of 0.9 nmol/L.

Biological Activity Analysis

  • The viability of CPA-VHH was confirmed by its ability to neutralize phospholipase and hemolytic activity of CPA.
  • In Vero cells, CPA-VHH neutralized the cytotoxicity of CPA at twice the half-maximal inhibitory concentration.
  • In a testing on mice, CPA-VHH neutralized 90% of the lethality caused by the median lethal dose of CPA.

Effective Use of CPA-VHH

  • CPA-VHH offered 80% protection within 30 minutes at twice the lethal dosage level, declining later.
  • It proved successful in rescue trials if used within 30 minutes post-exposure to the toxin.
  • CPA-VHH was also effective in downregulating pro-inflammatory cytokines induced by CPA infection.

Implications and Applications

  • This study highlights the potential use of nanobodies like CPA-VHH in neutralizing toxins from Clostridium perfringens.
  • These findings lay the foundation for future development of clostridium perfringens toxin-related therapeutic products.

Cite This Article

APA
Jia Q, Ren H, Zhang S, Yang H, Gao S, Fan R. (2024). Preparation and Application of Clostridium perfringens Alpha Toxin Nanobodies. Vet Sci, 11(8), 381. https://doi.org/10.3390/vetsci11080381

Publication

Veterinary sciences
ISSN: 2306-7381
NlmUniqueID: 101680127
Country: Switzerland
Language: English
Volume: 11
Issue: 8
PII: 381

Researcher Affiliations

Jia, Qiong
  • College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China.
Ren, Hongrui
  • College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China.
Zhang, Shuyin
  • College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China.
Yang, Haoyu
  • College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China.
Gao, Shuaipeng
  • College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China.
Fan, Ruiwen
  • College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China.

Conflict of Interest Statement

The authors declare no conflicts of interest.

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