PMID- 31811913
OWN - NLM
STAT- MEDLINE
DCOM- 20201214
LR  - 20201214
IS  - 1096-0279 (Electronic)
IS  - 1046-5928 (Linking)
VI  - 167
DP  - 2020 Mar
TI  - Immunization of rabbits with recombinant Clostridium perfringens alpha toxins 
      CPA-C and CTB-CPA-C in a bicistronic design expression system confers strong 
      protection against challenge.
PG  - 105550
LID - S1046-5928(19)30510-8 [pii]
LID - 10.1016/j.pep.2019.105550 [doi]
AB  - The Clostridium perfringens alpha toxin (CPA), encoded by the plc gene, is the 
      causative pathogen of gas gangrene, which is a lethal infection. In this study, 
      we used an E. coli system for the efficient production of recombinant proteins 
      and developed a bicistronic design (BCD) expression construct consisting of two 
      copies of the C-terminal (247-370) domain of the alpha toxin (CPA-C) in the first 
      cistron, followed by Cholera Toxin B (CTB) linked with another two copies of 
      CPA-C in the second cistron that is controlled by a single promoter. Rabbits were 
      immunized twice with purified proteins (rCPA-C rCTB-CPA-C) produced in the BCD 
      expression system, with an inactivated recombinant E. coli vaccine (RE), C. 
      perfringens formaldehyde-inactivated alpha toxoid (FA-CPA) and C. 
      perfringensl-lysine/formaldehyde alpha toxoid (LF-CPA) vaccines. Following the 
      second vaccination, 0.1 mL of pooled sera of the RE-vaccinated rabbits could 
      neutralize 12x mouse LD(100) (100% lethal dose) of CPA, while that of the rCPA-C 
      rCTB-CPA-C-vaccinated rabbits could neutralize 6x mouse LD(100) of CPA. Antibody 
      titers against CPA were also assessed by ELISA, reaching titers as high as 
      1:2048000 in the RE group; this was significantly higher compared to the C. 
      perfringens alpha toxoid vaccinated groups (FA-CPA and LF-CPA). Rabbits from all 
      vaccinated groups were completely protected from a 2x rabbit LD(100) of CPA 
      challenge. These results demonstrate that the recombinant proteins are able to 
      induce a strong immune responses, indicating that they may be potentially 
      utilized as targets for novel vaccines specifically against the C. perfringens 
      alpha toxin.
CI  - Copyright (c) 2019 Elsevier Inc. All rights reserved.
FAU - Peng, Xiaobing
AU  - Peng X
AD  - Department of Bacterial Biologics, China Institute of Veterinary Drug Control, 
      Beijing, China. Electronic address: 673303882@qq.com.
FAU - Peng, Guorui
AU  - Peng G
AD  - Department of Bacterial Biologics, China Institute of Veterinary Drug Control, 
      Beijing, China.
FAU - Li, Xuni
AU  - Li X
AD  - Department of Bacterial Biologics, China Institute of Veterinary Drug Control, 
      Beijing, China.
FAU - Feng, Lifang
AU  - Feng L
AD  - Good Clinical Practice Office, Beijing Zhonghai Biotech Co., Ltd, Beijing, China.
FAU - Dong, Lingying
AU  - Dong L
AD  - Department of Bacterial Biologics, China Institute of Veterinary Drug Control, 
      Beijing, China.
FAU - Jiang, Yuwen
AU  - Jiang Y
AD  - Department of Bacterial Biologics, China Institute of Veterinary Drug Control, 
      Beijing, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20191204
PL  - United States
TA  - Protein Expr Purif
JT  - Protein expression and purification
JID - 9101496
RN  - 0 (Antibodies, Bacterial)
RN  - 0 (Bacterial Toxins)
RN  - 0 (Bacterial Vaccines)
RN  - 0 (Calcium-Binding Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 9012-63-9 (Cholera Toxin)
RN  - EC 3.1.4.- (Type C Phospholipases)
RN  - EC 3.1.4.3 (alpha toxin, Clostridium perfringens)
SB  - IM
MH  - Animals
MH  - Antibodies, Bacterial/*blood
MH  - *Bacterial Toxins/biosynthesis/genetics/immunology/isolation & purification
MH  - Bacterial Vaccines
MH  - *Calcium-Binding Proteins/biosynthesis/genetics/immunology/isolation & 
      purification
MH  - Cholera Toxin/genetics
MH  - Cloning, Molecular
MH  - Clostridium perfringens/genetics/metabolism
MH  - Escherichia coli/genetics
MH  - Mice
MH  - Rabbits
MH  - *Recombinant Proteins/biosynthesis/genetics/immunology/isolation & purification
MH  - *Type C Phospholipases/biosynthesis/genetics/immunology/isolation & purification
MH  - Vaccination/methods
OTO - NOTNLM
OT  - Alpha toxin
OT  - Bicistron design system
OT  - Clostridum perfringens
OT  - ELISA
OT  - Recombinant vaccine
EDAT- 2019/12/08 06:00
MHDA- 2020/12/15 06:00
CRDT- 2019/12/08 06:00
PHST- 2019/09/18 00:00 [received]
PHST- 2019/12/03 00:00 [revised]
PHST- 2019/12/03 00:00 [accepted]
PHST- 2019/12/08 06:00 [pubmed]
PHST- 2020/12/15 06:00 [medline]
PHST- 2019/12/08 06:00 [entrez]
AID - S1046-5928(19)30510-8 [pii]
AID - 10.1016/j.pep.2019.105550 [doi]
PST - ppublish
SO  - Protein Expr Purif. 2020 Mar;167:105550. doi: 10.1016/j.pep.2019.105550. Epub 
      2019 Dec 4.