Recombinant diphtheria toxin derivatives: Perspectives of application

Romaniuk, S. I.; Dv, Kolibo; Комісаренко, С. В.

doi:10.1134/s106816201206012x

Cited by 11 publications

(4 citation statements)

References 140 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…At the LB/kanamycin media, the transformed E. coli DH5α and E. coli BL21 were positive for growth. These findings have shown that the progress of the genetic transformation and gene transfer experiences under research has been observed and is also compatible with several previous studies, such as Mohammadi et al [ 21 ], Nascimento et al [ 22 ], and Romaniuk et al [ 28 ].…”

Section: Discussionsupporting

confidence: 93%

Expression of full and fragment-B of diphtheria toxin genes inEscherichia colifor generating of recombinant diphtheria vaccines

Abulmagd

Khattab

Zedan

2022

Clin Exp Vaccine Res

View full text Add to dashboard Cite

Purpose In the present study, whole diphtheria toxin (dt) and fragment B (dtb) genes from Corynebacterium diphtheriae Park William were cloned into Escherichia coli , the purified expressed proteins were evaluated for ultimately using as a candidate vaccine. Materials and Methods The dt and dtb genes were isolated from bacterial strain ATCC (American Type Culture Collection) no. 13812. Plasmid pET29a+ was extracted by DNA-spin TM plasmid purification kit where genes were inserted using BamHI and HindIII-HF. Cloned pET29a+dt and pET29a+dtb plasmids were transformed into E. coli BL21(DE3)PlysS as expression host. The identity of the sequences was validated by blasting the sequence (BLASTn) against all the reported nucleotide sequences in the NCBI (National Center for Biotechnology Information) GenBank. Production of proteins in high yield by different types and parameters of fermentation to determine optimal conditions. Lastly, the purified concentrated rdtx and rdtb were injected to BALB/c mice and antibody titers were detected. Results The genetic transformation of E. coli DH5α and E. coli BL21 with the pET-29a(+) carrying the dt and dtb genes was confirmed by colony polymerase chain reaction assay and were positive to grow on Luria-Bertani/kanamycin medium. The open reading frame of dt and dtb sequences consisted of 1,600 bp and 1,000 bp, were found to be 100% identical to dt and dtb sequence of C. diphtheriae (accession number KX702999.1 and KX702993.1) respectively. The optimal condition for high cell density is fed-batch fermentation production to express the rdtx and rdtb at 280 and 240 Lf/mL, dissolved oxygen was about 24% and 22% and the dry cell weight of bacteria was 2.41 g/L and 2.18 g/L, respectively. Conclusion This study concluded with success in preparing genetically modified two strains for the production of a diphtheria vaccine, and to reach ideal production conditions to achieve the highest productivity.

show abstract

Section: Discussionsupporting

confidence: 93%

Expression of full and fragment-B of diphtheria toxin genes inEscherichia colifor generating of recombinant diphtheria vaccines

Abulmagd

Khattab

Zedan

2022

Clin Exp Vaccine Res

View full text Add to dashboard Cite

show abstract

“…However, DTT is better than DTB with regard to antigen load. DTT possesses only one B cell epitope whereas DTB contains several highly immunogenic B cell epitopes in the R domain [ 42 ]. The B cell epitope of DT has been shown to suppress the immune response of certain conjugated haptens by competing DT primed helper T cells [ 43 ].…”

Section: Discussionmentioning

confidence: 99%

A Rationally Designed TNF-α Epitope-Scaffold Immunogen Induces Sustained Antibody Response and Alleviates Collagen-Induced Arthritis in Mice

Zhang

Wang

et al. 2016

PLoS ONE

View full text Add to dashboard Cite

The TNF-α biological inhibitors have significantly improved the clinical outcomes of many autoimmune diseases, in particular rheumatoid arthritis. However, the practical uses are limited due to high costs and the risk of anti-drug antibody responses. Attempts to develop anti-TNF-α vaccines have generated encouraging data in animal models, however, data from clinical trials have not met expectations. In present study, we designed a TNF-α epitope-scaffold immunogen DTNF7 using the transmembrane domain of diphtheria toxin, named DTT as a scaffold. Molecular dynamics simulation shows that the grafted TNF-α epitope is entirely surface-exposed and presented in a native-like conformation while the rigid helical structure of DTT is minimally perturbed, thereby rendering the immunogen highly stable. Immunization of mice with alum formulated DTNF7 induced humoral responses against native TNF-α, and the antibody titer was sustained for more than 6 months, which supports a role of the universal CD4 T cell epitopes of DTT in breaking self-immune tolerance. In a mouse model of rheumatoid arthritis, DTNF7-alum vaccination markedly delayed the onset of collagen-induced arthritis, and reduced incidence as well as clinical score. DTT is presumed safe as an epitope carrier because a catalytic inactive mutant of diphtheria toxin, CRM197 has good clinical safety records as an active vaccine component. Taken all together, we show that DTT-based epitope vaccine is a promising strategy for prevention and treatment of autoimmune diseases.

show abstract

“…Non-toxic DT mutants (cross-reacting materials) have already been applied in animal models to induce effective anti-diphtheria immune response ( 14 ). For example, in our previous study non-toxic recombinant subunit B of DT has been proposed as a safe and effective antigen with a good potential to elicit protective immune response in immunized guinea pigs ( 15 ).…”

Section: Discussionmentioning

confidence: 99%

Cellobiose-coated poly (lactide-co-glycolide) particles loaded with diphtheria toxoid for per os immunization

Chudina

Labyntsev²,

Manoilov³

et al. 2015

Croat Med J

View full text Add to dashboard Cite

AimTo evaluate the dose-dependent immunogenic properties of poly (lactide-co-glycolide) (PLGA) particles coated with cellobiose as antigen carriers for oral immunization.MethodsTwo types of PLGA-cellobiose particles (PLGA-cellobiose-1, ~ 0.8 μm and PLGA-cellobiose-2, ~ 1.2 μm) containing non-toxic recombinant subunit B (SbB) of diphtheria toxin fused with enhanced green fluorescent protein were characterized in vitro for their size, shape, antigen loading, and ability to induce phagocytosis. Different doses of antigen, immobilized on the particles (2.5 μg, 25 μg, 250 μg, and 2500 μg per 1 kg of body weight), were administered per os 3 times with intervals of 2 weeks to BALB/c female mice. The antigen-specific IgG and IgA antibodies were estimated in serum by ELISA.ResultsAfter the first immunization, increase in concentration of blood antitoxic antibodies was detected. Antigen dose 250 μg/kg was the most immunogenic for IgG antibodies induction for both types of PLGA-cellobiose particles. Antigen doses 25 μg/kg and 2.5 μg/kg were the most immunogenic for IgA antibodies induction by PLGA-cellobiose 1 and 2 particles, respectively. The second and the third treatment had no significant effect on the immune response or even reduced it, which could be explained by immune tolerance induction by the antigens delivered per os.ConclusionOur results suggest that the correct dose of PLGA-cellobiose particles loaded with antigen could significantly increase the humoral immune response against the introduced antigen already after the first immunization. Thus, PLGA particles can be considered as a potent component of oral vaccines.

show abstract

Recombinant diphtheria toxin derivatives: Perspectives of application

Cited by 11 publications

References 140 publications

Expression of full and fragment-B of diphtheria toxin genes inEscherichia colifor generating of recombinant diphtheria vaccines

Expression of full and fragment-B of diphtheria toxin genes inEscherichia colifor generating of recombinant diphtheria vaccines

A Rationally Designed TNF-α Epitope-Scaffold Immunogen Induces Sustained Antibody Response and Alleviates Collagen-Induced Arthritis in Mice

Cellobiose-coated poly (lactide-co-glycolide) particles loaded with diphtheria toxoid for per os immunization

Contact Info

Product

Resources

About