Standardization and validation of Vero cell assay for potency estimation of diphtheria antitoxin serum

Kumar, Sunil; Kanwar, Sarika; Bansal, Vivek; Sehgal, Rakesh

doi:10.1016/j.biologicals.2009.05.002

Cited by 10 publications

(7 citation statements)

References 2 publications

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“…The chemical substances tested include carbamazepine [79]; triclosan [80]; lead nitrate [81]; pentachlorophenol and rotenone [82], where the Vero cell line revealed to be one of the most sensitive model used in these studies. These cells have also been validated as a cellular model for other microbial toxins such as diphtheria toxin, a polypeptide with 535 a.a. [83] and Shiga-like toxins, a protein of enterohemorrhagic Escherichia coli [84]. The sensitivity of Vero cell line to another cyanobacterial toxin, cylindrospermopsin, was also reported although, such as MCs, the mechanism of toxin uptake by this cell line remains to clarify [85,86].…”

Section: Is Vero-e6 Cell Line a Suitable Model To Study Toxicologicalmentioning

confidence: 99%

The Kidney Vero-E6 Cell Line: A Suitable Model to Study the Toxicity of Microcystins

Menezes¹,

Valério²,

Dias³

2013

New Insights Into Toxicity and Drug Testing

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Section: Is Vero-e6 Cell Line a Suitable Model To Study Toxicologicalmentioning

confidence: 99%

The Kidney Vero-E6 Cell Line: A Suitable Model to Study the Toxicity of Microcystins

Menezes¹,

Valério²,

Dias³

2013

New Insights Into Toxicity and Drug Testing

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“…breast (MDA-MB-231 and MCF-7), and liver (HepG2) have been used to investigate the anticancer potential of Phoenix dactylifera seed extract (PDSE). African green monkey normal kidney epithelial cell line (Vero) has been widely used in various toxicological, virology and pharmacological research domains, particularly as a sensitive model for toxicity assays of compounds of various nature, either chemical or microbial toxins [ 19 – 22 ]. Thus, to test the toxicity of PDSE, Vero cell line was used as a control to mimic normal mammalian cells.…”

Section: Introductionmentioning

confidence: 99%

Anticancer potential of Phoenix dactylifera L. seed extract in human cancer cells and pro-apoptotic effects mediated through caspase-3 dependent pathway in human breast cancer MDA-MB-231 cells: an in vitro and in silico investigation

Khan

Singh

Siddiqui

et al. 2022

BMC Complement Med Ther

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Background Phoenix dactylifera L. has a diverse set of pharmacological properties due to its distinct phytochemical profile. The purpose of this study was to investigate the anticancer potential of Phoenix dactylifera seed extract (PDSE) in human breast cancer MDA-MB-231 and MCF-7 cells, as well as liver cancer HepG2 cells, and to investigate the anticancer efficacy in triple-negative MDA-MB-231 cells, followed by in silico validation of the molecular interaction between active components of PDSE and caspase-3, an apoptosis executioner protein . Methods In this study, human cancer cell lines were cultured and subsequently treated with 10 to 100 μg/mL of PDSE. MTT test was performed to determine the cell viability, MMP was measured using fluorescent probe JC-1, nuclear condensation was determined by Hoechst 33258 dye, Annexin V-FITC & PI staining and cell cycle analysis were evaluated through flow cytometer, and apoptotic markers were detected using western blotting. The bioactive agents in PDSE were identified using high-performance liquid chromatography (HPLC) analysis. The binding affinity was validated using molecular docking tools AutoDock Vina and iGEMDOCK v2.1. Results Cell viability data indicated that PDSE inhibited cell proliferation in both breast cancer cells and liver cancer cells. MDA-MB-231 cells showed maximum growth inhibition with an IC50 value of 85.86 μg/mL for PDSE. However, PDSE did not show any significant toxicity against the normal Vero cell line. PDSE induced MMP loss and formation of apoptotic bodies, enhanced late apoptosis at high doses and arrested cells in the S phase of cell cycle. PDSE activated the enzymatic activity of cleaved caspase-3 and caused the cleavage of poly-ADB ribose polymerase (PARP) protein. PDSE upregulated pro-apoptotic Bax protein markedly but no significant effect on tumor suppressor protein p53, while it downregulated the anti-apoptotic Bcl-2 protein expression. HPLC analysis showed the presence of rutin and quercetin bioactive flavonols in ethanolic extract of PDS. Interestingly, both active components revealed a strong binding interaction with amino acid residues of caspase-3 (PDB ID: 2XYP; Hetero 4-mer - A2B2) protein. Conclusion PDS could serve as a potential medicinal source for apoptotic cell death in human breast cancer cells and, thus, could be used as a promising and crucial candidate in anticancer drug development. This study warrants further in vivo research, followed by clinical investigation.

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“…Response to the vaccination can be measured by in vivo or in vitro methods. The in vivo method is the gold standard, which measures the ability of antibodies to neutralize diphtheria toxin in experimental animals [21], but this method is very expensive. In vitro measurement methods include toxin binding inhibition assay (ToBI), ELISA, haemagglutination assay, and Vero cell testing.…”

Section: Methodsmentioning

confidence: 99%

“…ToBI and ELISA measure toxin-specific antibodies in total but do not distinguish between neutralizing and non-neutralizing antibodies. Vero cell assay (VCA) has a high degree of correlation with the in vivo biological assay and is therefore recommended by WHO as an alternative to the in vivo assay method [21,22]. We use the VCA method in this study.…”

Section: Methodsmentioning

confidence: 99%

Post Vaccination Titers of Serum Anti-Diphtheria in Indonesian Young and Middle-Aged Adults

Soegiarto¹,

Novritasari²,

Baskoro³

et al. 2021

Preprint

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Objective: This study aimed to evaluate the antibody responses in two adult age groups after diphtheria vaccination. Study Design: An observational analytic study was carried out to determine the difference in serum titer of anti-diphtheria antibody. Methods: Serum antibody titers were measured just before and 3 months after injection of Diphtheria toxoid vaccine. Vaccine was given to two adult age groups of health care personnel in hospital: the young (< 40 years) and the middle-aged (≥ 40 years). Data were analyzed using the Mann-Whitney test (p < 0.05). Results: Significant increase in serum anti-diphtheria antibody titers were recorded after vaccination in both age group (p < 0.001 in young adult and p = 0.001 in middle-aged adult, respectively). There were no substantial differences between the two groups in terms of antibody titer before vaccination (p = 0.741), 3 months after vaccination (p = 0.317) and in the increase of antibody titer (p = 0.479). Conclusions: This study showed that there was no significant difference in the increase of anti-diphtheria antibody titers between the two age groups, proving that both young and middle-aged adults had an equal immune response to a given diphtheria vaccine.

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Standardization and validation of Vero cell assay for potency estimation of diphtheria antitoxin serum

Cited by 10 publications

References 2 publications

The Kidney Vero-E6 Cell Line: A Suitable Model to Study the Toxicity of Microcystins

The Kidney Vero-E6 Cell Line: A Suitable Model to Study the Toxicity of Microcystins

Anticancer potential of Phoenix dactylifera L. seed extract in human cancer cells and pro-apoptotic effects mediated through caspase-3 dependent pathway in human breast cancer MDA-MB-231 cells: an in vitro and in silico investigation

Post Vaccination Titers of Serum Anti-Diphtheria in Indonesian Young and Middle-Aged Adults

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