The quantitative analysis of the mechanism involved in pertussis toxin-mediated cell clustering and its implications in the in vitro quality control of diphtheria tetanus and whole cell pertussis vaccines

Autor(es)

da Silva Zamith, Helena Pereira ^[1] ; Godinho, Rosely Oliveira ^[2] ; da Costa Junior, Valter Luiz ^[2] ; Corrado, Alexandre Pinto ^[3]

Afiliação do(s) autor(es)

^[1] Fiocruz MS, Fundacao Oswaldo Cruz, Inst Nacl Controle Qualidade Saude, Dept Farmacol & Toxicol, INCQS, Ave Brasil 4365, BR-21040900 Rio De Janeiro, RJ - Brazil

^[2] Univ Fed Sao Paulo EPM UNIFESP, Escola Paulista Med, Dept Farmacol, Rua Tres Maio 100, BR-04044020 Sao Paulo, SP - Brazil

^[3] Univ Sao Paulo FMRP USP, Fac Med Ribeirao Preto, Dept Farmacol, Av Bandeirantes 3900, BR-14049900 Ribeirao Preto, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: TOXICOLOGY IN VITRO; v. 70, FEB 2021.
• Citações Web of Science: 0

Resumo

Some of the adverse side-effects such as leukocytosis, hyperinsulinemia, hypoglycemia and sensitization to histamine, caused by diphtheria, tetanus and whole cell pertussis (DTwP) vaccines are related to the presence of non-inactivated pertussis toxin (PTx) residues (NiPTxR). The CHO cell clustering assay is an in vitro assay to measure NiPTxR in DTwP vaccines based on the ability of active PTx to cause cellular clustering. To study the biochemical mechanism involved in the clustering effect in CHO cells induced by PTx and by two DTwP vaccines, the levels of total cyclic cAMP were measured and compared to those obtained after treatment with cholera toxin (CTx) able to induce CHO cells elongation instead of cell clustering. Our results showed an increment of cAMP levels by CTx and total cell elongation in CHO cells. However, changes in cAMP levels were not associated with the total clustering induced by PTx or by DTwP vaccines. The high correlation seen between the levels of NiPTxR in the DTwP vaccines determined by the in vivo lethal histamine sensitization (HIST) assay and the in vitro CHO cell clustering assay indicated that the latter could be a suitable alternative test to HIST assay for the toxicological approval and release of batches of DTwP vaccines in their final formulation for human use in accordance with the application of the 3R's principle. (AU)

• Processo FAPESP: 18/21381-6 - Validação da "Via Extracelular AMPc-Adenosina" como novo alvo terapêutico para o tratamento de doenças respiratórias crônicas
• Beneficiário: Rosely Oliveira Godinho
• Modalidade de apoio: Auxílio à Pesquisa - Regular