An improved whole cell pertussis vaccine with reduced content of endotoxin

Human Vaccines & Immunotherapeutics. 2012 February; 9(2): 339–348

Waldely Oliveira Diasa, Arno A. J. van der Arkb, Maria Aparecida Sakauchic, Flávia Saldanha Kubruslya, Ana Fabíola R. O. Prestesa, Monamaris Marques Borgesd, Noemi Furuyamac, Denise S.P.Q. Hortond, Wagner Quintilioe, Marta Antoniazif, Betsy Kuipersb, Bernard A.M. van der Zeijstband Isaias Rawa

aCenter of Biotechnology, Instituto Butantan, SP Brazil

bResearch and Development, Netherlands Vaccine Institute (NVI)/ National Institute for Public Health and the Environment (RIVM)

cDivision of Technological Development and Production, Instituto Butantan, SP Brazil

dLaboratory of Bacteriology, Instituto Butantan, SP Brazil

eQuality Control Service, Instituto Butantan, SP Brazil

fLaboratory of Genetic, Instituto Butantan, SP Brazil



An improved whole cell pertussis vaccine (wP), designated as Plow, which is low in endotoxicity due to a chemical extraction of lipo-oligosaccharide (LOS) from the outer membrane, was evaluated for safety, immunogenicity and potency, comparatively to a traditional whole cell pertussis vaccine.

Current whole cell pertussis vaccines are effective but contain large quantities of endotoxin and consequently display local and systemic adverse reactions after administration. Endotoxin is highly inflammatory and contributes considerably to the reactogenicity as well as the potency of these vaccines. In contrast, acellular pertussis vaccines hardly contain endotoxin and are significantly less reactogenic, but their elevated costs limit their global use, especially in developing countries. In this paper, bulk products of Plow and a traditional whole cell vaccine, formulated as plain monocomponents or combined with diphtheria and tetanus toxoids (DTPlow or DTP, respectively) were compared by in vitro and in vivo assays. Chemical extraction of LOS resulted in a significant decrease in endotoxin content (20%) and a striking decline in endotoxin related toxicity (up to 97%), depending on the used in vitro or in vivo test. The LOS extraction did not affect the integrity of the product and, more importantly, did not affect the potency and/or stability of DTPlow. Moreover, hardly any differences in antibody and T-cell responses were observed. The development of Plow is a significant improvement regarding the endotoxicity of whole cell pertussis vaccines and therefore a promising and affordable alternative to currently available whole cell or acellular pertussis vaccines for developing countries.



Instituto Butantan (IB) in São Paulo, Brazil produces DTP vaccines for mass vaccination for more than 30 years. Currently, more than 90% of the Brazilian children are vaccinated at the age of 2–4-6 and 18 months of life, free of charge, representing about 250 million doses administered, annually.

Although rare, however, the risk of side effects related to wP remains a hindrance for its continued use worldwide. One of the most important factors that contribute to the reactogenicity of wP is the presence of lipo-oligosaccharide (LOS), the endotoxin from the bacterial outer membrane. The conventional wP detoxification procedures require a fastidious exposure of the cell suspension to formaldehyde, in order to obtain a decrease in toxicity.

Instituto Butantan developed an improved wP vaccine that is low in endotoxicity by a chemical treatment with a mixture of water and butanol, in a short overnight period, partially extracting the LOS from the B. pertussis outer membrane. Furthermore, the process also provides a by-product, a raw material for obtaining the monophosphoryl- lipid A (MPLA), which can be used as adjuvant for other vaccines, and was successfully coadministered with influenza vaccine (1, 2).

Our data strongly suggest a potential additional safety of whole cell pertussis vaccines, with possible cost-benefits gains in its production process.

Waldely de Oliveira Dias-1

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