Mem Inst Oswaldo Cruz, Rio de Janeiro, 112(12) December 2017
Original Article

Parenteral adjuvant potential of recombinant B subunit of Escherichia coli heat-labile enterotoxin

Carlos Eduardo Pouey da Cunha+, Clóvis Moreira Junior, Andréa da Silva Ramos Rocha, Paula Fonseca Finger, Carolina Georg Magalhães, Marcos Roberto Alves Ferreira, Odir Antônio Dellagostin, Ângela Nunes Moreira, Fabricio Rochedo Conceição

Universidade Federal de Pelotas, Centro de Desenvolvimento Tecnológico, Biotecnologia, Pelotas, RS, Brasil

DOI: 10.1590/0074-02760170133
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ABSTRACT

BACKGROUND The B subunit of Escherichia coli heat-labile enterotoxin (LTB) 1 is a potent mucosal immune adjuvant. However, there is little information about LTBu2019s potential as a parenteral adjuvant.

OBJECTIVES We aimed at evaluating and better understanding rLTBu2019s potential as a parenteral adjuvant using the fused R1 repeat of Mycoplasma hyopneumoniae P97 adhesin as an antigen to characterise the humoral immune response induced by this construct and comparing it to that generated when aluminium hydroxide is used as adjuvant instead.

METHODS BALB/c mice were immunised intraperitoneally with either rLTBR1 or recombinant R1 adsorbed onto aluminium hydroxide. The levels of systemic anti-rR1 antibodies (total Ig, IgG1, IgG2a, and IgA) were assessed by enzyme-linked immunosorbent assay (ELISA). The ratio of IgG1 and IgG2a was used to characterise a Th1, Th2, or mixed Th1/Th2 immune response.

FINDINGS Western blot confirmed rR1, either alone or fused to LTB, remained antigenic; anti-cholera toxin ELISA confirmed that LTB retained its activity when expressed in a heterologous system. Mice immunised with the rLTBR1 fusion protein produced approximately twice as much anti-rR1 immunoglobulins as mice vaccinated with rR1 adsorbed onto aluminium hydroxide. Animals vaccinated with either rLTBR1 or rR1 adsorbed onto aluminium hydroxide presented a mixed Th1/Th2 immune response. We speculate this might be a result of rR1 immune modulation rather than adjuvant modulation. Mice immunised with rLTBR1 produced approximately 1.5-fold more serum IgA than animals immunised with rR1 and aluminium hydroxide.

MAIN CONCLUSIONS The results suggest that rLTB is a more powerful parenteral adjuvant than aluminium hydroxide when administered intraperitoneally as it induced higher antibody titres. Therefore, we recommend that rLTB be considered an alternative adjuvant, even if different administration routes are employed.

Financial support: CAPES, CNPq, FAPERGS
CEPC and CMJ contributed equally to this work.
+ Corresponding author: cpouey@gmail.com
Received 4 April 2017
Accepted 29 July 2017

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