Immune response in hamsters immunised with a recombinant fragment of LigA from <i>Leptospira interrogans</i>, associated with carrier molecules

Immunisation with the C-terminal region of leptospiral immunoglobulin-like A protein (LigANI) has shown promising results against leptospirosis. We evaluated the humoral immune response and protection induced by LigANI associated with carboxyl multi-walled carbon nanotubes (COOH-MWCNTs), CpG oligodeoxynucleotides (CpG ODNs), or Alhydrogel. Animals immunised with CpG ODNs were unable to develop a humoral immune response, whereas immunisation with LigANI and COOH-MWCNTs produced a high level of IgG antibodies, similar to that with LigANI and Alhydrogel, but it was not protective. The use of carbon nanotubes as an adjuvant in subunit vaccines against leptospirosis is a novel approach for improving specific IgG production.

Leptospira; LigANI; carrier molecules

Authorship

Thaís L Oliveira

Universidade Federal de Pelotas, Centro de Desenvolvimento Tecnológico, Programa de Pós-Graduação em Biotecnologia, Núcleo de Biotecnologia, Pelotas, RS, BrasilUniversidade Federal de PelotasBrasilPelotas, RS, BrasilUniversidade Federal de Pelotas, Centro de Desenvolvimento Tecnológico, Programa de Pós-Graduação em Biotecnologia, Núcleo de Biotecnologia, Pelotas, RS, Brasil

Kátia L Bacelo

Rodrigo A Schuch

Fabiana K Seixas

Tiago Collares

Oscar ED Rodrigues

Universidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, BrasilUniversidade Federal de Santa MariaBrasilSanta Maria, RS, BrasilUniversidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brasil

Josimar Vargas

Rafaella O do Nascimento

Montreal University, Chemistry Department, Montreal, QC, CanadaMontreal UniversityCanadaMontreal, QC, CanadaMontreal University, Chemistry Department, Montreal, QC, Canada

Odir A Dellagostin

Daiane D Hartwig ⁺+ Corresponding author: daianehartwig@gmail.com

Universidade Federal de Pelotas, Instituto de Biologia, Departamento de Microbiologia e Parasitologia, Pelotas, RS, BrasilUniversidade Federal de PelotasBrasilPelotas, RS, BrasilUniversidade Federal de Pelotas, Instituto de Biologia, Departamento de Microbiologia e Parasitologia, Pelotas, RS, Brasil

+ Corresponding author: daianehartwig@gmail.com

TLO and KLB contributed equally to this work.

AJAM and OAD are inventors on a patent submission entitled: LigA and LigB proteins [Leptospiral Ig-like (Lig) domains] for vaccination and diagnosis (Patent nº BRPI0505529 and WO 2007070996).

SCIMAGO INSTITUTIONS RANKINGS

Montreal University, Chemistry Department, Montreal, QC, CanadaMontreal UniversityCanadaMontreal, QC, CanadaMontreal University, Chemistry Department, Montreal, QC, Canada

Figures

Figures (4)

Thumbnail

Fig. 1
: cytotoxic effect of carbon nanotubes in CHO-K1 cells. The cytotoxicity of carbon nanotubes was assessed using MTT assay. The inhibition rate was expressed as the optical density ratio of treated cells compared to the negative control cells (only medium). The positive control cells were treated with 1% dimethyl sulfoxide. The data are expressed as mean ± standard deviation of three independent experiments. Asterisks indicate significant differences (p < 0.001) compared to the positive control.

Thumbnail

Fig. 2
: induction of IgG antibody response in hamsters immunised with different rLigANI vaccine preparations evaluated by ELISA. Values presented are means ± standard deviation of two independent experiments. Asterisks indicate significant differences (p < 0.05) compared to rLigANI-PBS group [rLigANI- phosphate-buffered saline (PBS), recombinant non-identical carboxy-terminus portion of LigA protein in PBS].

Thumbnail

Fig. 3
: isotyping of anti-rLigANI IgG subclasses. The data represent the difference between mean absorbance of hamster sera collected at day 28 and mean absorbance of hamster pre-immune sera at day 0. Results are expressed as mean absorbance ± standard deviation of pooled serum samples assayed in triplicates in two independent experiments. Asterisks indicate significant differences (p < 0.05) compared to rLigANI- phosphate-buffered saline (PBS) group (*) or compared to rLigANI-Alhydrogel group (**) (rLigANI-PBS, recombinant non-identical carboxy-terminus portion of LigA protein in PBS; rLigANI-Alhydrogel, rLigANI in 15% Alhydrogel).

Thumbnail

Fig. 4
: survival of hamsters immunised with LigANI vaccines after challenge. Percentage survival conferred by rLigANI-Alhydrogel and bacterin against lethal challenge was significant (p < 0.05) in comparison to negative control group. Survival curves were compared using the Mantel-Cox test (LigANI, non-identical carboxy-terminus portion of LigA protein; rLigANI-Alhydrogel, recombinant LigANI in 15% Alhydrogel).

Fig. 1 : cytotoxic effect of carbon nanotubes in CHO-K1 cells. The cytotoxicity of carbon nanotubes was assessed using MTT assay. The inhibition rate was expressed as the optical density ratio of treated cells compared to the negative control cells (only medium). The positive control cells were treated with 1% dimethyl sulfoxide. The data are expressed as mean ± standard deviation of three independent experiments. Asterisks indicate significant differences (p < 0.001) compared to the positive control.

Fig. 2 : induction of IgG antibody response in hamsters immunised with different rLigANI vaccine preparations evaluated by ELISA. Values presented are means ± standard deviation of two independent experiments. Asterisks indicate significant differences (p < 0.05) compared to rLigANI-PBS group [rLigANI- phosphate-buffered saline (PBS), recombinant non-identical carboxy-terminus portion of LigA protein in PBS].

Fig. 3 : isotyping of anti-rLigANI IgG subclasses. The data represent the difference between mean absorbance of hamster sera collected at day 28 and mean absorbance of hamster pre-immune sera at day 0. Results are expressed as mean absorbance ± standard deviation of pooled serum samples assayed in triplicates in two independent experiments. Asterisks indicate significant differences (p < 0.05) compared to rLigANI- phosphate-buffered saline (PBS) group (*) or compared to rLigANI-Alhydrogel group (**) (rLigANI-PBS, recombinant non-identical carboxy-terminus portion of LigA protein in PBS; rLigANI-Alhydrogel, rLigANI in 15% Alhydrogel).

Fig. 4 : survival of hamsters immunised with LigANI vaccines after challenge. Percentage survival conferred by rLigANI-Alhydrogel and bacterin against lethal challenge was significant (p < 0.05) in comparison to negative control group. Survival curves were compared using the Mantel-Cox test (LigANI, non-identical carboxy-terminus portion of LigA protein; rLigANI-Alhydrogel, recombinant LigANI in 15% Alhydrogel).

How to cite

copy

Instituto Oswaldo Cruz, Ministério da Saúde Av. Brasil, 4365 - Pavilhão Mourisco, Manguinhos, 21040-900 Rio de Janeiro RJ Brazil, Tel.: (55 21) 2562-1222, Fax: (55 21) 2562 1220 - Rio de Janeiro - RJ - Brazil
E-mail: memorias@fiocruz.br

Acompanhe os números deste periódico no seu leitor de RSS

Versão para download de PDF

PDF

Inglês

Versões e tradução automática

Versão original do texto

English

Tradução automática

Google Translator
Microsoft Translator

Como citar

RIS

BIBTEX

Outros formatos de citação e exportação:

SciELO - Scientific Electronic Library Online
Rua Dr. Diogo de Faria, 1087 – 9º andar – Vila Clementino 04037-003 São Paulo/SP - Brasil
E-mail: scielo@scielo.org

Leia a Declaração de Acesso Aberto

Métricas

Immune response in hamsters immunised with a recombinant fragment of LigA from <i>Leptospira interrogans</i>, associated with carrier molecules

PlumX

Mensagem

[1] + Corresponding author: daianehartwig@gmail.com