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Inicio Vacunas (English Edition) Neisseria meningitidis serogroup B lipopolysaccharides induce a lower pro-inflam...
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Vol. 19. Issue 2.
Pages 52-60 (July - December 2018)
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Vol. 19. Issue 2.
Pages 52-60 (July - December 2018)
Original article
Neisseria meningitidis serogroup B lipopolysaccharides induce a lower pro-inflammatory effect within the proteoliposome used in Cuban anti-meningococcal vaccines
Los lipopolisacáridos de Neisseria meningitidis serogrupo B inducen un reducido efecto proinflamatorio dentro del proteoliposoma usado en vacunas cubanas antimeningocócicas
O. Péreza,*, S. Grahamb, M. Lastrea, C.D. Ellisb,c, R.R. Pupoa, D. Tellez-Martínezd, A. Batista-Duharted
a Department of Immunology, University of Medical Science, Havana, Cuba
b Department of Microbiology and Immunology, University of Newcastle upon Tyne, The Medical School, Framlington Place, Newcastle upon Tyne NE2 4HH, UK
c University of Sheffield, D Floor, School of Medicine and Biomedical Sciences, Beech Hill Road, Sheffield S10 2RX, UK
d São Paulo State University (UNESP), School of Pharmaceutical Sciences, Department of Clinical Analysis, Araraquara, SP, Brazil
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Abstract

Neisseria meningitidis outer membrane vesicles or proteoliposomes (PLs) have been used as vaccines and adjuvants. Despite the presence of potentially toxic amounts of lipopolysaccharide (LPS), they have been shown to be safe, well tolerated, and immunogenic. This suggests that LPS-PL may have reduced LPS toxicity. In this study, it is shown that the ability of PL to induce pro-inflammatory cytokine production in human U937 histiocytic cell line is significantly lower than that of an equivalent concentration of purified LPS. This demonstrates that certain components or physical properties of PL reduce the pro-inflammatory activity of their endogenous LPS. To investigate the mechanisms responsible for this protective effect, PLs were fractionated and an assessment was made of the ability of the resulting fractions to induce inflammatory cytokine expression. Several individual PLs fractions were more potent inducers of pro-inflammatory cytokine production than the non-fractionated PLs. The majority of the pro-inflammatory activities appeared to be mediated by the presence of LPS in the fractions, as shown by the ability of an anti-CD14 antibody to block it. However, in two PL fractions, the production of IL-8, and to a lesser extent IL-6, was not inhibited by anti-CD14 treatment, indicating that pro-inflammatory components other than LPS could also be present in PL. Eight proteins present in the fractions were identified by n-terminal sequencing. The results of this study suggest that two of them, PorB, and in particular, the RmpM protein, may also contribute to the pro-inflammatory activity of N. meningitidis PL. These results could support the development of PLs as vaccine adjuvant.

Keywords:
Proteoliposome
Neisseria meningitidis
LPS
Pro-inflammatory cytokines
Adjuvant
Resumen

Las vesículas de la membrana externa de Neisseria meningitidis o proteoliposomas (PL) se utilizan como vacunas y adyuvantes. A pesar de la presencia de cantidades potencialmente tóxicas de lipopolisacárido (LPS) en estas vesículas, se ha demostrado que las vacunas que las utilizan son bien toleradas e inmunogénicas. Esto sugiere que los LPS en PL son menos tóxicos. En el trabajo mostramos que la capacidad de PL para inducir la producción de citoquinas pro-inflamatorias en la línea celular histiocítica U937 humana es significativamente menor que la de una concentración equivalente de LPS purificado, confirmando así que ciertos componentes o propiedades físicas de PL reducen la actividad pro-inflamatoria de sus LPS endógenos. Para investigar los mecanismos responsables de este efecto protector, los PL fueron fraccionados y se evaluó la capacidad de las fracciones resultantes para inducir la expresión de citoquinas inflamatorias. Varias fracciones de PL individuales indujeron la producción de citoquinas pro-inflamatorias en mayor proporción que los PL no fraccionados. La mayoría de las actividades pro-inflamatorias parecían estar mediadas por la presencia de LPS en las fracciones, como se muestra por la capacidad de un anticuerpo anti-CD14 para bloquearlo. Sin embargo, en 2 fracciones de PL, la producción de IL-8 y en menor grado IL-6 no fueron inhibidos por el tratamiento anti-CD14, lo que indica que también podrían estar presentes en los PL componentes proinflamatorios distintos de los LPS. Se identificaron 8 proteínas presentes en las fracciones mediante secuenciación n-terminal. Nuestros resultados sugieren que 2 de ellos PorB y en particular la proteína RmpM también pueden contribuir a la actividad pro-inflamatoria del PL de N. meningitidis. Nuestros resultados apoyan el desarrollo de los PL como adyuvantes en vacunas.

Palabras clave:
Proteoliposoma
Neisseria meningitidis
LPS
Citocinas proinflamatorias
Adyuvante

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