Intramuscular Matrix-M-adjuvanted virosomal H5N1 vaccine induces high frequencies of multifunctional Th1 CD4+ cells and strong antibody responses in mice

Madhun, Abdullah Sami; Haaheim, Lars R.; Nilsen, Mona V.; Cox, Rebecca Jane

doi:10.1016/j.vaccine.2009.09.044

Cited by 57 publications

(66 citation statements)

References 54 publications

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“…As very few memory T cells will be sustained from a single IFN‐γ producer, a vaccine that induces mainly this response will probably not elicit protective immunity 51 . Interestingly, intranasal immunisation with the pandemic H5N1 virosomal vaccine adjuvanted with matrix M induced much lower frequencies of TNF‐α + /IL‐2 + CD4 + T cells in the mouse spleen 30 compared with the current study. This may be attributed to the differences in the vaccine formulations and the adjuvants used in the two studies and suggests that chitosan may be a better inducer of a multifunctional CD4 + T‐cell response compared with matrix M following intranasal immunisation.…”

Section: Discussioncontrasting

confidence: 65%

“…The influenza‐specific serum IgG, IgG1, IgG2a, IgA and IgM antibodies were quantified using the ELISA, as previously described 30 . Briefly, ELISA plates were coated with inactivated WV influenza H5N1 (RG‐14) antigen or, for the standard curve, capture antibody goat anti‐mouse IgA (catalogue number 1040‐01), IgG (1030‐01), IgG1 (1070‐01) or IgG2a (1080‐01) (Southern Biotechnology, Birmingham, AL, USA).…”

Section: Methodsmentioning

confidence: 99%

“…Cells were then stained for CD3, CD4, CD8 and intracellular cytokine production (IFN‐γ, IL‐2, TNF‐α), and live lymphocytes were acquired using a BD FACSCanto flow cytometer. Data were analysed using FlowJo v8·8·6 software (Tree Star, Ashland, OR, USA), Pestle and spice 4.0 (Mario Roederer, Vaccine Research Centre, NIH, Bethesda, MD, USA), and multifunctional T cells were identified 30 . T cells were classified based on cytokine secretion as single producers (one cytokine), double producers (two cytokines) and multifunctional triple producers (simultaneously secreting IFN‐γ, IL‐2 and TNF‐α).…”

Section: Methodsmentioning

confidence: 99%

“…The Bio‐plex cytokine assay was performed to quantify the concentration of cytokines secreted in the supernatants from stimulated spleen cells as described earlier 30 except that the cytokine secretion was investigated 3 weeks after the second vaccination and that the 7‐plex Group 1 Mouse kit (Bio‐Rad, Cat. #M6000006V7, Bio‐Rad, Gladesville, NSW, Australia) was used to detect cytokines (IL‐2, IFN‐γ, IL‐4, IL‐5, IL‐10 and IL‐17) secreted by spleen cells.…”

Section: Methodsmentioning

confidence: 99%

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The mucosal and systemic immune responses elicited by a chitosan‐adjuvanted intranasal influenza H5N1 vaccine

Svindland

Jul-Larsen

Pathirana

et al. 2011

Influenza Resp Viruses

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Please cite this paper as: Svindland et al. The mucosal and systemic immune responses elicited by a chitosan‐adjuvanted intranasal influenza H5N1 vaccine. Influenza and Other Respiratory Viruses DOI:10.1111/j.1750‐2659.2011.00271.x. Background Development of influenza vaccines that induce mucosal immunity has been highlighted by the World Health Organisation as a priority (Vaccine 2005;23:1529). Dose‐sparing strategies and an efficient mass‐vaccination regime will be paramount to reduce the morbidity and mortality of a future H5N1 pandemic. Objectives This study has investigated the immune response and the dose‐sparing potential of a chitosan‐adjuvanted intranasal H5N1 (RG‐14) subunit (SU) vaccine in a mouse model. Methods Groups of mice were intranasally immunised once or twice with a chitosan (5 mg/ml)‐adjuvanted SU vaccine [7·5, 15 or 30 μg haemagglutinin (HA)] or with a non‐adjuvanted SU vaccine (30 μg HA). For comparison, another group of mice were intranasally immunised with a whole H5N1 (RG‐14) virus (WV) vaccine (15 μg HA), and the control group consisted of unimmunised mice. Results The chitosan‐adjuvanted SU vaccine induced an immune response superior to that of the non‐adjuvanted SU vaccine. Compared with the non‐adjuvanted SU group, the chitosan‐adjuvanted SU vaccine elicited higher numbers of influenza‐specific antibody‐secreting cells (ASCs), higher concentrations of local and systemic antibodies and correspondingly an improved haemagglutination inhibition (HI) and single radial haemolysis (SRH) response against both the homologous vaccine strain and drifted H5 strains. We measured a mixed T‐helper 1/T‐helper 2 cytokine response in the chitosan‐adjuvanted SU groups, and these groups had an increased percentage of virus‐specific CD4+ T cells producing two Thelper 1 (Th1) cytokines simultaneously compared with the non‐adjuvanted SU group. Overall, the WV vaccine induced higher antibody concentrations in sera and an HI and SRH response similar to that of the chitosan‐adjuvanted SU vaccine. Furthermore, the WV vaccine formulation showed a stronger bias towards a T‐helper 1 profile than the SU vaccine and elicited the highest frequencies of CD4+ Th1 cells simultaneously secreting three different cytokines (INFγ+, IL2+ and INFα+). As expected, two immunisations gave a better immune response than one in all groups. The control group had very low or not detectable results in the performed immunoassays. Conclusion The cross‐clade serum reactivity, improved B‐ and T‐cell responses and dose‐sparing potential of chitosan show that a chitosan‐adjuvanted intranasal influenza vaccine is a promising candidate vaccine for further preclinical development.

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Section: Discussioncontrasting

confidence: 65%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

The mucosal and systemic immune responses elicited by a chitosan‐adjuvanted intranasal influenza H5N1 vaccine

Svindland

Jul-Larsen

Pathirana

et al. 2011

Influenza Resp Viruses

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View full text Add to dashboard Cite

show abstract

“…Similarly, immunization of patients with a mixture of HPV16 E6E7 fusion protein and ISCOMATRIX adjuvant induced antigen specific cell mediated immunity in terms of antibody formation, delayed type hypersensitivity, in vitro cytokine release, and CD8 T cell responses [163]. To mitigate the potential safety issues related to ISCOMA-TRIX, Matrix M, the particles made of two selected and purified fractions of saponin, was developed and found to be effective to initiate strong immediate and long-term humoral immune response for influenza H5N1 vaccine with a balanced Th1/Th2 cytokine profile and high crossreactivity against drifted H5N1 viruses in mice [164].…”

Section: Iscomatrixmentioning

confidence: 99%