J Infect Dis. received Advax?-adjuvanted vaccine to na?ve mice by immune serum. Enhanced humoral and T-cell responses induced by Advax?-formulated vaccine were sustained 12 months post-immunization. Advax? adjuvant had low reactogenicity and no adverse events were identified. This suggests Advax? adjuvant could be a useful influenza vaccine adjuvant. 0.001) (Fig. 1B) and IgG2a subtypes ( 0.05) (Fig. 1C). This translated into significantly higher hemagglutinin inhibition titers in mice receiving Advax?-adjuvanted TIV ( 0.01) (Fig. 1D) when compared to TIV alone. Open in a separate window Open in a separate window Fig. 1 Co-administration of Advax? adjuvant with influenza vaccine enhances humoral and cellular responses. (ACD) Adult female BALB/c mice (n=5) were immunized intramuscularly twice at a 2-week interval with 40ng HA alone (white bars) or with Advax? 1mg (black bars). Blood samples were collected 2 weeks after the second immunization and IgG (A), IgG1 (B) and IgG2a (C) measured by ELISA. The HI titer was read as the endpoint dilution of serum that completely inhibited hemagglutination and is presented as the log2 titer plus standard error (D). (ECF) BM and spleen were collected from adult BALB/c mice two weeks following the second immunization with PR8 antigen alone (white bars) or with Advax? (black bars). PR8-specific IgG or IgM ASC in BM (E) and spleen (F) were detected by ELISPOT assay using PR8-coated plates. Data show the average ASC frequencies from 11 mice/group. (G) Female BALB/c mice were immunized intramuscularly twice at a 2-week interval with 45ng HA of TIV antigen with or without Advax? adjuvant. Spleens were collected 5 weeks after the second immunization and antigen-specific CD4 and CD8 T-cell proliferation measured by culturing CFSE-labeled splenocytes with Mouse monoclonal to THAP11 TIV antigen for 5 days (n = 6, Varenicline Tartrate mean Varenicline Tartrate + SEM). Asterisks designate significant differences (* 0.05, ** 0.01, *** 0.001). Advax? adjuvant increases antibody secreting B cells To assess whether higher antibody responses correlated with a higher frequency of antibody secreting cells (ASC), influenza-specific antibody secreting cells were measured by ELISPOT in bone marrow and spleen from PR8-immunized mice. Mice immunized with PR8 formulated with Advax? adjuvant had significantly higher frequencies of influenza-specific B cells secreting either IgG or IgM in bone marrow (Fig. 1E) and spleen (Fig. 1F) when compared to mice immunized with PR8 alone. Advax? adjuvant increases T-cell proliferative responses to influenza T-cell help is required for generation of isotype-switched B cells. To assess whether influenza antigen formulated with Advax? adjuvant increased T-cell recall responses, Varenicline Tartrate splenocytes from mice immunized with influenza antigen with or without Advax? adjuvant were labeled with CFSE and then cultured with influenza antigen for 5 days. Mice that had received vaccine formulated with Advax? adjuvant had significantly higher CD4 ( 0.01) and CD8 ( 0.001) T-cell proliferation in response to influenza antigen when compared to mice that received influenza antigen alone (Fig. 1G). Advax?-adjuvanted vaccine induces a mixed Th1 and Th2 cytokine profile Given the increased T-cell proliferation in response to influenza antigen observed in mice immunized with influenza antigen plus Advax? adjuvant, we asked whether Advax? might have imparted a skew towards either a Th1 or Th2 response. Splenocytes from immunized mice were re-stimulated for 3 days with influenza antigen and culture supernatants harvested for cytokine measurement. Splenocytes from mice that received Advax?-adjuvanted vaccine produced significantly higher IL-2, IL-5, IL-6, IFN- and GM-CSF, no change in IL-4 and a non-significant trend towards lower IL-1 and TNF (Fig. 2),.
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