Figure 1.
Effects of porcine small intestinal submucosa (SIS) on chemokine and cytokine protein microenvironment in BALB/c peritoneal exudates.
BALB/c mice were injected i.p. with SIS preparations (SIS-H or SIS-M) without the antigens and peritoneal exudates harvested after 24 hr. Control groups were treated with PBS buffer. Peritoneal fluids were assayed to determine chemokine and cytokine expression as detailed in Materials and Methods. Data are expressed as the mean relative intensity relative to positive control of each chemokine or cytokine protein detected using pooled peritoneal fluids of 3 mice per group in duplicate. The result is an average of two separate experiments.
Figure 2.
RT-PCR Microarray analysis of transcriptome profiles of inflammatory genes induced by vaccine adjuvants alone in mouse peritoneum.
Eighty-four genes were assessed and those genes up-regulated (A), or down regulated (B) with an average log2 ratio ≥1.5 were selected and plotted as a Venn diagram. Results for the alum-treated group refer to our previous study [24].
Table 1.
RT-PCR Microarray analysis of transcriptome profiles of inflammatory genes induced by vaccine adjuvants alone SIS-H in mouse peritoneum.
Table 2.
RT-PCR Microarray analysis of transcriptome profiles of inflammatory genes induced by vaccine adjuvants alone SIS-H in mouse peritoneum.
Figure 3.
Effects of SIS adjuvants on anti-ovalbumin antibody response in C57BL/6 mice.
Mouse serum samples were collected on day 5 after (A) 2nd, and (B) 3rd immunizations with OVA plus adjuvants as described under Materials and Methods. Antibody responses were assessed using ELISA, (C) determination of IgG sub-classes of anti-OVA antibodies, induced with OVA containing adjuvants, in sera from the 3rd immunization. Commercial isotyping kits (Southern-Biotech) were used to perform ELISA in serum samples (starting at dilution 1∶1000). The results represent mean ± SD (n = 6 mice per group in two separate experiments). The significance in experimental groups relative to the group given antigen only (no adjuvant group) was calculated at the level of p≤0.05.
Figure 4.
Evaluation of phthalate-KLH antibody response in BALB/c mice.
Mice were immunized with phthalate-KLH conjugate emulsified in different adjuvants. Serum samples were collected as described under Materials and Methods and diluted to 1∶1000 in 0.5% PBS/BSA. Anti-phthalate, anti-KLH, and anti-DNA antibody levels were determined using ELISA as described. The results represent the mean ± SD (n = 6 mice per group in two separate experiments). The significance in experimental groups was determined relative to the group given antigen only (no adjuvant group) at the level of p≤0.05.
Figure 5.
Determination of IgG sub-classes of anti-phthalate antibodies induced with phthalate-KLH conjugates in different adjuvants.
This was done using commercial ELISA isotyping kits with serum samples (at dilution 1∶1000) collected after (A) intraperitoneal, or (B) subcutaneous immunization as described in Materials and Methods. Results represent mean ± SD (n = 6 mice per group in two separate experiments). The significance in experimental groups was determined relative to the group given antigen only (no adjuvant group) at the level of p≤0.05.