Journal: Microbial biotechnology

Article Title: Rectal single dose immunization of mice with Escherichia coli O157:H7 bacterial ghosts induces efficient humoral and cellular immune responses and protects against the lethal heterologous challenge

doi: 10.1111/j.1751-7915.2011.00316.x

Figure Lengend Snippet: Cellular immune responses stimulated in mice after the immunizations with EHEC O157:H7 (N°CIP 105282) BGs. Mice were immunized as described in Fig. 1 . Spleen cells (1 × 10 5 per well) were incubated for 16 h in the presence of EHEC O157:H7 BGs (specific Ag); measles inactivated Ag (strain Edmonston, non‐specific Ag) or as a negative control spleen cells were left without stimulation. The numbers of IFN‐γ‐producing cells were determined by ELISPOT assays. Rectal immunization with EHEC O157:H7 BGs effectively increased the number of IFN‐γ‐producing spleen cells after recognition of specific Ag (A). The stimulatory effect of the boost immunization was detected on day 42 (14 days after the second immunization) and remained significant until the end of experiment (B). Data represent the mean of three independent experiments ± SD (three mice per time point). P ‐values < 0.05 were considered significant and are indicated with asterisks (* P < 0.05; ** P < 0.01; *** P < 0.001). a, day of the first immunization in all groups; b, day of the second immunization in groups B1 and D1; c, day of the challenge.

Article Snippet: The number of IFN‐γ‐producing cells was determined by Mouse IFN‐γ ELISpot Kit (R&D Systems, Inc, Minneapolis, USA) following the manufacturer's instructions.

Techniques: Incubation, Negative Control, Enzyme-linked Immunospot

Journal: Gene Therapy

Article Title: Significant alterations of biodistribution and immune responses in Balb/c mice administered with adenovirus targeted to CD40(+) cells

doi: 10.1038/sj.gt.3303085

Figure Lengend Snippet: Determination of interferon-γ (IFN-γ) and interleukin-2 (IL-2) in mice receiving either Ad-NP or tAd-NP. The mice were administrated with either Ad-NP (open bar) or tAd-N (filled bar) and boosted 4 weeks later. The splenocytes were then harvested on days 0, 15 and 30 for the analyses of Ag-specific IFN-γ or IL-2 using ELISPOT as described in the ‘Materials and methods’ section. Data represent spots per 10 5 splenocytes stimulated by 10 μg ml −1 of NP. ( a ) IFN-γ levels in mice injected with either Ad-NP or tAd-NP (i.d. route); ( b ) IL-2 levels in mice injected with either Ad-NP or tAd-NP (i.d. route); ( c ) IFN-γ levels in mice injected with either Ad-NP or tAd-NP through the i.n. route); ( d ) IL-2 levels in mice injected with either Ad-NP or tAd-NP (i.n. route). All data represent the means±s.d. from four mice. Asterisks ( * ) indicate significant difference between Ad-NP and tAd-NP (same below). Statistical comparisons were conducted with the use of a two-tailed t -test, with P <0.05 being considered significant. Open bar depicts Ad-NP, while filled bar designates tAd-NP.

Article Snippet: Anti-CD40 mAb, rh-IL-4, recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF), the ELISPOT kits with various controls, including IFN-γ (EL485) and IL-2 (EL402), were purchased from R&D systems, Minneapolis, MN, USA.

Techniques: Enzyme-linked Immunospot, Injection, Two Tailed Test