Journal: Genes to cells : devoted to molecular & cellular mechanisms

Article Title: RNA plasticity and selectivity applicable to therapeutics and novel biosensor development.

doi: 10.1111/j.1365-2443.2012.01596.x

Figure Lengend Snippet: Figure 3 Neutralizing anti-interleukin (IL)-17A aptamer (Ishiguro et al. 2011). (A) Secondary structure of Apt21-2, predicted by M-fold. Circles denote 2¢-fluoro-modified pyrimidines. (B) Suppression of IL-17A-induced signaling pathways in normal human dermal fibroblasts (NHDF cells) by Apt21-2. NHDF cells were treated with human (h)IL-17A (40 ng ⁄ mL) with random RNA pool (control) or Apt21-2 RNA (30 nM) and analyzed by Western blotting using the indicated antibodies to detect phosphoryla- tion levels. (C) IL-6 expression affected by Apt21-2 in NHDF cells. hIL-17A was preincubated with Apt21-2 or an anti-hIL-17A antibody at different concentrations and added to NHDF cell culture. After 24-h incubation, the amount of IL-6 secreted to the medium was assessed by ELISA.

Article Snippet: Of note is that under 1.3-nM hIL-17A conditions, Apt21-2 exhibited an IC50 range of 2–3 nM, whereas an available neutralizing anti-hIL-17A monoclonal antibody (mAb317; R&D Systems) had an IC50 of 0 20 40 60 80 100 120 1 10 100 1000 IL -6 ( p g /m L ) Aptamer and antibody concentration (nM) Aptamer Antibody Control (A) (B) (C) Figure 3 Neutralizing anti-interleukin (IL)-17A aptamer (Ishiguro et al. 2011). (A) Secondary structure of Apt21-2, predicted by M-fold.

Techniques: Protein-Protein interactions, Control, Western Blot, Expressing, Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Genes to cells : devoted to molecular & cellular mechanisms

Article Title: RNA plasticity and selectivity applicable to therapeutics and novel biosensor development.

doi: 10.1111/j.1365-2443.2012.01596.x

Figure Lengend Snippet: Figure 4 Attenuation of autoimmunity in mouse models by anti-IL-17A aptamer (Ishiguro et al. 2011). (A) Suppression of experimental autoimmune encephalitis (EAE) development by Apt21-2 (Ishiguro et al. 2011). Wild-type mice (n = 10 each) were immunized with myelin oligodendrocyte protein (MOG35–55) peptide in complete Freund’s adjuvant, and PEG21-2idT (0, 1, 3 and 10 mg ⁄ kg) was administered i.p. every other day after immunization. EAE clinical scores for vehicle and PEG21-2idT–administered mice. Values are the mean and SEM of 10 mice per group. (B) PEG21-2idT treat- ment suppresses development of glucose-6-phosphate isomer- ase (GPI)-induced arthritis. DBA ⁄ 1 mice were immunized with 300 lg of mouse GPI, and the development of arthritis was monitored visually and scored on a scale of 0–2. Values are the mean and SEM of 10 mice per group.

Article Snippet: Of note is that under 1.3-nM hIL-17A conditions, Apt21-2 exhibited an IC50 range of 2–3 nM, whereas an available neutralizing anti-hIL-17A monoclonal antibody (mAb317; R&D Systems) had an IC50 of 0 20 40 60 80 100 120 1 10 100 1000 IL -6 ( p g /m L ) Aptamer and antibody concentration (nM) Aptamer Antibody Control (A) (B) (C) Figure 3 Neutralizing anti-interleukin (IL)-17A aptamer (Ishiguro et al. 2011). (A) Secondary structure of Apt21-2, predicted by M-fold.

Techniques: Adjuvant

Journal: PLOS Pathogens

Article Title: High-density binding to Plasmodium falciparum circumsporozoite protein repeats by inhibitory antibody elicited in mouse with human immunoglobulin repertoire

doi: 10.1371/journal.ppat.1010999

Figure Lengend Snippet: ( A ) Affinity of mAb 850 in comparison to mAbs 1210 and 317 as measured by SPR. ( B ) Capacity of mAb 850 to inhibit the hepatocyte traversal activity of Pf sporozoites in vitro , as compared to mAb 317 . mAbs 850 and 317 were tested at 0.5 μg/mL (left panel) and 1.0 μg/mL (right panel), while humanized versions of murine mAbs 2A10 and mGO53 were used at 100 μg/mL as technical positive and negative controls, respectively. ( C ) mAb 850 displayed the highest affinity to NANP repeats and highest inhibition of Pf sporozoites at 0.5 μg/mL among isolated Kymab antibodies and mAb 317. Affinity and inhibition values are displayed as geometric and arithmetic means, respectively, while error bars represent standard error of the mean (SEM) ( see also ). ( D ) Assessment of antibody in vivo protection. Parasite liver burden load was measured by bioluminescence of Pb sporozoites expressing luciferase-conjugated PfCSP after passive transfer of 100 μg of antibody in mice (N = 5). Numbers above the data points indicate the percent inhibition of the mean parasite burden relative to that of naïve infected (NI) control mice (i.e., % inhibition). Inhibition of mAb 850 was compared to mAb 317 and negative control mAb 1245. NNI—naïve non-infected control mice. Red dashed line in A denotes limit of detection. Black horizontal lines in B – C indicate geometric means. P values were calculated by Bonferroni multiple-comparisons test ( B and C ) and Mann–Whitney test ( A ). *P < 0.05; ****P < 0.0001; n.s., statistically non-significant differences.

Article Snippet: Interestingly, mAb 850 also appeared to be more cross-reactive than mAbs 1210 and 317, with higher affinity to N-terminal peptides KQPADGNPDPNANPN, NPDPNANPNVDPNANP and NVDPNANPNVDPNANPNVDP ( and Figs ). mAb 850 displayed the highest inhibition of Pf traversal of hepatocytes in vitro at 0.5 μg/mL (40.2%), compared to other Kymab IGHV3-33/IGKV1-5- encoded antibodies and potent mAb 317 (28.4%) ( Figs and and S ).

Techniques: Activity Assay, In Vitro, Inhibition, Isolation, In Vivo, Expressing, Luciferase, Infection, Negative Control, MANN-WHITNEY