Boosting with serologically distinct recombinant virus increases protective potency and longevity of IFN-γ response.

<p>A) Prime-boost with delayed challenge strategy. B) Lesion scores in mice primed with PR8-LACK_ins and boosted with X31-LACK_ins (PR8/X31); and wild type PR8/X31 virus controls. C) Parasite burdens in PR8/X31 vaccinated mice and virus controls (n = 5 each time point, except n = 4 for wk 10 controls), *<i>p</i> = 0.049. D) Cytokine production following challenge. Spleens and dNL from vaccinated and control mice were assessed by <i>ex vivo</i> ICS for IFN-γ production. Mean, pooled data ± SEM are plotted (n = 3 for non-challenged mice, n = 5 for wk 2 and 6; *<i>p</i> = 0.02, **<i>p</i> = 0.01). Cells have been gated on CD4⁺ CD44⁺.</p

Boosting with serologically distinct recombinant virus increases protective potency of the effector response.

<p>A) Prime-boost and challenge strategy. B) Lesion scores in mice primed with PR8-LACK_ins and boosted with X31-LACK_ins (PR8/X31); and PBS controls. C) Parasite burdens in PR8/X31 vaccinated mice and PBS controls (n = 3 for wk 2 and 6, n = 5 for wk 11), *<i>p</i> = 0.009. D) Cytokine production following challenge. Spleens and dNL from vaccinated and control mice were assessed by <i>ex vivo</i> ICS for IFN-γ production. Mean, pooled data ± SEM are plotted (n = 3 for non-challenged mice, n = 5 for wk 2 and 6); *<i>p</i> = 0.02, **<i>p</i> = 0.03, ***<i>p = </i>0.005, ****<i>p</i> = 0.001, #<i>p = </i>0.02, ##<i>p</i> = 0.002, ###<i>p</i> = 0.03. Cells have been gated on CD4⁺ CD44⁺.</p

Cytokine profile of LACK_158–175 specific CD4⁺ T cells.

<p>Cells were obtained from spleens of naïve control mice or PR8-LACK_ins and PR8-LACK_rep mice on day 10 following priming. Cells were cultured in the presence of LACK_158–173 peptide for 72 h, and culture supernatants were analysed for cytokine production by capture ELISA. Mean, pooled data ± SD are plotted (n = 3, n refers to number of mice) *<i>p</i> = 0.001, **<i>p</i> = 0.003, ***<i>p</i> = 0.009, ****<i>p</i> = 0.01, ND – below detection level.</p

Protective efficacy of influenza-LACK_158–173 vaccination.

<p>A) Lesion scores in mice vaccinated with LACK_ins and PR 8 vector controls (pooled data from 2 independent experiments). B) Parasite burdens in mice vaccinated with LACK_ins (n = 5), PR 8 vector control (n = 5), *<i>p</i> = 0.02. C) Cytokine production following challenge infection. Spleens and dNL from vaccinated and control mice were assessed by <i>ex vivo</i> ICS IFN-γ (*<i>p</i> = 0.006). Mean, pooled data from 2 independent experiments ± SEM are plotted (n = 6 for wk 2, n = 5 for wk 6, n refers to number of mice). Cells have been gated on CD4⁺ CD44⁺.</p

Cytokine production by LACK_158–173-specific CD4⁺ T cells following influenza-LACK_158–173 priming.

<p>A) <i>Ex vivo</i> IFN-γ production by LACK_158–173-specific CD4⁺ T cells. Cells were obtained from spleens of naïve control mice or PR8-LACK_ins and PR8-LACK_rep mice on day 10 following priming. Cells were cultured in the absence or presence of LACK_158–173 peptide for 12 h and assessed for IFN-γ production by ICS. B) <i>In vitro</i> IFN-γ production by LACK_158–173-specific CD4⁺ T cells. Cells were obtained from spleens of naïve control mice or PR8-LACK_ins and PR8-LACK_rep mice on day 10 following priming. Cells were cultured in the absence or presence of LACK_158–173 peptide for 72 h followed by re-stimulation with PMA and ionomycin, and assessed for IFN-γ production by ICS. Numbers in the upper right quadrant refer to percentage of CD4⁺ T cells producing IFN-γ (raw data). Cells have been gated on CD4⁺ CD44⁺, for statistical analyses no peptide (panel A) and isotype control (panel B) values were subtracted from the raw values. Representative data from one mouse are shown (n = 3, n refers to number of mice, *<i>p</i> = 0.004, **<i>p</i> = 0.0002, ***<i>p</i> = 0.006).</p

Cytokine production profile of <i>Leishmania</i> specific T cells following long-term prime-boost immunisation.

<p>Cells were obtained from spleens and dLN of immunised and control mice on wk 2 following challenge. Cells were cultured <i>in vitro</i> in the presence of SLA for 72 h, and culture supernatants were analysed for cytokine production. IFN-γ (Th1) levels in splenocytes culture supernatants, and Th2-type cytokine production in dLN on wk 2 post-infection measured by Bio-Plex Pro Mouse Cytokine assay. Mean, pooled data ± SD are plotted (n = 5).</p

Optimization of 2‑Anilino 4‑Amino Substituted Quinazolines into Potent Antimalarial Agents with Oral in Vivo Activity

Novel antimalarial therapeutics that target multiple stages of the parasite lifecycle are urgently required to tackle the emerging problem of resistance with current drugs. Here, we describe the optimization of the 2-anilino quinazoline class as antimalarial agents. The class, identified from publicly available antimalarial screening data, was optimized to generate lead compounds that possess potent antimalarial activity against <i>P. falciparum</i> parasites comparable to the known antimalarials, chloroquine and mefloquine. During the optimization process, we defined the functionality necessary for activity and improved <i>in vitro</i> metabolism and solubility. The resultant lead compounds possess potent activity against a multidrug resistant strain of <i>P. falciparum</i> and arrest parasites at the ring phase of the asexual stage and also gametocytogensis. Finally, we show that the lead compounds are orally efficacious in a 4 day murine model of malaria disease burden

Optimization of 2‑Anilino 4‑Amino Substituted Quinazolines into Potent Antimalarial Agents with Oral in Vivo Activity

Novel antimalarial therapeutics that target multiple stages of the parasite lifecycle are urgently required to tackle the emerging problem of resistance with current drugs. Here, we describe the optimization of the 2-anilino quinazoline class as antimalarial agents. The class, identified from publicly available antimalarial screening data, was optimized to generate lead compounds that possess potent antimalarial activity against <i>P. falciparum</i> parasites comparable to the known antimalarials, chloroquine and mefloquine. During the optimization process, we defined the functionality necessary for activity and improved <i>in vitro</i> metabolism and solubility. The resultant lead compounds possess potent activity against a multidrug resistant strain of <i>P. falciparum</i> and arrest parasites at the ring phase of the asexual stage and also gametocytogensis. Finally, we show that the lead compounds are orally efficacious in a 4 day murine model of malaria disease burden