Expression of iron regulatory genes in PBMCs isolated from unvaccinated controllers and progressors at post-mortem.

<p>Gene expression of a) ferritin heavy chain b) solute carrier family 11 member A1, and c) solute carrier family 11 member A2, d) transferrin receptor 1 and e) heme oxygenase 1 in unstimulated PBMCs isolated at post-mortem. The fold change from naïve baseline levels was determined using RT-PCR. To determine statistically significant differences of relative gene expression, a two-sample t-test was performed where * and ** represent P-values of <0.05 and <0.01, respectively. The data were from 3 animals. The symbol represents the median and the error bars represent the range.</p

Comparison of gene expression profiles generated from microarray and RT-PCR analysis.

<p>RT-PCR was used to validate microarray expression data. The symbol represents the median and the error bars represent the range. Black triangles represent fold change in expression from naïve time-points as determined by microarray analysis; white triangles represent fold change in expression from naïve time-points as determined by RT-PCR analysis. * = differentially expressed (p<0.05).</p

Differentially expressed genes in PPD-stimulated PBMCs isolated from cynomolgus vaccinated controllers 8 weeks post-BCG vaccination.

<p>Fold change indicates the change in gene expression in PPD-stimulated PBMCs isolated at 8 weeks post-BCG vaccination compared with PPD-stimulated PBMCs isolated pre-vaccination when the animals were naïve (n = 6).</p

Histological findings in spleen, liver, testis and heart of A129 mice infected with ZIKV.

<p>Histological findings in spleen, liver, testis and heart of A129 mice infected with ZIKV.</p

Histological findings in brains of A129 mice infected with ZIKV.

<p>Histological findings in brains of A129 mice infected with ZIKV.</p

Clinical data from A129 mice challenged with different doses of ZIKV^AF.

<p>6–8 week old A129 mice were subcutaneously challenged with 10⁶, 10⁵, 10⁴, 10³, 10² or 10 pfu ZIKV^AF virus. (A) Kaplin-Meier survival plot. (B) Differences in weight compared to date of challenge. (C) Clinical score, with numerical values given as follows: 0, normal; 2, ruffled fur; 3, lethargy, pinched, hunched, wasp waisted; 5, laboured breathing, rapid breathing, inactive, neurological; and 10, immobile. Graphs B and C show the mean values with error bars denoting standard error. Group sizes were n = 5.</p

Genetic sequence similarities between ZIKV strains using the study.

<p>Genetic sequence similarities between ZIKV strains using the study.</p

Histological and RNA <i>in situ</i> hybridisation findings in the brain of ZIKV-challenge A129 mice.

<p>(A) Scattered nuclear fragmentation in the hippocampus (Animal 86756, 10⁶ pfu ZIKV^AS, day 7). (B) Perivascular cuffing by mononuclear cells (Animal 86762, 10⁶ pfu ZIKV^AS, day 7). (C) Scattered polymorphonuclear cells (PMNs) in the neuropil, including higher magnification of PMNs (Animal 86722, 10⁶ pfu ZIKV^AF, day 7). (D) Diffuse neuronal degeneration in Ammon’s horn of hippocampus (Animal 86724, 10⁶ ZIKV^AF, day 6). (E) Infiltration of inflammatory cells, mainly mononuclear, in the meninges (Animal 86765, 10 pfu ZIKV^AF, day 7). (F) Occasional scattered cells staining positive for viral RNA in the hippocampus (Animal 86780, 10⁶ pfu ZIKV^AF, day 3). (G) Patchy to diffuse positive staining for viral RNA in the hippocampus (Animal 86783, 10⁶ pfu ZIKV^AF, day 5). (H) Strong positive staining for viral RNA (Animal 86740, 10 pfu ZIKV^AF, day 7). (I) Focus of positively staining cells for viral RNA in sub-ependymal area of the fourth ventricle (Animal 86773, 10⁶ ZIKV^AS, day 5). A-E show sections stained with haematoxylin and eosin (H&E) and F-I show RNA <i>in situ</i> hybridisation images.</p

Clinical data from A129 mice challenged with different strains of ZIKV^AS.

<p>5–8 week old A129 mice were subcutaneously challenged with 10⁶ pfu ZIKV^AS virus. (A) Kaplin-Meier survival plot. (B) Differences in weight compared to date of challenge. (C) Temperature change compared to date of challenge. Graphs B and C show the mean values with error bars denoting standard error. Group sizes were n = 5.</p

Histological and RNA <i>in situ</i> hybridisation findings in the spleen, testes and heart of ZIKV-challenge A129 mice.

<p>(A) Spleen. Poorly defined areas comprising large mononuclear cells within the white pulp (Animal 86783, 10⁶ pfu ZIKV^AF, day 5). Inset, normal spleen with well defined, small germinal centres within the white pulp (Animal 86739, unchallenged). (B) Spleen. Prominent, extra-medullary haematopoiesis in the red pulp. Rectangle, numerous PMNs in the red pulp sinuses (Animal 86724, 10⁶ pfu ZIKV^AF, day 7). (C) Testis. Expansion of the interstitial tissue by proteinaceous fluid, macrophages and PMNs. Inset, higher power image of area within white square (Animal 86772, 10 pfu ZIKV^AF, day 5). (D) Testis. Mild infiltration of PMNs into the interstitial space with positive viral staining (Animal 86779, 10⁶ ZIKV^AF, day 3). (E) Testis. Positive staining of virally infected cells focally within the walls of the seminiferous tubules (white arrows) as well as within the interstitium (Animal 86784, 10 pfu ZIKV^AF, day 7). (F) Testis. Epididymis with positive staining of cells in lumen and epithelium of the efferent ductules as well as the interstitium (Animal 86784, 10 pfu ZIKV^AF, day 7). (G) Testis. Positive staining of cells in the necrotic seminiferous tubules (Animal 86744, 10⁶ pfu ZIKV^AS, day 14). (H) Testis. Intra-tubular and interstitial cell staining (Animal 86757, 10⁶ pfu ZIKV^AS, day 7). (I) Heart. Infiltration of myocardium by macrophages and PMNs (Animal 86779, 10⁶ ZIKV^AF, day 3). (J) Heart. Infiltration of an atrio-ventricular valve by macrophages and PMNs (Animal 86780, 10⁶ pfu ZIKV^AF, day 3). A-C and I-J show sections stained with haematoxylin and eosin (H&E) and D-H show RNA <i>in situ</i> hybridisation images.</p