Increased activated memory B-cells in the peripheral blood of patients with erythema nodosum leprosum reactions - Fig 1

<p>Gating strategy for B-cell sub populations; (1A) gating B-lymphocytes by FSCA versus CD19, (1B) gating mature B-cells (CD19⁺CD10^-), (1C) memory B-cells obtained by gating mature B-cells for CD27 versus CD21.</p

The median percentage of total B-lymphocytes (CD19⁺), mature B-cells (CD19⁺CD10^-) and naïve B-lymphocytes (CD19⁺CD10^-CD27^-CD21⁺) in the PBMCs of) patients with ENL and LL controls before, during and after treatment respectively (2A,2B,2C); within ENL patients before, during and after treatment respectively (2D,2E,2F).

<p>Statistical test: Mann-Whitney unpaired test (U) (2A, 2B, 2C) and Wilcoxon matched-pairs signed rank test (2D, 2E, 2F). * <i>P≤ 0</i>.<i>05; *</i>* <i>P≤ 0</i>.<i>005;</i> *** <i>P≤ 0</i>.<i>001</i>. Box and whiskers show median ± interquartile range.</p

The median percentage of resting memory B-cells (CD19⁺CD10^-CD27⁺CD21⁺), activated memory B-cells (CD19⁺CD10^-CD27⁺CD21^-) and tissue-like memory B-cells (CD19⁺CD10^-CD27^-CD21^-) in the PBMCs of patients with ENL and LL controls before, during and after treatment respectively (3A,3B,3C); within ENL patients before, during and after treatment respectively (3D,3E,3F).

<p>Statistical test: Mann-Whitney unpaired test (U) (3A,3B, 3C)) and Wilcoxon matched-pairs signed rank test (3D, 3E, 3F)B). * <i>P≤ 0</i>.<i>05; *</i>* <i>P≤ 0</i>.<i>005;</i> *** <i>P≤ 0</i>.<i>001</i>. Box and whiskers show median ± interquartile range.</p

DataSheet_1_Protection from COVID-19 disease in hamsters vaccinated with subunit SARS-CoV-2 S1 mucosal vaccines adjuvanted with different adjuvants.pdf

IntroductionAdjuvant plays an important role in directing the immune responses induced by vaccines. In previous studies, we have shown that a mucosal SARS-CoV-2 S1 subunit vaccine adjuvanted with a combination of CpG, Poly I:C and IL-15 (named CP15) induced effective mucosal and systemic immunity and conferred nearly sterile protection against SARS-CoV-2 viral replication in macaque models.MethodsIn this study, we used a hamster model, which mimics the human scenario and reliably exhibits severe SARS-CoV-2 disease similar to hospitalized patients, to investigate the protection efficacy of the vaccines against COVID-19 disease. We compared the weight loss, viral loads (VLs), and clinical observation scores of three different vaccine regimens. All three regimens consisted of priming/boosting with S1 subunit vaccines, but adjuvanted with alum and/or CP15 administrated by either intramuscular (IM) or intranasal (IN) routes: Group 1 was adjuvanted with alum/alum administrated IM/IM; Group 2 was alum-IM/CP15-IN; and Group 3 was CP15-IM/CP15-IN.ResultsAfter challenge with SARS-CoV-2 WA strain, we found that the alum/CP15 group showed best protection against weight loss, while the CP15 group demonstrated best reduction of oral SARS-CoV-2 VLs, suggesting that the protection profiles were different. Sex differences for VL and clinical scores were observed. Humoral immunity was induced but not correlated with protection. Moreover, S1-specific binding antibody titers against beta, omicron BA.1, and BA.2 variants showed 2.6-, 4.9- and 2.8- fold reduction, respectively, compared to the Wuhan strain.DiscussionOverall, the data suggested that adjuvants in subunit vaccines determine the protection profiles after SARS-CoV-2 infection and that nasal/oral mucosal immunization can protect against systemic COVID-19 disease.</p

Comparison of IFN-γ response to Rv2031 among groups at baseline, 6 and 12 months.

<p>IFN-γ response at baseline (a), 6 months (b) and 12 months (c) following whole blood stimulation with Rv2031 for 48 hours. Filled circles represent patients, filled diamonds represent contacts and filled triangles represent controls. Results are individual responses and cytokine levels are expressed as log₁₀ pg/ml. Horizontal bars are medians. Kruskal-Wallis test with Dunn’s multiple comparisons (a) and Mann-Whitney test (b and c) were used to compare groups. ****p<0.0001.</p

Changes in IL-10 responses to Rv2301 over 12 months in patients and contacts.

<p>IL-10 response over 12 months in patients (a) and contacts (b) following whole blood stimulation with Rv2031 for 48 hours. Filled circles represent baseline levels, filled diamonds represent 6 month levels and filled triangles represent 12 month levels. Results are individual responses and cytokine levels are expressed as log₁₀ pg/ml. Horizontal bars are medians. Freidman test with Dunn’s multiple comparisons was used to compare IL-10 response over time. ****p<0.0001.</p

Comparison of IL-10 response to Rv2031 among groups at baseline, 6 and 12 months.

<p>IL-10 response at baseline (a), 6 months (b) and 12 months (c) following whole blood stimulation with Rv2031 for 48 hours. Filled circles represent patients, filled diamonds represent contacts and filled triangles represent controls. Results are individual responses and cytokine levels are expressed as log₁₀ pg/ml. Horizontal bars are medians. Kruskal-Wallis test with Dunn’s multiple comparisons (a) and Mann-Whitney test (b and c) were used to compare groups. ***p<0.001, ****p<0.0001.</p

Correlation between E6C10 and Rv2031 specific IFN-γ, TNF-α and IL-10.

<p>^Vs. versus</p><p>**p<0.01</p><p>For each group, the correlation between E6C10 and Rv2031 specific IFN-γ, TNF-α and IL-10 responses were investigated using Spearman’s correlation and at baseline, Rv2031-specific cytokine responses were positively and significantly correlated with the corresponding cytokine responses to E6C10.</p><p>Correlation between E6C10 and Rv2031 specific IFN-γ, TNF-α and IL-10.</p

Comparison of TNF-α response to Rv2031 among groups at baseline, 6 and 12 months.

<p>TNF-α response at baseline (a), 6 months (b) and 12 months (c) following whole blood stimulation with Rv2031 for 48 hours. Filled circles represent patients, filled diamonds represent contacts and filled triangles represent controls. Results are individual responses and cytokine levels are expressed as log₁₀ pg/ml. Horizontal bars are medians. Kruskal-Wallis test with Dunn’s multiple comparisons (a) and Mann-Whitney test (b and c) were used to compare groups. ***p<0.001, ****p<0.0001.</p

Changes in IFN-γ response to Rv2031 over 12 months in patients and contacts.

<p>IFN-γ response over 12 months in patients (a) and contacts (b) following whole blood stimulation with Rv2031 for 48 hours. Filled circles represent baseline levels, filled diamonds represent 6 month levels and filled triangles represent 12 month levels. Results are individual responses and cytokine levels are expressed as log₁₀ pg/ml. Horizontal bars are medians. Freidman test with Dunn’s multiple comparisons was used to compare IFN-γ response over time. ****p<0.0001.</p