RSV- and BPZE1-induced signaling pathways in MDDC.

<p><b>A</b>: Total cell lysates were prepared from MDDC treated with BPZE1 (100∶1), RSV (MOI of 1) or both at the indicated time-points. Mock-infected cells were added as control. Proteins were resolved by 10% SDS-PAGE and Western blot analysis was performed to detect the phosphorylation of STAT1, IkBα, ERK1/2 and p38. Human phosphorylated β actin was used to normalize the results. Data are from one representative of three independent experiments performed with MDDC obtained from different donors. <b>B</b>: Histograms represent means of the relative optical density of phosphorylated proteins from three independent experiments. Error bars represent SEM.</p

Impact of BPZE1 on RSV-induced T lymphocyte polarization.

<p>MDDC were treated with BPZE1 (bacteria/cell ratio of 100∶1), RSV (MOI of 1) or both. Mock-infected cells were added as control. After 48 h, treated MDDC were cultured with purified allogeneic CD3 T cells. On day12, supernatants were collected, and IFN-γ (Th1), IL-5 (Th2) and IL-17 (Th17) were measured by ELISA. Values are expressed as medians with interquartile range of 13 independent experiments performed with MDDC obtained from different donors and expressed as ng/ml (IFNγ) or pg/ml (IL-5 and IL-17). Statistical significant differences are indicated by * (<i>P</i>≤0.0083).</p

Impact of BPZE1 on phenotypic maturation of RSV-infected MDDC.

<p>MDDC were treated with BPZE1 (bacteria/cell ratio 100∶1), RSV (MOI of 1) or both. After 24 h, cells were analyzed for the indicated surface markers associated with a mature phenotype. Mock-infected cells were used as control. Fluorescence data are reported as median fluorescence intensity (MedFI) for CD80 and CD38, and as percentage of positive cells for CD83. Values are expressed as medians with interquartile range of seven independent experiments performed with MDDC obtained from different donors. Statistical significant differences are indicated by * (<i>P</i>≤0.0083).</p

RSV- and BPZE1-induced IL-6, IFNβ, CCL5, IL-12p40 and IL-12p35 gene expression in MDDC.

<p>MDDC were treated with BPZE1 (100∶1), RSV (MOI of 1) or both. Mock-infected cells were added as control. Total RNA was extracted at the indicated time points. Kinetics of mRNA expression for p40 and p35 subunits of IL-12p70, IL-6, IFNβ and CCL5 was evaluated by real-time quantitative RT-PCR. The mRNA transcripts were normalized with respect to the endogenous reference (human β actin) sample. Data were expressed as fold increase (mean ± SEM of four experiments) with respect to mock-treated cells at 5 h.</p

Impact of BPZE1 on IL-10, IL-12p70 and IL-23 production of RSV-infected MDDC.

<p>MDDC were treated with BPZE1 (bacteria/cell ratio of 100∶1), RSV (MOI of 1) or double infected with BPZE1 and RSV. Mock-infected cells were used as control. After 24 h, cytokines released in the culture media were measured by ELISA. Values are expressed as medians with interquartile range from 13 (IL-10 and IL-23) and from 15 (IL-12p70) independent experiments performed with MDDC obtained from different donors and expressed as ng/ml (IL-10) or pg/ml (IL-12p70 and IL-23). Statistical significant differences are indicated by * (<i>P</i>≤0.0083).</p

Impact of BPZE1 on RSV-induced allogeneic T cell proliferation.

<p>MDDC were infected with RSV (MOI of 1), BPZE1 (bacteria/cell ratio of 100∶1), or both. Mock-infected cells were added as control. After 48 h, MDDC were co-cultured with allogeneic purified T cells at increasing T cells/MDDC ratios (5/1, 10/1, 20/1, 40/1) for 6 d. Results are reported as percentages of BrdU positive cells (mean ± SEM of 4 independent experiments performed with MDDC obtained from different donors).</p

Diabetes mellitus Affects Antibody Response to SARS-CoV-2 Vaccination in Older Residents of Long-Term Care Facilities: Data from the GeroCovid Vax Study

   ABSTRACT OBJECTIVE: Type 2 Diabetes mellitus may affect the humoral immune response following vaccinations, but data concerning COVID-19 vaccines are scarce. We evaluated the impact of diabetes mellitus on antibody response to the SARS-CoV-2 vaccination in older long-term care facilities (LTCF) residents and tested for differences according to anti-diabetic treatment. RESEARCH DESIGN AND METHODS: 555 older LTCF residents participating in the GeroCovid Vax study were included for this analysis. SARS-CoV-2 trimeric S Immunoglobulin G (anti-S-IgG) concentrations using chemiluminescent assays were tested before the first dose and after 2- and 6-months. The impact of diabetes on anti-S-IgG levels was evaluated using linear mixed models, which included the interaction between time and the presence of diabetes. A second model considered also diabetes treatment: no insulin therapy (including dietary only or use of oral anti-diabetic agents) and insulin therapy (alone or in combination with oral anti-diabetic agents). RESULTS: The sample's mean age was 82.1 years, 68.1% were women and 25.2% were diabetic. In linear mixed models, the presence of diabetes mellitus was associated with lower anti-S-IgG levels 2 (β=-0.20, 95%CI:-0.34,-0.06) and 6 months (β=-0.22, 95%CI:-0.37,-0.07) after the first vaccine dose. Compared to those without diabetes, diabetic residents not using insulin had lower IgG levels at 2- and 6-month assessments (β=-0.24, 95%CI:-0.43,-0.05, and β=-0.30, 95%CI:-0.50,-0.10, respectively), while no differences were observed for those under insulin.  CONCLUSION: Older LTCF residents with diabetes tended to have weaker antibody response to COVID-19 vaccination. Insulin treatment might buffer this effect and establish a humoral immunity similar to non-diabetic individuals.</p

DataSheet_1_The BNT162b2 vaccine induces humoral and cellular immune memory to SARS-CoV-2 Wuhan strain and the Omicron variant in children 5 to 11 years of age.pdf

SARS-CoV-2 mRNA vaccines prevent severe COVID-19 by generating immune memory, comprising specific antibodies and memory B and T cells. Although children are at low risk of severe COVID-19, the spreading of highly transmissible variants has led to increasing in COVID-19 cases and hospitalizations also in the youngest, but vaccine coverage remains low. Immunogenicity to mRNA vaccines has not been extensively studied in children 5 to 11 years old. In particular, cellular immunity to the wild-type strain (Wuhan) and the cross-reactive response to the Omicron variant of concern has not been investigated. We assessed the humoral and cellular immune response to the SARS-CoV-2 BNT162b2 vaccine in 27 healthy children. We demonstrated that vaccination induced a potent humoral and cellular immune response in all vaccinees. By using spike-specific memory B cells as a measurable imprint of a previous infection, we found that 50% of the children had signs of a past, undiagnosed infection before vaccination. Children with pre-existent immune memory generated significantly increased levels of specific antibodies, and memory T and B cells, directed against not only the wild type virus but also the omicron variant.</p

Entry of Tat or R5 Env-VLPs in MDDCs and block by anti-integrin antibodies.

<p>(<b>A</b>) Entry of wt Tat into MDDCs from 3–10 different donors (depending on the anti-integrin mAbs used), in the presence of mAbs against the indicated integrins alone or combined, a control isotype mAb, or nil (buffer). (<b>B</b>) Entry of cys₂₂ Tat into MDDCs from 3 different donors and block by the combined anti-integrin mAbs versus an Ig control isotype mAb. The percentages of Tat positive cells +/− standard deviations are shown. (<b>C</b>) Entry of VLP-R5Env (BaL) in MDDCs in the presence of Tat and block by anti-integrin mAbs or an Ig control isotype mAb. The percentages of fluorescent cells are shown. A representative experiment out of 4 performed is shown.</p

Tat/Env complex and ternary Tat/Env/αvβ3 complex by modeling-docking calculations.

<p>(<b>A</b>) ribbon representation of the Tat/Env complex showing that the Env CD4 binding site and the RGD domain of Tat are both exposed. Color code: ΔV1-2 Env: blue; Tat: red; Tat-RGD: yellow. (<b>B</b>) Surface representation of the ternary Tat/Env/αvβ3 complex. Color code: ΔV1-2 Env: green; Tat: purple; αvβ3 integrin: grey. See experimental procedures for details.</p