Resting and IFN-γ stimulated monocytes show increased levels of CD100 mRNA compared to macrophages and foam cells (A, B).

<p>Quantitative RT-PCR (qRT-PCR) for CD100 (A) or STAT-1 (B) in PB monocytes, macrophages and foam cells stimulated (+) or not (-) with IFN-γ. Folds relative to non stimulated monocytes (mono-), normalized with GAPDH. (a) * = significant <i>p</i> values (p≤0.05) in the comparisons in unstimulated and stimulated conditions.</p

CD100 is expressed only in endothelia of normal coronary and carotid arteries.

<p>CD100 labeling (E-H) and negative controls (A-D) in normal carotid (A, B, E, F) and coronary (C, D, G, H) artery tissue sections by immunohistochemistry. Positive CD100 staining was observed only in endothelia of normal arteries (arrowheads). Scale bars: 10µm (<i>A</i>, <i>E</i>, <i>C</i> and <i>G</i>) and 50µm (<i>B</i>, <i>F</i>, <i>D</i> and <i>H</i>).</p

Infiltrating cells of the intima in coronary and carotid plaques are CD100 positive.

<p>CD100 labeling (E-H) and negative controls (A-D) in artery tissue sections of carotid plaques (A, B, E, F) and coronary plaques (C, D, G, H) by immunohistochemistry. Positive CD100 staining was observed only in preserved endothelium (arrowhead), and in infiltrating cells in the intima (asterisks). Scale bars: 10µm (<i>A</i>, <i>E</i>, <i>C</i> and <i>G</i>), and 50µm (<i>B</i>, <i>F</i>, <i>D</i> and <i>H</i>).</p

Cultured monocytes express higher amounts of CD100 protein than <i>in vitro</i> differentiated macrophages and foam cells.

<p>CD100 and β-actin protein expression was evaluated in activated T lymphocytes (TL – positive control) and PB monocytes (Mono), macrophages (Mac) and foam cells (Foam) A. Western blot showing CD100 and β-actin bands B. Densitometry of CD100 (sum of 120 and 150kDa bands) and β-actin protein bands, showing CD100/β-actin. Mean ± SD of 3 independent experiments.</p

(A, B): CD36 mRNA is reduced in cultured macrophages treated with oxLDL in the presence of CD100 and IL-10.

<p>Relative quantitative RT-PCR (qRT-PCR) for CD36 in macrophages (<b>A</b>) and foam cells (<b>B</b>) incubated with oxLDL in the presence of CD100 or IL-10. Data normalized with GAPDH and expressed as a ratio (fold) relative to cells incubated only with oxLDL. Mean ± SD of 3 independent experiments (** p≤0.01).</p

<i>In vitro</i> cultured monocytes, macrophages and foam cells express CD100.

<p>Immunofluorescence of PB monocytes (A-D), macrophages (E-H) and foam cells (I-L) showing blue nuclei in DAPI (A, E, I), CD68 (<i>green</i>; B, F, J), CD100 (<i>red</i>; C, G, K) and co-localization of all markers in D, H and L. Scale bars: 5µm.</p

Macrophages and foam cells from atherosclerotic plaques express CD100.

<p>Immunofluorescence of carotid plaques with blue nuclei in DAPI (A), CD68 staining in FITC (<i>green</i>, B), CD100 staining in Alexa 660 (<i>red</i>, C) and co-localization of markers (D). Scale bars: 20µm.</p

image_2_Acute Zika Virus Infection in an Endemic Area Shows Modest Proinflammatory Systemic Immunoactivation and Cytokine-Symptom Associations.jpeg

<p>An early immune response to Zika virus (ZIKV) infection may determine its clinical manifestation and outcome, including neurological effects. However, low-grade and transient viremia limits the prompt diagnosis of acute ZIKV infection. We have investigated the plasma cytokine, chemokine, and growth factor profiles of 36 individuals from an endemic area displaying different symptoms such as exanthema, headache, myalgia, arthralgia, fever, hyperemia, swelling, itching, and nausea during early-phase infection. These profiles were then associated with symptoms, revealing important aspects of the immunopathophysiology of ZIKV infection. The levels of some cytokines/chemokines were significantly higher in acute ZIKV-infected individuals compared to healthy donors, including interferon (IFN) gamma-induced protein 10 (IP-10), regulated on activation, normal T cell expressed and secreted (RANTES), IFN-γ, interleukin (IL)-9, IL-7, IL-5, and IL-1ra, including some with predominantly immunoregulatory activity. Of note, we found that higher levels of IP-10 and IL-5 in ZIKV-infected individuals were strongly associated with exanthema and headache, respectively. Also, higher levels of IL-1ra were associated with subjects with arthralgia, whereas those with fever showed lower levels of granulocyte-colony stimulating factor (G-CSF). No correlation was observed between the number of symptoms and ZIKV viral load. Interestingly, only IP-10 showed significantly decreased levels in the recovery phase. In conclusion, our results indicate that acute ZIKV infection in a larger cohort resident to an endemic area displays a modest systemic immune activation profile, involving both proinflammatory and immunoregulatory cytokines and chemokines that could participate of virus control. In addition, we showed that differential cytokine/chemokine levels are related to specific clinical symptoms, suggesting their participation in underlying mechanisms.</p

Data_Sheet_1_Post-COVID-19 condition: systemic inflammation and low functional exercise capacity.pdf

IntroductionPost-COVID-19 condition (PCC) is characterised by a plethora of symptoms, with fatigue appearing as the most frequently reported. The alterations that drive both the persistent and post-acute disease newly acquired symptoms are not yet fully described. Given the lack of robust knowledge regarding the mechanisms of PCC we have examined the impact of inflammation in PCC, by evaluating serum cytokine profile and its potential involvement in inducing the different symptoms reported.MethodsIn this cross-sectional study, we recruited 227 participants who were hospitalised with acute COVID-19 in 2020 and came back for a follow-up assessment 6–12 months after hospital discharge. The participants were enrolled in two symptomatic groups: Self-Reported Symptoms group (SR, n = 96), who did not present major organ lesions, yet reported several debilitating symptoms such as fatigue, muscle weakness, and persistent loss of sense of smell and taste; and the Self-Reported Symptoms and decreased Pulmonary Function group (SRPF, n = 54), composed by individuals with the same symptoms described by SR, plus diagnosed pulmonary lesions. A Control group (n = 77), with participants with minor complaints following acute COVID-19, was also included in the study. Serum cytokine levels, symptom questionnaires, physical performance tests and general clinical data were obtained in the follow-up assessment.ResultsSRPF presented lower IL-4 concentration compared with Control (q = 0.0018) and with SR (q = 0.030), and lower IFN-α2 serum content compared with Control (q = 0.007). In addition, SRPF presented higher MIP-1β serum concentration compared with SR (q = 0.029). SR presented lower CCL11 (q = 0.012 and q = 0.001, respectively) and MCP-1 levels (q = 0.052 for both) compared with Control and SRPF. SRPF presented lower G-CSF compared to Control (q = 0.014). Female participants in SR showed lower handgrip strength in relation to SRPF (q = 0.0082). Male participants in SR and SRPF needed more time to complete the timed up-and-go test, as compared with men in the Control group (q = 0.0302 and q = 0.0078, respectively). Our results indicate that different PCC symptom profiles are accompanied by distinct inflammatory markers in the circulation. Of particular concern are the lower muscle function findings, with likely long-lasting consequences for health and quality of life, found for both PCC phenotypes.</p