Inborn errors of immunity associated with atopy (IEIs-A) are a group of inherited monogenic disor... more Inborn errors of immunity associated with atopy (IEIs-A) are a group of inherited monogenic disorders that occur with immune dysregulation and frequent skin involvement. Several pathways are involved in the pathogenesis of these conditions, including immune system defects, alterations of skin barrier and metabolism perturbations. Current technological improvements and the higher accessibility to genetic testing, recently allowed the identification of novel molecular pathways involved in IEIs-A, also informing on potential tailored therapeutic strategies. Compared to other systemic therapy for skin diseases, biologics have the less toxic and the best tolerated profile in the setting of immune dysregulation. Here, we review IEIs-A with skin involvement focusing on the tailored therapeutic approach according to their pathogenetic mechanism.
Pediatric Infectious Disease Journal, Oct 31, 2022
Background: Acute pericarditis/myocarditis is a rare complication of the mRNA-based vaccines and ... more Background: Acute pericarditis/myocarditis is a rare complication of the mRNA-based vaccines and although mostly self-limiting, long-term sequelae remain unclear. Methods: We enrolled all patients admitted to the emergency department between September 2021 and February 2022 meeting the CDC work case definition, with symptoms onset after mRNA-based COVID-19 vaccine. Alternative virologic causes were excluded. Clinical data, laboratory values, cardiologic evaluation, electrocardiogram (ECG), and echocardiogram (ECHO) were collected on admission, at discharge, and during follow-up in all patients. Cardiac Magnetic Resonance (CMR) was performed only in those with signs consistent with myocarditis. Results: We observed 13 patients (11M and 2F), median age 15 years, affected by acute pericarditis/myocarditis after COVID-19 mRNA vaccination (11 after Comirnaty® and 2 after Spikevax®). Symptoms’onset occurred at a median of 5 days (range, 1 to 41 days) after receiving mRNA vaccine (13 Prizer 2 Moderna): 4 patients (31%) after the 1st dose, 6 (46%) after the 2nd, and 3 (23%) after 3rd dose. Increased levels of high-sensitive troponin T (hsTnT) (median 519,5 ng/mL) and N-terminal-pro hormone BNP (NT-proBNP) (median 268 pg/mL) and pathognomonic ECG and ECHO abnormalities were detected. On admission, 7 of 13 (54%) presented with myopericarditis, 3 (23%) with myocarditis, and 3 (23%) with pericarditis; CMR was performed in 5 patients upon pediatric cardiologist prescription and findings were consistent with myocarditis. At 12 weeks of follow-up, all but one patient (92%), still presenting mild pericardial effusion at ECHO, were asymptomatic with normal hsTnT and NT-proBNP levels and ECG. On CMR 6 of 9 patients showed persistent, although decreased, myocardial injury. Higher hsTnT levels on admission significantly correlated with persistent CMR lesions. Conclusion: Evidence of persistent CMR lesions highlights the need for a close and standardized follow-up for those patients who present high hsTnT levels on admission.
Children were initially considered unsusceptible to severe COVID-19. Our knowledge after two year... more Children were initially considered unsusceptible to severe COVID-19. Our knowledge after two years has changed dramatically, but there are still many unknowns. Here, we report the current knowledge about why children generally experience a milder COVID-19 course and highlight research questions about pediatric infection that require answers.
Kawasaki disease (KD) is an acute vasculitis occurring in children <5yo that can result in... more Kawasaki disease (KD) is an acute vasculitis occurring in children <5yo that can result in coronary artery lesions. Our study aims to define novel pathogenetic and diagnostic signatures through flow cytometry and gene expression of T cell subsets. Peripheral Blood Mononuclear Cells of twenty-four KD patients at diagnosis (KD), twelve febrile patients (FC) and eighteen age-matched healthy controls (HC) were collected. Frequency of T regulatory cells (Treg), peripheral T follicular helper cells (pTfh) and maturational T cell subsets (Naïve, Central Memory, CM; Effector Memory, EM; terminally differentiated memory, TEMRA) were assessed by FACS (CytoFLEX-Beckman Coulter). Transcriptional levels of 96-genes in sort-purified CM, Treg and EM have been analysed by Fluidgm (Biomark). We found that KD have lower frequency of CM and EM compared to HC (p=0.003 and p=0.005), lower frequency of pTfh cells compared to HC (p=0,008), and in line with previous literature, lower Treg compared to HC (p=0.07). Instead, FC have a higher frequency of EM (p= 0.002; p=0.001) and TEMRA (p=0.004; p=0.006) and lower Naive (p=0.005; p=0.003) compared with both HC and KD. At transcriptional level, a distinct signature was found in EM of FC, with 26 and 16 differentially expressed genes (DEGs) compared to KD and HC. These preliminary data suggest that KD present a distinct phenotypic and transcriptional T cell signature, suggesting additional diagnostic and pathogenic insights able to distinguish KD from FC. Future studies with a similar multi OMIC approach could help identifying prognostic signature in KD.
Most of the current assays directed at the investigation of HIV reactivation are based on culture... more Most of the current assays directed at the investigation of HIV reactivation are based on cultures of infected cells such as Peripheral Blood Mononuclear Cells (PBMCs) or isolated CD4+ T cells, stimulated in vitro with different activator molecules. The culture media in these in vitro tests lack many age- and donor-specific immunomodulatory components normally found within the autologous plasma. This triggered our interest in understanding the impact that different matrices and cell types have on T cell transcriptional profiles following in vitro culture and stimulation. Methods: Unstimulated or stimulated CD4+ T cells of three young adults with perinatal HIV-infection were isolated from PBMCs before or after culture in RPMI medium or autologous plasma. Transcriptomes were sequenced using Oxford Nanopore technologies. Results: Transcriptional profiles revealed the activation of similar pathways upon stimulation in both media with a higher magnitude of TCR cascade activation in CD4+ ...
Multisystem inflammatory syndrome in children (MIS-C) evolves in some pediatric patients followin... more Multisystem inflammatory syndrome in children (MIS-C) evolves in some pediatric patients following acute infection with SARS-CoV-2 by hitherto unknown mechanisms. Whereas acute-COVID-19 severity and outcome were previously correlated with Notch4 expression on regulatory T (Treg) cells, here we show that the Treg cells in MIS-C are destabilized in association with increased Notch1 expression. Genetic analysis revealed that MIS-C patients were enriched in rare deleterious variant impacting inflammation and autoimmunity pathways, including dominant negative mutations in the Notch1 regulators NUMB and NUMBL. Notch1 signaling in Treg cells induced CD22, leading to their destabilization in an mTORC1 dependent manner and to the promotion of systemic inflammation. These results establish a Notch1-CD22 signaling axis that disrupts Treg cell function in MIS-C and point to distinct immune checkpoints controlled by individual Treg cell Notch receptors that shape the inflammatory outcome in SARS...
Coronavirus disease 2019 in children presents with distinct phenotype in comparison to adults. Ov... more Coronavirus disease 2019 in children presents with distinct phenotype in comparison to adults. Overall, the pediatric infection with a generally milder clinical course of the acute infection compared to adults still faces several unknown aspects. Specifically, the presence of a wide range of inflammatory manifestations, including multisystem inflammatory syndrome in children (MIS-C), myocarditis, and long COVID in the period after infection suggests a particular susceptibility of some children upon severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Albeit peculiar complications such as long covid are less frequent in children compared to adults, research on the relationship between inflammatory syndromes and SARS-CoV-2 is rapidly evolving. Conclusions: new studies and findings continue to emerge, providing further insights into the underlying mechanisms and potential therapeutic strategies. In the present work, we revised current knowledge of the main factor...
Introduction The germinal center immune dynamics leading to protective antibody responses after v... more Introduction The germinal center immune dynamics leading to protective antibody responses after vaccination in humans remains largely unknown. Investigation of such dynamics would provide valuable information regarding the cellular and molecular mechanisms governing the generation of these responses and lead to novel vaccine strategies Materials and methods Paraffin embedded tonsil tissue sections were analyzed by multiplexed confocal microscopy assays and Histo-cytometry. The phenotype of tonsil derived cells was analyzed with polychromatic flow cytometry, flu-specific B cell-responses were detected by ELISPOT to H1N1, H3N2, and B strain and plasmas analyzed with a Luminex multiplex assay. Results 36 children (24 vaccinated with 2015/2016 seasonal influenza vaccine and 12 controls) scheduled for elective tonsillectomy were included. Flow cytometry and Histo-Cytometry analysis showed that vaccination induced the frequency of tonsil CD4 Tfh, especially the PD-1hiCD57lo ones. This pro...
Inborn errors of immunity associated with atopy (IEIs-A) are a group of inherited monogenic disor... more Inborn errors of immunity associated with atopy (IEIs-A) are a group of inherited monogenic disorders that occur with immune dysregulation and frequent skin involvement. Several pathways are involved in the pathogenesis of these conditions, including immune system defects, alterations of skin barrier and metabolism perturbations. Current technological improvements and the higher accessibility to genetic testing, recently allowed the identification of novel molecular pathways involved in IEIs-A, also informing on potential tailored therapeutic strategies. Compared to other systemic therapy for skin diseases, biologics have the less toxic and the best tolerated profile in the setting of immune dysregulation. Here, we review IEIs-A with skin involvement focusing on the tailored therapeutic approach according to their pathogenetic mechanism.
Pediatric Infectious Disease Journal, Oct 31, 2022
Background: Acute pericarditis/myocarditis is a rare complication of the mRNA-based vaccines and ... more Background: Acute pericarditis/myocarditis is a rare complication of the mRNA-based vaccines and although mostly self-limiting, long-term sequelae remain unclear. Methods: We enrolled all patients admitted to the emergency department between September 2021 and February 2022 meeting the CDC work case definition, with symptoms onset after mRNA-based COVID-19 vaccine. Alternative virologic causes were excluded. Clinical data, laboratory values, cardiologic evaluation, electrocardiogram (ECG), and echocardiogram (ECHO) were collected on admission, at discharge, and during follow-up in all patients. Cardiac Magnetic Resonance (CMR) was performed only in those with signs consistent with myocarditis. Results: We observed 13 patients (11M and 2F), median age 15 years, affected by acute pericarditis/myocarditis after COVID-19 mRNA vaccination (11 after Comirnaty® and 2 after Spikevax®). Symptoms’onset occurred at a median of 5 days (range, 1 to 41 days) after receiving mRNA vaccine (13 Prizer 2 Moderna): 4 patients (31%) after the 1st dose, 6 (46%) after the 2nd, and 3 (23%) after 3rd dose. Increased levels of high-sensitive troponin T (hsTnT) (median 519,5 ng/mL) and N-terminal-pro hormone BNP (NT-proBNP) (median 268 pg/mL) and pathognomonic ECG and ECHO abnormalities were detected. On admission, 7 of 13 (54%) presented with myopericarditis, 3 (23%) with myocarditis, and 3 (23%) with pericarditis; CMR was performed in 5 patients upon pediatric cardiologist prescription and findings were consistent with myocarditis. At 12 weeks of follow-up, all but one patient (92%), still presenting mild pericardial effusion at ECHO, were asymptomatic with normal hsTnT and NT-proBNP levels and ECG. On CMR 6 of 9 patients showed persistent, although decreased, myocardial injury. Higher hsTnT levels on admission significantly correlated with persistent CMR lesions. Conclusion: Evidence of persistent CMR lesions highlights the need for a close and standardized follow-up for those patients who present high hsTnT levels on admission.
Children were initially considered unsusceptible to severe COVID-19. Our knowledge after two year... more Children were initially considered unsusceptible to severe COVID-19. Our knowledge after two years has changed dramatically, but there are still many unknowns. Here, we report the current knowledge about why children generally experience a milder COVID-19 course and highlight research questions about pediatric infection that require answers.
Kawasaki disease (KD) is an acute vasculitis occurring in children <5yo that can result in... more Kawasaki disease (KD) is an acute vasculitis occurring in children <5yo that can result in coronary artery lesions. Our study aims to define novel pathogenetic and diagnostic signatures through flow cytometry and gene expression of T cell subsets. Peripheral Blood Mononuclear Cells of twenty-four KD patients at diagnosis (KD), twelve febrile patients (FC) and eighteen age-matched healthy controls (HC) were collected. Frequency of T regulatory cells (Treg), peripheral T follicular helper cells (pTfh) and maturational T cell subsets (Naïve, Central Memory, CM; Effector Memory, EM; terminally differentiated memory, TEMRA) were assessed by FACS (CytoFLEX-Beckman Coulter). Transcriptional levels of 96-genes in sort-purified CM, Treg and EM have been analysed by Fluidgm (Biomark). We found that KD have lower frequency of CM and EM compared to HC (p=0.003 and p=0.005), lower frequency of pTfh cells compared to HC (p=0,008), and in line with previous literature, lower Treg compared to HC (p=0.07). Instead, FC have a higher frequency of EM (p= 0.002; p=0.001) and TEMRA (p=0.004; p=0.006) and lower Naive (p=0.005; p=0.003) compared with both HC and KD. At transcriptional level, a distinct signature was found in EM of FC, with 26 and 16 differentially expressed genes (DEGs) compared to KD and HC. These preliminary data suggest that KD present a distinct phenotypic and transcriptional T cell signature, suggesting additional diagnostic and pathogenic insights able to distinguish KD from FC. Future studies with a similar multi OMIC approach could help identifying prognostic signature in KD.
Most of the current assays directed at the investigation of HIV reactivation are based on culture... more Most of the current assays directed at the investigation of HIV reactivation are based on cultures of infected cells such as Peripheral Blood Mononuclear Cells (PBMCs) or isolated CD4+ T cells, stimulated in vitro with different activator molecules. The culture media in these in vitro tests lack many age- and donor-specific immunomodulatory components normally found within the autologous plasma. This triggered our interest in understanding the impact that different matrices and cell types have on T cell transcriptional profiles following in vitro culture and stimulation. Methods: Unstimulated or stimulated CD4+ T cells of three young adults with perinatal HIV-infection were isolated from PBMCs before or after culture in RPMI medium or autologous plasma. Transcriptomes were sequenced using Oxford Nanopore technologies. Results: Transcriptional profiles revealed the activation of similar pathways upon stimulation in both media with a higher magnitude of TCR cascade activation in CD4+ ...
Multisystem inflammatory syndrome in children (MIS-C) evolves in some pediatric patients followin... more Multisystem inflammatory syndrome in children (MIS-C) evolves in some pediatric patients following acute infection with SARS-CoV-2 by hitherto unknown mechanisms. Whereas acute-COVID-19 severity and outcome were previously correlated with Notch4 expression on regulatory T (Treg) cells, here we show that the Treg cells in MIS-C are destabilized in association with increased Notch1 expression. Genetic analysis revealed that MIS-C patients were enriched in rare deleterious variant impacting inflammation and autoimmunity pathways, including dominant negative mutations in the Notch1 regulators NUMB and NUMBL. Notch1 signaling in Treg cells induced CD22, leading to their destabilization in an mTORC1 dependent manner and to the promotion of systemic inflammation. These results establish a Notch1-CD22 signaling axis that disrupts Treg cell function in MIS-C and point to distinct immune checkpoints controlled by individual Treg cell Notch receptors that shape the inflammatory outcome in SARS...
Coronavirus disease 2019 in children presents with distinct phenotype in comparison to adults. Ov... more Coronavirus disease 2019 in children presents with distinct phenotype in comparison to adults. Overall, the pediatric infection with a generally milder clinical course of the acute infection compared to adults still faces several unknown aspects. Specifically, the presence of a wide range of inflammatory manifestations, including multisystem inflammatory syndrome in children (MIS-C), myocarditis, and long COVID in the period after infection suggests a particular susceptibility of some children upon severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Albeit peculiar complications such as long covid are less frequent in children compared to adults, research on the relationship between inflammatory syndromes and SARS-CoV-2 is rapidly evolving. Conclusions: new studies and findings continue to emerge, providing further insights into the underlying mechanisms and potential therapeutic strategies. In the present work, we revised current knowledge of the main factor...
Introduction The germinal center immune dynamics leading to protective antibody responses after v... more Introduction The germinal center immune dynamics leading to protective antibody responses after vaccination in humans remains largely unknown. Investigation of such dynamics would provide valuable information regarding the cellular and molecular mechanisms governing the generation of these responses and lead to novel vaccine strategies Materials and methods Paraffin embedded tonsil tissue sections were analyzed by multiplexed confocal microscopy assays and Histo-cytometry. The phenotype of tonsil derived cells was analyzed with polychromatic flow cytometry, flu-specific B cell-responses were detected by ELISPOT to H1N1, H3N2, and B strain and plasmas analyzed with a Luminex multiplex assay. Results 36 children (24 vaccinated with 2015/2016 seasonal influenza vaccine and 12 controls) scheduled for elective tonsillectomy were included. Flow cytometry and Histo-Cytometry analysis showed that vaccination induced the frequency of tonsil CD4 Tfh, especially the PD-1hiCD57lo ones. This pro...
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