Activated phosphoinositide 3-kinase δ syndrome caused by PIK3CD mutations: expanding the phenotype.

BACKGROUND: Germline heterozygous gain-of-function (GOF) mutations in the PIK3CD gene lead to a rare primary immunodeficiency disease known as activated phosphoinositide 3-kinase (PI3K) δ syndrome type 1(APDS1). Affected patients present a spectrum of clinical manifestations, particularly recurrent respiratory infections and lymphoproliferation, increased levels of serum immunoglobulin (Ig) M, Epstein-Barr virus (EBV) and cytomegalovirus (CMV) viremia. Due to highly heterogeneous phenotypes of APDS1, it is very likely that suspected cases may be misdiagnosed. METHODS: Herein we reported three patients with different clinical presentations but harboring pathogenic variants in PIK3CD gene detected by trio whole-exome sequencing (trio-WES) and confirmed by subsequent Sanger sequencing. RESULTS: Two heterozygous mutations (c.3061G > A, p.E1021K and c.1574 A > G, p.E525G) in PIK3CD (NM_005026.3) were identified by whole exome sequencing (WES) in the three patients. One of two patients with the mutation (c.3061G > A) presented with abdominal pain and diarrhea as the first symptoms, which was due to intussusception caused by multiple polyps of colon. The patient with mutation (c.1574 A > G) had an anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV)-like clinical manifestations, including multisystemic inflammation, acute nephritic syndrome, and positive perinuclear ANCA (p-ANCA), thus the diagnosis of ANCA-AAV was considered. CONCLUSIONS: Our study expands the spectrums of clinical phenotype and genotype of APDS, and demonstrates that WES has a high molecular diagnostic yield for patients with immunodeficiency related symptoms, such as respiratory infections, multiple ecchymosis, ANCA-associated vasculitis, multiple ileocecal polyps, hepatosplenomegaly, and lymphoid hyperplasia. TRIAL REGISTRATION: Retrospectively registered.

In vivo G-CSF treatment activates the GR-SOCS1 axis to suppress IFN-γ secretion by natural killer cells.

Natural killer (NK) cells are lymphocytes that are involved in controlling tumors or microbial infections through the production of interferon gamma (IFN-γ). Granulocyte colony-stimulating factor (G-CSF) inhibits IFN-γ secretion by NK cells, but the mechanism underlying this effect remains unclear. Here, by comparing the multi-omics profiles of human NK cells before and after in vivo G-CSF treatment, we identify a pathway that is activated in response to G-CSF treatment, which suppresses IFN-γ secretion in NK cells. Specifically, glucocorticoid receptors (GRs) activated by G-CSF inhibit secretion of IFN-γ by promoting interactions between SOCS1 promoters and enhancers, as well as increasing the expression of SOCS1. Experiments in mice confirm that G-CSF treatment significantly downregulates IFN-γ secretion and upregulates GR and SOCS1 expression in NK cells. In addition, GR blockade by the antagonist RU486 significantly reverses the effects of G-CSF, demonstrating that GRs upregulate SOCS1 and inhibit the production of IFN-γ by NK cells.

Tidal Breathing Pulmonary Function of Children With Allergic Rhinitis.

In order to investigate the characteristics of tidal breathing pulmonary function in children with allergic rhinitis, and explore its role in the relationship between allergic rhinitis and asthma, we conducted this prospective study from January 4, 2016 to January 30, 2019 in Wuhan children's hospital. In this study, 49 children with simple allergic rhinitis were enrolled in the AR group; 50 children with allergic rhinitis concomitant with asthma were enrolled in the AR&A group; 43 healthy children were recruited in the control group. For individuals in each group, the assessment of tidal breath pulmonary function was performed after enrollment. Then participants in the AR group and control group were followed up for 1 year to observe their frequency of wheezing attacks. The parameters of tI/tE, tPTEF/tE, and VPTEF/VE of AR group were significantly higher than AR&A group (P < 0.001). The reduced proportion of tPTEF/tE and VPTEF/VE. in AR group were higher than that in control group (30.61% vs. 11.63%, P < 0.001; 24.49% vs. 11.63%, P < 0.001, respectively). The proportion of patients with reduced tPTEF/tE and VPTEF/VE who occurred recurrent wheezing was higher than that of patients with normal pulmonary function in AR group(P = 0.008). In conclusion, some children with allergic rhinitis has impaired tidal breathing pulmonary function. Tidal breathing pulmonary function test plays an important role in the diagnosis and assessment of children's airway allergic diseases (AR and asthma).

Human Adenovirus Type 7 Infections in Hubei, China During 2018-2019: Epidemic Features and Genetic Characterization of the Detected Viruses.

Human adenoviruses (HAdVs) type 7 can cause severe respiratory disease. During the period between December 2018 and August 2019, HAdV-7 infection was identified in 129 patients in Wuhan Children's Hospital, Hubei Province, China. Samples were collected from hospitalized children and metagenomic sequencing was applied to detect the HAdV infections. Hemophagocytic lymphohistiocystosis (HLH) related to HAdV infections was observed in some patients clinically and patients were divided into two groups based on this to test the differences among clinical indicators. Genome variation, in silico restriction endonuclease analysis (REA), and phylogenetic analyses were carried out to show the genome characterization of HAdV-7 in this study. It was found that many indicators, such as all blood routine indicators, in patients of the HLH group showed significant levels. In this study, REA revealed that HAdV-7 might belong to genome 7d and genome variation analysis displayed the stable genome of HAdV. HAdV-7 is an ongoing threat to the public, and global surveillance should be established.

Plasmacytoid dendritic cell deficiency in neonates enhances allergic airway inflammation via reduced production of IFN-α.

Allergic asthma, a chronic inflammatory airway disease associated with type 2 cytokines, often originates in early life. Immune responses at an early age exhibit a Th2 cell bias, but the precise mechanisms remain elusive. Plasmacytoid dendritic cells (pDCs), which play a regulatory role in allergic asthma, were shown to be deficient in neonatal mice. We report here that this pDC deficiency renders neonatal mice more susceptible to severe allergic airway inflammation than adult mice in an OVA-induced experimental asthma model. Adoptive transfer of pDCs or administration of IFN-α to neonatal mice prevented the development of allergic inflammation in wild type but not in IFNAR1-/- mice. Similarly, adult mice developed more severe allergic inflammation when pDCs were depleted. The protective effects of pDCs were mediated by the pDC-/IFN-α-mediated negative regulation of the secretion of epithelial cell-derived CCL20, GM-CSF, and IL-33, which in turn impaired the recruitment of cDC2 and ILC2 cells to the airway. In asthmatic patients, the percentage of pDCs and the level of IFN-α were lower in children than in adults. These results indicate that impairment of pDC-epithelial cell crosstalk in neonates is a susceptibility factor for the development of allergen-induced allergic airway inflammation.

[Relationship between serum 25-hydroxyl-vitamin D3 levels and galectin-3 levels in serum and bronchoalveolar lavage fluid in children with asthma].

OBJECTIVE: To investigate the changes of the levels of galectin-3 (Gal-3) in serum and bronchoalveolar lavage fluid (BALF) of children with asthma whose have different serum levels of 25-hydroxyl-vitamin D3[25(OH)D3]. METHODS: Fifty children with asthma between January 2013 and December 2014 were enrolled as the asthma group, and they were classified into 25(OH)D3sufficient (n=7), insufficient (n=12) and deficient subgroups (n=31) according to the serum levels of 25(OH)D3. Twenty children with abnormal airway or tracheal foreign bodies served as the control group. The levels of 25(OH)D3, Gal-3 and total IgE in serum and Gal-3 levels in BALF were measured using ELISA. RESULT: The serum levels of 25(OH)D3in the asthma group were lower than in the control group (P<0.05). The 25(OH)D3deficient subgroup displayed the highest percentages of neutrophils, eosinophils and epithelial cells in BALF, followed by the 25(OH)D3insufficient subgroup and the 25(OH)D3sufficient subgroup (P<0.05). The percentages of neutrophils, eosinophils and epithelial cells in BALF in the three subgroups were all higher than in the control group (P<0.05). In children with asthma, serum levels of 25(OH)D3were negatively correlated with the percentages of neutrophils, eosinophils and epithelial cells in BALF (r=-0.683, -0.795 and -0.670 respectively; P<0.05); and a negative correlation was also seen between serum 25(OH)D3levels and serum Gal-3 and total IgE levels (r=-0.759 and -0.875 respectively; P<0.05). CONCLUSIONS: The children with asthma have low serum levels of 25(OH)D3. 25(OH)D3and Gal-3 may be involved in the airway inflammation and the development of asthma.

Galectin-9 ameliorates respiratory syncytial virus-induced pulmonary immunopathology through regulating the balance between Th17 and regulatory T cells.

Respiratory syncytial virus (RSV) infections are characterized by lung inflammation, mucus hypersecretion, and hyperresponsiveness. CD4+ T cells play a pivotal role in the development of RSV-induced lung pathology. Thus targeting the activation of CD4+ T cell subsets and enhancing regulatory functions of CD4+ T cells could be an effectively therapeutic approach. In the present study, we showed that RSV-induced lung inflammation can be suppressed by lectin family member Galectin-9 (Gal-9), which is identified as a T-cell immunoglobulin- and mucindomain-containing molecule-3 (Tim-3) ligand (L) and the Gal-9/Tim-3 interaction acts as a specific inhibitor of T helper(Th)1 and Th17 immune responses. Tim-3 expression was up-regulated in RSV-infected mice compared to non-infected controls. Therefore, we constructed a recombinant adenoviral (rAAV) 9-Gal-9 adenoviral plasmid, and administered it intranasally into RSV-infected mice for five times at every other day until day 8 post-infection. We found that Gal-9 administration significantly decreased viral load, inhibited mucus production, and diminished severity of lung pathology which were all induced by RSV infection. Complicated mechanisms were involved in these inhibitory effects, including inhibition of Th17 cell production, induction of regulatory cell expansion, as well as alteration of CD8 T-cell apoptosis. Our findings suggest that regulating the function of the Gal-9/Tim-3 pathway will be an effective and safe approach to treat RSV infection in lungs.

[Etiology, clinical features, and diagnosis and treatment of recurrent hemoptysis in children].

OBJECTIVE: To investigate the causes, clinical features, therapy and treatment outcomes of recurrent hemoptysis in children and to improve the skills of pediatricians in the etiological diagnosis and treatment of recurrent hemoptysis in children. METHODS: The clinical, laboratory, and imaging data of 39 children with recurrent hemoptysis between January 1996 and February 2013 were collected to retrospectively analyze the age of onset, etiology, amount of hemoptysis, imaging changes, treatment methods, and follow-up outcomes. RESULTS: In the 39 children, including idiopathic pulmonary hemosiderosis (16 cases, 41%), pulmonary vascular malformation (8 cases, 21%), pulmonary arteriovenous fistula (7 cases, 18%), bronchiectasis (3 cases, 8%), pulmonary tuberculosis (2 cases, 5%), pulmonary cystic fibrosis (2 cases, 5%), and lung tumor (1 case, 3%). The contrast-enhanced lung CT scans and pulmonary and bronchial arteriography revealed varying degrees of lung imaging changes in 35 cases. Of all cases, 51% were classified as degree I, 28% as degree II, and 21% as degree III. All children were treated according to the etiology, with the disease controlled. During 0.5-5 years of follow-up, 3 patients with idiopathic pulmonary hemosiderosis were lost to follow-up, and the other cases did not develop hemoptysis again. CONCLUSIONS: Idiopathic pulmonary hemosiderosis is the main cause of recurrent hemoptysis in children. Contrast-enhanced lung CT scans and pulmonary and bronchial arteriography are important methods for the etiological diagnosis of recurrent hemoptysis in children. Treatment methods should be selected according to the etiology.

[Changes in IL-17 and TGF-ß1 levels in serum and bronchoalveolar lavage fluid and their clinical significance among children with asthma].

OBJECTIVE: To investigate the changes in the levels of interleukin-17 (IL-17) and transforming growth factor beta 1 (TGF-ß1) in serum and bronchoalveolar lavage fluid (BALF) and their clinical significance among children with asthma. METHODS: Fifty-six children with asthma were divided into moderate or severe asthma (n=37) and mild asthma groups (n=19) and 18 children without asthma were selected as the control group. Cells in BALF were counted under a microscope. The levels of IL-17 and TGF-ß1 in serum and BALF were measured using ELISA. RESULTS: were no significant differences in total cell count and percentage of macrophages between the two asthma groups and the control group (P>0.05). The percentages of neutrophils, eosinophils and epithelial cells in BALF were significantly higher in the two asthma groups than in the control group (P<0.05). The two asthma groups had significantly higher levels of IL-17 and TGF-ß1 in serum and BALF than the control group (P<0.05), and the moderate or severe asthma group had significantly higher levels of IL-17 and TGF-ß1 in serum and BALF than the mild asthma group (P<0.05). Levels of IL-17 and TGF-ß1 in serum were significantly positively correlated with those in BALF (r=0.935 and 0.943, P<0.05 for both). In children with asthma, serum IL-17 level was significantly positively correlated with the percentage of neutrophils, eosinophils and epithelial cells in BALF (r=0.802, 0.799, and 0.674, P<0.05 for all), and a significant positive correlation was also seen between serum levels of IL-17 and TGF-ß1 (r=0.878, P<0.05). CONCLUSIONS: Levels of IL-17 and TGF-ß1 in serum and BALF are elevated in children with asthma. IL-17 and TGF-ß1 may be involved in the occurrence and development of asthma, and they play important roles in asthma attack and aggravation.

Dexamethasone reduces IL-17 and Tim-3 expression in BALF of asthmatic mice.

This study investigated the expression of interleukin-17 (IL-17) and T cell immunoglobulin mucin and domain-containing molecule-3 (Tim-3) in bronchoalveolar lavage fluid (BALF) of asthmatic mice and the effect of dexamethasone (DEX) on these factors. Thirty-six mice were randomly divided into three groups: normal group, asthmatic group and DEX group. The mouse model of asthma was established by sensitization with ovalbumin in both the asthmatic and DEX groups. The levels of IL-6, IL-10, IL-17 and TGF-ß were measured in BALF by enzyme-linked immunesorbent assay (ELISA). The mRNA expression level of Tim-3 was detected by reverse transcription polymerase chain reaction (RT-PCR). The ratio of Tim-3+CD4+ cells to total CD4+ cells in BALF was determined by flow cytometry. Differential inflammatory cells in BALF were detected. The correlations among IL-17, IL-6, IL-10, Tim-3 and inflammatory cells were analyzed. The results showed that the levels of IL-17, IL-6 and Tim-3 were substantially increased and the IL-10 level decreased in BALF in the asthmatic mice, which was significantly reversed by DEX treatment. IL-17 expression was positively correlated with IL-6 and Tim-3 expression and the number of inflammatory cells but negatively with IL-10 expression. These results indicate that the increased expression of IL-17 and Tim-3 in BALF may be implicated in the occurrence and development of asthmatic inflammation; the mechanism by which DEX suppresses asthmatic airway inflammation involves down-regulation of IL-17 and Tim-3 levels.

Small interfering RNA targeting T-cell Ig mucin-3 decreases allergic airway inflammation and hyperresponsiveness.

Since CD4+ T cells play a pivotal role in the development of airway inflammation and hyperresponsiveness, targeting activated CD4+ T cell subsets and increasing the cells with regulatory function would be a logical therapeutic approach. We showed that this outcome can be achieved by local therapy with Tim-3, which is a negative regulator of CD4+ T cells. Tim-3 expression was up-regulated by ovalbumin (OVA) induction. Attenuating Tim-3 expression by RNA interference suppressed allergen-induced immune responses. Intranasal application of Tim-3 shRNA diminished airway inflammation and hyperresponsiveness. Multiple mechanisms were involved in the inhibitory effects, including regulation the imbalance of Th1/Th17 and increasing Treg cell expression. Our results indicate that the Tim-3 pathway is highly involved in the regulation of asthma. Targeting Tim-3 by siRNA may hold therapeutic potential in preventing the development of allergic asthma.

Changes in interleukin-17 and transforming growth factor beta 1 levels in serum and bronchoalveolar lavage fluid and their clinical significance among children with asthma.

OBJECTIVES: To investigate the changes in the levels of interleukin-17 (IL-17) and transforming growth factor beta 1 (TGF-ß1) in serum and bronchoalveolar lavage fluid (BALF) and their clinical significance among children with asthma. METHODS: A total of 56 children with asthma were divided into moderate or severe asthma (n=37) and mild asthma groups (n=19) and 18 children without asthma were selected as the control group. Cells in BALF were counted under a microscope. The levels of IL-17 and TGF-ß1 in serum and BALF were measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: There were no significant differences in total cell count and percentage of macrophages between the two asthma groups and the control group (P>0.05). The percentages of neutrophils, eosinophils and epithelial cells in BALF were significantly higher in the two asthma groups than in the control group (P<0.05). The two asthma groups had significantly higher levels of IL-17 and TGF-ß1 in serum and BALF than the control group (P<0.05), and the moderate or severe asthma group had significantly higher levels of IL-17 and TGF-ß1 in serum and BALF than in the mild asthma group (P<0.05). The serum levels of IL-17 and TGF-ß1 were positively correlated with those in BALF (r=0.935 and 0.943, both P<0.05). In children with asthma, serum IL-17 level was positively correlated with the percentages of neutrophils, eosinophils, and epithelial cells in BALF (r=0.802, 0.799, and 0.674, all P<0.05), and a positive correlation was also seen between serum levels of IL-17 and TGF-ß1 (r=0.878, P<0.05). CONCLUSIONS: Levels of IL-17 and TGF-ß1 in serum and BALF are elevated in children with asthma. IL-17 and TGF-ß1 may be involved in the occurrence and development of asthma, and they play important roles in asthma attack and aggravation.