bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immune Sensing of Aeroallergen-Associated Double-Stranded RNA Triggers an IFN Response and Modulates Type 2 Lung Inflammation
Authors: Alanazi Hamad , She Li , Yan Liping , Sun Yilun , Zou Yi , Dube Peter , Brooks Edward , Barrera Gema , Lai Zhao , Chen Yidong , Liu Yong , zhang Xin , Li Xiao-Dong 
Published: January 2023

Abstract
Over 20 million Americans are affected by allergies such as asthma and allergic rhinitis, which can be caused by common allergens like house dust mites (HDM) or their associated components. The exacerbation of allergies can be fatal in patients with pre-existing chronic lung inflammation. Until now, there is no cure for asthma and type 2 allergies and the only available treatments aim to reduce the disease symptoms. Thus, there is an urgent need to understand the pathogenic mechanism of HDM-induced allergic responses. Since HDM play a major role in triggering allergic reactions, several studies focused on identifying their constituents. Although it is important to study HDM protein components that elicit adaptive type 2 inflammation (e.g. Der p 1), the role of innate sensors such as Toll-like receptors (TLRs) is unknown. A number of studies investigated presence of common adjuvants that may activate host immune receptors and modulate allergic diseases, for example, lipopolysaccharide (LPS) contaminating HDM triggered asthma in mice through acting on TLR4. In addition, single and double-stranded nucleic acids from bacteria or viruses can also act as adjuvants that stimulate innate immune receptors. Clearly, diverse allergen-associated components play an essential role in modulating type 2 lung inflammation in allergic patients. Recently, we have shown that HDM contain immunogenic RNA capable of modulating type 2 lung inflammation by inducing protective interferon responses. Using RNA-seq analysis, we show that HDM contain previously uncharacterized double stranded RNA sequences derived from novel RNA-viruses. Our findings suggest that other allergens may also have immunostimulatory RNA that can regulate type 2 lung inflammation.
IL-33 and allergen exposure promote development of “memory-like” ILC2s in the Airways
Authors: Ferrini Maria M , Jaffar Zeina , Roberts Kevan 
Published: January 2023

Abstract
The alarmin, IL-33 plays a critical role in the inception of lung inflammation and the exacerbation of allergic airway disease. Inhalation of environmental allergens and particulates is known to promote IL-33 release, and the cells responsive to this cytokine include group 2 innate lymphoid cells (ILC2s), dendritic cells and natural killer (NK) cells. In this study, we evaluated the effect of IL-33 and house dust mite allergen inhalation on the properties of lung ILC2s. Instillation of IL-33 into the airways of mice resulted in a marked elevation in number of ILC2s and eosinophils. Importantly, a notable feature of the inflammatory response was an increase in CD103 expression by ILC2 (from 3% to 45%). These CD103+ ILC2s appeared to represent memory-like ILC2s since they were highly responsive to IL-25 and produced significant levels of IL-9 and IL-13. Critically, inhalation of house dust mite allergen also increased CD103 expression by ILC2s. Given that CD103 expression can be induced by TGF-β, we next determined whether active TGF-β1 is released following IL-33-driven lung inflammation. Remarkably, the levels of active TGF-β1 present in the airways were markedly raised following IL-33 instillation, and this was responsible for a reduction in IFN-γ-producing NK cells and raised CD226 (DNAM-1) expression. Collectively, these findings reveal that IL-33 and allergen promote the accumulation of CD103+ memory-like ILC2s in the airways. A role for TGF-β in promoting CD103 expression while suppressing NK cell-mediated regulation of the inflammatory process is proposed.
The emerging roles of neutrophils and macrophages during experimental acute pyelonephritis
Authors: de Dios Ruiz Rosado Juan , Robledo-Avila Frank , Becknell Brian , Partida-Sanchez Santiago 
Published: January 2023

Abstract
Every year, pediatric urinary tract infections (UTI) account for 1.5 million clinician visits in the US alone. Up to 50% of infants with UTI develop a kidney infection (acute pyelonephritis, APN). Despite antibiotic therapy, 15% of children with APN develop renal scarring, which is associated with deterioration in kidney function. Here, we determined the contribution of macrophages and neutrophils to bacterial clearance and the development of kidney fibrosis during experimental APN.
Female C3H/HeOuJ mice were treated, via retro-orbital, with one of the following antibodies: isotype IgG (control), anti-Ly6G (depletes neutrophils), anti-CD115 (depletes monocytes), and anti-GR1 (depletes monocytes and neutrophils). After cell depletion, the animals were transurethrally infected with UPEC. Ascending UTI was evaluated via biophotonic imaging. The dynamics of monocytes and neutrophils were determined by flow cytometry. Histopathological scores were performed by H&E and Sirius-Red stainings. Transcriptomic analyses of kidneys were performed using TaqMan Arrays.
Our data indicate that neutrophils are required for bacterial clearance, while macrophages promote inflammation in the infected urinary tract. However, the absence of neutrophils results in increased macrophage infiltration and exaggerated macrophage-dependent inflammatory responses in the bladder and kidney. Also, our results demonstrate that macrophage-dependent inflammation during APN, leads to renal scarring and renal functional impairment. These findings uncover a role for the neutrophil-macrophage imbalance in promoting kidney fibrosis during APN. This knowledge will be helpful to develop novel therapies to prevent post-APN scarring in children.
Enhancement of vitamin C on PMN function in healthy individuals
Authors: Ruknarong Laongthip , Nonsa-ard Rujira , Chuangchot Nisa , Jumnainsong Amonrat , Leelayuwat Naruemon , Jusakul Apinya , Leelayuwat Chanvit 
Published: January 2023

Abstract
Vitamin C (ascorbic acid, ascorbate) promotes immune cell activities by supporting various cellular functions especially, in neutrophils or polymorphonuclear (PMN) leukocytes, which play an important role in bacterial infection and acute inflammation. Little data have been reported in impact of vitamin C on PMN function during bacterial phagocytosis in healthy individuals. Here, we have elucidated the effect of vitamin C on PMN phagocytosis and oxidative burst to Staphylococcus aureus by whole blood assay and flow cytometry after pre-incubation without and with vitamin C in vary concentrations. The results showed that vitamin C concentration at 20 mM induced a significant increase phagocytosis (p-value=0.03) and had trend to significant increase oxidative burst (p-value=0.06) while vitamin C concentration of 30 mM showed significant increase both phagocytosis and oxidative burst (p-value=0.04 and 0.02, respectively) at only 15 min of exposure to the bacteria. As a minimal concentration that can increase phagocytosis, we selected vitamin C at 20 mM as optimal concentration to investigate more data by enrollment of healthy volunteers (n=17). The results revealed significant difference both phagocytosis and oxidative burst after 20 mM vitamin C pre-incubation (p&lt;0.0001). In conclusion, we showed that vitamin C at 20 mM can boost phagocytosis and oxidative burst of neutrophils in healthy adults within 15 min after bacterial exposure. This rapid activation may be applied to fight bacterial infection. Nevertheless, further research in clinical trial and underlying mechanisms await further studies.
Identification of N-formylated peptides as the neutrophilic chemotactic factors of Mycobacterium tuberculosis
Authors: Dean Kristin T , Jung Bockgie , Samten Buka 
Published: January 2023

Abstract
Since tuberculosis (TB) remains a major cause of human death, it is urgent to understand the pathology of TB. Lung neutrophil infiltration initiates granuloma formation and is associated with disease severity. Though several cytokines and chemokines are associated with neutrophil infiltration in chronic inflammation, neutrophil chemotactic factors of Mycobacterium tuberculosis (Mtb) remain unexplored. Thus, we performed neutrophil chemotactic assays by culturing neutrophils of healthy human blood samples and mouse bone marrow in the upper chamber of a transwell plate with 5–8 μm pores with the culture filtrates of Mtb H37Rv (CF) in the lower chamber using N-formyl-met-leu-phe (fMLF), a bacterial neutrophil chemotactic tripeptide, as a positive control. The cell numbers in the lower chamber were determined by flow cytometry as a chemotactic indicator. Our results show that CF induced chemotaxis of both human and mouse neutrophils in an Mtb culture period dependent manner. Testing CF of 11 clinical isolates of Mtb also showed neutrophil chemotactic activity. Sulfasalazine, a formylated peptide receptor inhibitor, blocked chemotaxis of neutrophils, indicating the involvement of fPRs in CF induces chemotaxis of neutrophils. Mass spectrometry analysis of CF identified three candidate N-formylated heptapeptides. Authentication of the chemotactic activities of the identified peptides with synthetic mimetics confirmed their neutrophil chemotactic activities and indicated the requirement of N-formylation for their chemotactic activities. Thus, we have identified novel formylated peptides of Mtb with neutrophil chemotactic activities, and further studies are in progress to determine their significance in TB infection.
The TRPM2 ion channel regulates the inflammatory response of neutrophils during Listeria monocytogenes infection
Authors: Robledo-Avila Frank Hanz , de Dios Ruiz Rosado Juan , Brockman Kenneth L , Partida-Sanchez Santiago 
Published: January 2023

Abstract
Calcium signaling is required for most antimicrobial functions of neutrophils. The transient receptor potential melastatin-2 (TRPM2) cation channel has been proposed to play important roles in modulating calcium mobilization, cell migration and oxidative stress in neutrophils. However, the precise mechanism by which TRPM2 activation may exert a pro- or anti-inflammatory function remains unclear. In this study, we used a mouse model of Listeria monocytogenes infection to define the role of TRPM2 in the regulation of neutrophils’ function. We found that infected Trpm2−/− mice were associated with high bacterial burden in spleen and liver indicating increased susceptibility to Listeria infection. Pronounced migration rates of neutrophils and monocytes to the liver and spleen characterized the inflammatory response to Listeria during the first 18h. In the Trpm2−/− mice, the acute phase infection resulted in septic shock, defined by increased serum levels of TNF-α, IL-6, and IL-10, but no decreased levels of IFN-γ or IL-12. Furthermore, depletion of neutrophils demonstrated the critical role of these cells in regulating acute inflammation by conferring resistance of Trpm2−/− mice to Listeria infection. Gene expression and inflammatory cytokine analyses of infected tissues further confirmed the hyperinflammatory profile of Trpm2−/− neutrophils. Finally, the increased inflammatory properties of Trpm2−/− neutrophils correlated with cellular calcium overloading and potentiated membrane depolarization, in response to Listeria infection. In conclusion, our findings suggest that TRPM2 channel plays critical roles in the regulation of the inflammatory properties of neutrophils during Listeria monocytogenes infection.
Role of invariant nature killer T cell in dysbiosis of gut microbiota among Mycobacterium tuberculosis infected patients
Authors: Huang Shiang-Fen , Su Wei-Juin 
Published: January 2023

Abstract

Background
In Mycobacterium tuberculosis (MTB)-infected patients, the dysbiosis in gut microbiota was associated with systemic proinflammation. The invariant nature killer T (iNKT) cell was suggested the T-cell involved in immune modulation activated by metabolites or structural molecules of microbial in gut lumen. Herein we investigated how the gut microbiota influence on the systemic immunity by study the iNKT cells in peripheral blood in human.

Method
Patients with active TB, latent TB infection (LTBI), and healthy controls (HC) were enrolled. Interleukin (IL)-10, IL-4, IL-6, and IL-1B levels, the CD3+, CD4+, CD8+ cells, iNKT (CD3+6B11+) cells counts in blood, proliferation index (PI) of iNKT cells by α-galactosylceramide (α-GalCer) stimulation in vitro, and the interferon-gamma (IFN-γ) concentration by IFN-r-releasing-assay (IGRA) were measured.

Results
In active TB, both the cell counts and PI of iNKT were decreased. In LTBI, the PI of iNKT cells and the stimulated IFN-γ by MTB-Antigen divided to the iNKT cell counts (IFN-TBAg/iNKT) were positively associated with the relative abundance of Firmicutes. In active TB, a higher iNKT cell counts were also associated with higher relative abundance of Firmicutes. After adjusted by iNTK cell counts, the proinflammatory cytokines IL-10, IL-1B, and IL-6 were positively associated with the relative abundance of Bacteroidaceae in the gut microbiota.

Conclusion
In LTBI, Firmicutes in the gut microbiota determined the activities of iNKT cell and the IFN-TBAg level in peripheral blood. In active TB, the iNKT cell was related to the pro-inflammation and the relative abundances of Bacteroidaceae in gut microbiota.
CALCIUM MOBILIZATION AND EXPRESION OF TRPM2 AND CRAC CHANNELS ARE ALTERED IN HUMAN NEUTROPHILS WITH AGING
Authors: Sumoza-Toledo Adriana , de los Ángeles Vázquez-Prieto María , Lascurais-Santamaría Nallely , Fernández-Eufrasio Nilda Belén , Montiel-Condado Dvorak , Garibay-Escobar Adriana 
Published: January 2023

Abstract
Elderly susceptibility to infectious diseases may be a result of a decline in the function of neutrophils. We analyzed the number and percentage of neutrophils in peripheral blood of healthy elderly (60–85 years old) and young adults (20–35 years old), as well as the effect of aging on store-operated Ca+2 entry (SOCE), Ca+2 mobilization towards 100 nM or 1μM fMLP, 300 nM IL-8, 10 nM C5a, and 25 μg/ml LPS, by flow cytometry; and its relation to the expression of CD88, CXCR2, FPLR1, TRL4 and TRPM2 and CRAC channels. To examine Ca+2 mobilization, cells were loaded with 1 μM Fluo-4-AM. Baseline Ca+2 level was recorded for 60 s before the addition of the stimulus; response was recorded up to 10 min. Subcellular expression of CRAC and TRPM2 was investigated by confocal microscopy and their mRNA quantified by real time RT-PCR. Finally, migration towards 10 nM fMLP, C5a or IL-8 was measured by transwell chemotaxis assay. The results showed increased numbers and percentage of neutrophils in elderly, mainly in females. However, these cells presented lower Ca+2 entry in response to C5a, IL-8 and fMLP, and significantly lower SOCE. We did not find difference in the Ca+2 response to LPS or in the expression of the receptors of the stimuli (CD88, CXCR2, FPLR1 and TRL4). A punctuated intracellular pattern of TRPM2 and CRAC was observed in the neutrophils. TRPM2 was located in late endosomes and presented changes in its distribution with age. It was also detected a decrease in the mRNA of the TRPM2 and CRAC channels, mainly in women. Finally, we found that neutrophils from elderly migrated less towards fMLP, IL-8 and C5a. Our data suggest that the mechanisms that underlie the decrease on neutrophil functions with aging might be alterations in Ca+2 signals.
Human influenza-specific adaptive NK cells mediate potent HLA-E-restricted responses against antigenically distinct influenza strains
Authors: Jost Stephanie , Lucar Olivier , Yoder Taylor , Werner Alexandra , Ghofrani Joshua , Reeves R. Keith 
Published: January 2023

Abstract
Novel influenza vaccine approaches to achieve cross-protection against a broad range of antigenically distinct strains are needed. Beyond their ability to eliminate infected cells without the need for prior sensitization, natural killer (NK) cells have been shown to mediate virus-specific recall responses. Notably, exposure to influenza antigens induces protective influenza-specific memory NK cells in mice, but evidence of influenza-specific memory NK cells in humans is still lacking.
Using intracellular cytokine staining, we found that NK cells from 45% of adults can robustly respond to nucleoprotein (NP) from at least 2 heterosubtypic influenza strains completely independent of T cell help. To further determine if a subset of NK cells can specifically react to NP peptide pools from distinct strains, we clonally expanded individual NK cells (NKCL) and measured H1N1, H2N2 and H3N2 NP-specific killing by 33 NKCL from 6 donors. 63% of NKCL showed positive responses against the 2009 H1N1 pandemic strain (2%–57% specific lysis). Strikingly, half of H1N1 NP-specific NKCL also displayed cytotoxicity against either H2N2 or H3N2 (9%–100% specific lysis) or against both H2N2 and H3N2 strains (5%–11% specific lysis). Potent killing by NKCLs was associated with high cell surface expression of the activating NKG2C receptor, a ligand for HLA-E. Accordingly, we identified an HLA-E-restricted NP epitope that triggers activation of influenza-specific NK cells.
Collectively, these data provide the first mechanistic evidence of influenza-specific memory NK cells in humans and suggest influenza-specific responses mediated by NK cells may have the potential to be harnessed to enhance influenza vaccine-induced cross-protection.
Role of CD34+CD10+CD19−Pax5+ cells on M2b macrophage polarization
Authors: OHAMA Hideko , Asai Akira , Yasuoka Hidetaka , Tsuchimoto Yusuke , Fukunishi Shinya , Higuchi Kazuhide 
Published: January 2023

Abstract
M2b macrophages (Mf)/monocytes play a role in the increased susceptibility of alcoholics to infections. However, the mechanism involved in M2bMf polarization remains unclear. In this study, we investigated the role of CD34+CD10+CD19−Pax5+ cells (Pre/Pro-B cells) in the peripheral blood of alcoholics on alcohol-related M2bMf/monocyte polarization through the production of high mobility group box-1 (HMGB1). M2b monocytes were identified as few bactericidal activity, IL-10 and CCL1 productions. Pre/Pro-B cells isolated from 18 alcoholics and 11 healthy donors (HD) were cultured with 10 nM of retinoic acid for 24hrs and culture medium obtained were assayed for HMGB1 by ELISA. CD14+cells (1 × 106 cells/ml) from HD were stimulated with 10% (v/v) of each culture media of Pre/Pro-B cells for 24hrs and their producing abilities of IL-10 and CCL1 were assayed. And 1 × 106 cells/ml of these cells were infected with 2 × 106 CFU /ml Enterococcus faecalis (E. faecalis). The bactericidal activity was assayed for colony counting methods. In the results, Pre/Pro-B cells from HD did not produce HMGB1(10.5 ± 5.5 ng/mL), while these cells from alcoholics did (70.3 ± 20.2 ng/mL). When CD14+ cells from HD were stimulated with each culture media from alcoholics, these CD14+ cells produced IL-10 (102.3 ± 1.7 pg/mL) and CCL1 (3.12 ± 1.33 ng/mL) and did not have bactericidal activity against E. faecalis (17.0 ± 8.6%). However, same CD14+ cells from HD were stimulated with each culture media from HD, these cells did not produce IL-10 (10.3 ± 3.4 pg/mL) and CCL1 (0.3 ± 0.01 ng/mL) and have the bactericidal activity (62.8 ± 3.8%). These results indicate that Pre/Pro-B cells play a crucial role in M2bMf/monocyte polarization via HMGB1 production in alcoholics.
Type III interferons are expressed in tuberculosis granulomas and promote an inflammatory phenotype in macrophages that differs from type I interferon
Authors: Talukdar Priyanka , Mattila Joshua T. , Fallert Junecko Beth A 
Published: January 2023

Abstract
Humans and non-human primates express four subtypes of type III interferons (IFNλs; IFNλ1-IFNλ4). Unlike type I interferons, which have been thoroughly investigated in tuberculosis (TB), the role of IFNλs and their effects on immunity in TB remain unknown, especially at the disease site. To better understand the role of IFNλs in TB, we examined IFNλ1 and IFNλ4 expression in cynomolgus macaques with TB and investigated the effects of IFNα/β, IFNλ1 and IFNλ4 signaling on macaque macrophages. Using IHC, we identified differential IFNλ1 and IFNλ4 expression in macrophages and neutrophils, including IFNλ4 localization in the nuclei of epithelioid macrophages. CD206+ macrophages and CD3+ cells in the airway expressed IFNλR while CD11c+ macrophages in the lymphocyte cuff of the granuloma expressed IFNλR. To measure IFNλ1 and IFNλ4’s effect on macrophage gene expression and compare these cytokines against IFNα/β, we used NanoString transcriptional profiling and Ingenuity Pathway Analysis (IPA) on cytokine-stimulated macrophages to identify differentially regulated pathways. We found that IFNα/β upregulated the greatest number of ISGs, followed by IFNλ1, whereas IFNλ4 stimulation had minimal effect on gene expression. Interestingly, pro-inflammatory genes including IL-1β, IL-8, TLR1, BATF were upregulated by IFNλ1 while IL-1β and TLR1 were downregulated by IFNα/β. To determine the effects of IFNλs on anti-mycobacterial macrophage responses, we used a reporter Mtb strain to determine how IFNλ1 and IFNλ4 effect the viability of Mycobacterium tuberculosis. Our data suggest that IFNλs have non-redundant properties with type 1 interferons that may promote macrophage activation, inflammation, and antibacterial activity in TB.
Effect of temperature on functional activity of macrophages in three different species
Authors: Hassan Intisar A , Renfro Zachary , Blake Harrison , Rath Satyajit , Durdik Jeannine M 
Published: January 2023

Abstract
Temperature affects body physiological functions, including immunity in ways that influence survival. This study investigated the effect of temperature on macrophage activation and metabolism across species. Macrophages were isolated from mice, chicken or rainbow trout primary tissue or from macrophage cell lines and were activated with lipopolysaccharide (LPS) or IFN-γ. For mice, both Raw264.7 and primary cells, nitric oxide (NO) production was similar at 35°C and 37°C but dropped dramatically at temperatures below 35°C. At fever temperature (39°C), NO release increased in response to LPS. Young bone marrow derived macrophage (BMDM) and peritoneal resident macrophage (PRM) showed increased protein synthesis at 39°C compared to 37°C. Chicken splenic macrophages (CSM) showed NO responses that were similar at 37, 39, and at 41°C (normal for avian). A fever of 42°C had a large stimulatory effect on NO production compared to 41°C. A chicken liver derived macrophage cell line, HTC, showed the same pattern. They also showed higher protein synthesis at 42°C compared with (41°C) after LPS stimulation. Trout head-kidney macrophages (THM) showed the highest NO responses at 19°C when compared to more typical stream temperatures of 13, 15, and 16°C. They still showed some response at 28 and 37°C. Thus, their macrophages respond at higher temperatures than the fish can tolerate. Stimulated THM cells with LPS at 19°C showed increased protein synthesis compared to 13 and 16°C. We can conclude from our experiments that fish macrophages had a much broader range of temperatures at which they could respond by NO generation and protein synthesis compared to mice and chickens and all had increased responses at fever temperatures.