bluuebunny/crossref_metadata_2025_split huggingface dataset:
Eosinophil-derived IL-4 drives inflammatory dilated cardiomyopathy
Authors: Diny Nicola , Barin Jobert G , Talor Monica , Rose Noel R , Cihakova Daniela 
Published: January 2023

Abstract
Inflammatory dilated cardiomyopathy (DCM) is a major cause of heart failure in children and young adults. DCM develops in up to 30% of myocarditis patients, but the mechanisms involved in disease progression are poorly understood. Patients with eosinophilia frequently develop cardiomyopathies. We determined the role of eosinophils in myocarditis and DCM using the experimental autoimmune myocarditis (EAM) model. In wildtype (WT) BALB/c mice, EAM progresses to DCM and heart failure. Eosinophil-deficient ΔdblGATA1 mice developed myocarditis but showed no signs of heart failure. Thus, eosinophils were dispensable for myocarditis induction but required for progression to DCM. EAM in hypereosinophilic IL5Tg mice resulted in eosinophilic myocarditis with severe atrial inflammation and progressed to severe DCM. Atrial inflammation was also observed in 2 of 3 patients with eosinophilic myocarditis. The DCM phenotype required eosinophils but not IL5. IL5TgΔdblGATA1 mice were protected from DCM while IL5−/− mice exhibited DCM comparable to WT mice. Eosinophils drove progression to DCM through their production of IL4. In our experiments, eosinophils were the major IL4 expressing cell type in the heart during EAM. Like ΔdblGATA1 mice, IL4−/− mice were protected from DCM and eosinophil-specific IL4 deletion resulted in improved heart function. In conclusion, eosinophils drive progression of myocarditis to DCM, cause severe DCM when present in large numbers, and mediate this process through IL4.
Modulation of human innate lymphoid cell function by IL-10
Authors: Bonne-Annee Sandra , Nutman Thomas B 
Published: January 2023

Abstract
Although present in extremely low numbers in the human circulation at steady state, innate lymphoid cells (ILCs) are expanded in human helminth infections. Despite their low frequency (accounting for 0.21% (range 0.023–3.58%) of lymphocytes in the peripheral blood under homeostatic conditions), ILCs release extremely large quantities of cytokines following activation. If left unchecked, these cytokines can have deleterious effects on the host. We have observed that circulating ILCs demonstrate surface expression of IL-10R under homeostatic conditions. To examine how the expression of IL-10R regulates cytokine production by ILCs, ILC subsets were isolated from the peripheral blood of healthy donors by flow cytometry-based sorting and activated in the presence or absence of the immunoregulatory cytokine IL-10. ILCs stimulated in the presence of IL-10 had a marked reduction (range from 2–3.6 fold) in cytokines (IL-4, IL-5, IL-13, IL-17A, INF-γ, and TNF-α) when compared to ILCs activated without IL-10. These findings demonstrate that IL-10 signaling can inhibit cytokine production by activated ILCs. Furthermore, studies are currently underway to examine ILC subsets in the context of chronic helminth infections, where IL-10 has a modulatory effect on innate and adaptive immune cells and to identify additional mechanisms that regulate ILC cytokine production, including TGF-β.
Characterization of an alternatively activated IL-10 producing innate lymphoid type-2 effector cell population
Authors: Kaye Jonathan , Seehus Corey Ray , Kadavallore Asha , de la Torre Brian , Yeckes Alyson , Wang Yizhou , Tang Jie 
Published: January 2023

Abstract
Type-2 innate lymphoid (ILC2) cells share cytokine and transcription factor expression with CD4+ Th2 cells, but it is unknown if this ILC subtype undergoes functional diversification. Here, we report in vivo induction of a molecularly distinct subset of activated lung ILC2 cells, termed ILC210 cells, that produce IL-10 and downregulate genes associated with inflammation. Signals that generate ILC210 cells were distinct from those that induced IL-13 production and, along with transcriptome data, suggested an alternative pathway of activation. In vivo, IL-2 greatly enhanced ILC210 cell generation by IL-33 and was associated with decreased eosinophil recruitment to the lung. Unlike the majority of activated ILC2 cells, the ILC210 cell population underwent contraction following cessation of IL-33 stimulation in vivo, but with maintenance of a subpopulation of cells that can be recalled by restimulation, analogous to T cell effector cell and memory cell generation. Together, these data demonstrate previously unappreciated heterogeneity in the ILC2 cell response.
Macrophage heterogeneity in human macrophages correlates with autophagy-dependent anti-mycobacterial activity and ability to present antigen to CD4 T cells
Authors: Khan Arshad , Mann Lovepreet , Papanna Ramesha , Won Jong Hak , Mani Arunmani , Canaday David , Jagannath Chinnaswamy 
Published: January 2023

Abstract
Tuberculosis due to Mycobacterium tuberculosis (Mtb) kills around 1.5 million people each year. Although only 10% of humans develop active disease after aerosol infection, the basis for differential susceptibility of humans remains unclear. To determine if human macrophages are heterogeneous in their ability to kill Mtb, cord blood derived macrophages were treated with either IFN-γ or IL-4 to prepare M1 and M2 phenotype in vitro. Human THP-1 macrophages were also similarly prepared. Macrophages were infected with a 1:1 ratio of Mtb, incubated and on days 3 and 5 post infection, macrophage lysates were plated for CFU counts. In addition, intracellular autophagy was measured in gfpMtb infected macrophages using cytoID stains to detect autophagosomes and LC3 lipidation through western blot of macrophage lysates. Finally, Mtb infected macrophages were overlaid using CD4 T cells specific for the Antigen-85B epitope to measure antigen presentation. M1 macrophages showed a bacteriostatic ability and restricted the growth of Mtb while IL-4 treated M2 macrophages showed nearly 1 log increase in the growth of Mtb. Thus M2 macrophages were permissive for growth of Mtb. Consistent with this observation, gfpMtb co-localized with autophagosomes more frequently in M1 macrophages suggesting a role for autophagy during growth inhibition. LC3 lipidation was better in M1 compared to M2 macrophages. Finally, M1 macrophages presented Ag85B to CD4 T cells better than M2 macrophages. These observations suggest that macrophage heterogeneity into M1 and M2 is associated with an innate ability to kill intracellular tuberculosis and trigger immune activation.
TLT-1 as a possible modulator of Systemic Lupus Erythematosus.
Authors: Morales Jessica , Rodriguez Yerania , Vila Karina , Ricaurte Martha , Vila Luis M , Washington Anthony V. 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by immune system hyperactivity and autoantibody production that can affect almost any organ system. TREM like transcript-1 (TLT-1) is a membrane receptor found in platelet α-granules with a soluble form (sTLT-1) detected in serum but not plasma of healthy individuals. The evaluations of patients suffering from diseases, such as sepsis, show a significant increase of plasma sTLT-1 levels. It is not known if TLT-1 plays a role in auto immune diseases such as Lupus. In the present study we evaluate the potential that TLT-1 may play a role in the etiology of SLE. We assessed the plasma of 46 SLE patients and compared to 28 healthy controls by ELISA. Our results showed that SLE patients had a significant decrease in plasma sTLT-1 levels compared to healthy controls (9.0 [7.2] vs. 18.6 [22.3] ng/ml, p=0.008). A negative correlation was observed between TLT-1 levels and LupusPRO [lupus symptoms (r = −0.388, p = 0.055). Based on this data; we hypothesized that SLE patients produce autoantibodies against TLT-1 in serum which might correlate with clinical aspects of the disease. To address this question; TLT-1 antibodies levels in serum were measured by ELISA. Our data suggests that a sub-population of SLE patients have TLT-1 autoantibodies. This data was further confirmed by confocal microscopy. Using HEK cells stably transfected with TLT-1; we evaluated the serum of SLE patients for the presence of TLT-1 autoantibodies using immunofluorescence. Our data confirms that a sub-population of SLE patients with active disease expresses antibodies against TLT-1 suggesting a possible role for TLT-1 during the pathogenesis of SLE and targets new treatment possibilities for lupus patients.
Cellular gatekeepers of the humoral immune response
Authors: Yousif Ashraf S , Omatsu Tatsushi , Weaver Grant , Shah Pankaj , Reinnecker Hans-Christian , Lingwood Daniel 
Published: January 2023

Abstract
The spleen is critical for eliminating bloodborne pathogens and preventing systemic infection. In the marginal zone (MZ), the site in which splenic antibody responses are first initiated, we have discovered a dendritic cell (DC) ‘hairnet’ architecture that is responsible for extracting two unrelated viral antigens from the perfusing blood and importing them into B cell follicle to initiate IgM and IgG responses. The IgM response was independent of T-cell help, and proceeded via DC display of conformationally intact antigen in the absence of MHC proteins. Interestingly the DC-antigen captures and importing mechanism are solely governed by the glycan of the antigens. Taking all together, we propose that DC hairnet architecture, inshealthing the MZ, constitutes a filtering device that capture and concentrates protein antigens in a glycan-dependent manner from solution and catalyzes their delivery to B cells follicle to initiate humoral immune responses.
Human myeloid derived suppressor cell (MDSC) subset phenotypes
Authors: Talmadge James E , Cole Kathryn , Britton Holly , Dafferner Alicia , Warkentin Phyllis 
Published: January 2023

Abstract
MDSCs are increased in cancer patients and were originally defined based on T-cell suppression and a negative correlation with prognosis and overall survival and a direct correlation with tumor stage. They were originally described, based on CD34 expression and a hematopoietic progenitor phenotype. However, their phenotypic definition is investigator dependent in association with suppressor cell function. There are two main subsets defined based on a lack of lymphoid and differential expression of myeloid markers resulting in a cellular continuum from multi-potential hematopoietic progenitors to differentiated granulocytes and monocytes. One recent hypothesis is that MDSCs form three subsets including macrophage MDSC (CD14+ and no CD15 or CD66b); granulocytic MDSCs (CD15 and/or CD66b expression and no CD14); and immature (i-) MDSC (linage (Lin) negative, CD33+ cells). Unfortunately, there is no consensus on markers extent to CD14 and CD15 with linage and HLA-DR negativity providing a frequent; although not universal theme. We report a flow cytometry gating strategy utilizing a single staining tube and a linear phenotypic characterization of MDSC subsets supported by density and T-cell suppression. We suggest that staining with antibodies to Lin, HLA-DR, CD14, CD15, CD11b and CD33 is sufficient to subset into the three phenotypes that can be further subset based on CD16. PDL1 and CD34 expression. The majority of iMDSC are CD34+CD33+ cells supporting a plastic phenotype and myeloid commitment and can be further, delineated based on CD16 expression supporting immaturity. We note, that some cancer patients express MDSCs at high levels; although the subsets preponderance varies between patients.
Scavenger receptor CD36 on B cells senses modified self-antigens to prevent autoimmunity
Authors: Duhlin Amanda , Grasset Emilie K , He Chenfei , Amara Khaled , Sippl Natalie , Lindh Emma , Vargas Leonardo , Dahlberg Carin , Westerberg Lisa , Smith Edvard CI , Malmström Vivianne , Pierce Susan K , Karlsson Mikael C 
Published: January 2023

Abstract
Scavenger receptor CD36 has been shown to mediate uptake of modified self-antigens such as apoptotic cells and oxidized LDL in macrophages and dendritic cells. Interestingly, CD36 was discovered to be preferentially expressed also on an innate B cell subtype, marginal zone B cells (MZB). Here we investigate the role of CD36 in B cell activation in the context of apoptotic cell clearance and break of tolerance to self. We found that formation of germinal center B cells and autoantibody production in response to an increased load of apoptotic cells coincide with downregulation of CD36 on MZB. We could also in connection to this show that systemic lupus erythematosus (SLE) patients have lower levels of peripheral MZB compared to healthy individuals. B cells lacking CD36 proved more easily activated in response to apoptotic cells, as was shown in bone marrow (BM) chimeras reconstituted with wild type and CD36-deficient BM. In steady-state, we found that CD36 co-localizes with the B cell receptor and the tyrosine kinase Lyn. Interestingly, upon cross-linking of the inhibitory Fc receptor, FcgRIIb, CD36 instead co-localizes with this receptor. This suggests that CD36 interacts with FcgRIIb to activate a negative signaling cascade in B cells. Our data implicate CD36 in the early regulation of B cell responses towards apoptotic cells and autoantibody production. In line with this we find that CD36-deficient B cells are more resistant to FcgRIIb-induced cell death. Thus, CD36 exerts its regulatory function by associating with a known negative signaling pathway in B cells involving Lyn and FcgRIIb. This finding could have implications for autoimmune diseases involving apoptotic cell-derived self-antigens, such as SLE.
IL-12 stimulates CTLs to secrete exosomes with biological functions
Authors: Xiao Zhengguo , Wang Yan , Li Lei 
Published: January 2023

Abstract
An effective CTL response is generally accomplished by massive CTL expansion and production of effector molecules, potentially exhausting resources and rendering the host vulnerable to other infections. It is not yet well known how the host maintains relative functional immunity against other pathogens during the course of an ongoing, robust CTL response against an active infection. We found that CTLs secrete exosomes in the presence and absence of IL-12, which all contain typical exosomal markers, such as ALIX and TSG101, as well as CTL proteins ZAP70 and granzyme B (GZB). However, when conditioned with IL-12, CTLs release smaller exosomes, which contain more proteins of exosomal markers, compared with exosomes secreted by control CTLs. More importantly, IL-12-conditioned exosomes can directly stimulate naïve CTLs to produce IFNγ, TNFα and GZB, which accompanied by upregulation of CD25, CD69 and CD44, independent of antigen or cytokines, whereas exosomes from control CTLs have minimal effects. In addition, IL-12-conditioned exosomes are able to strengthen weak TCR stimulation in CTLs. Proteomic analysis demonstrates that IL-12 stimulation skews exosomal proteins toward certain metabolism-related enzyme activities, including hydrolase, oxidoreductase and transferase, which are associated with CTL functions. Our findings indicate that the biological function and morphology of exosomes secreted by CTLs are influenced by the stimulation CTLs have received. This suggests that a functional, antigen-specific CTL response may boost functions of CTLs against other pathogens through secretion of functional exosomes. This may be a new mechanism for homeostatic regulation in CTL immune responses.
Ten-day exposure to low doses of ultrafine black carbon induces stress responses and reduces the vitality of human and murine pulmonary cell lines
Authors: Salinas M Esther 
Published: January 2023

Abstract
Ultrafine particulates (UFP) are emitted into urban air by combustion processes, commonly via vehicular emissions. These respirable particles can induce a series of immune responses contributing to higher incidences of allergy-related asthma. It is unclear how cells that are in direct contact with these inhaled materials are affected. Our project aims to elucidate the effects of ultrafine carbonaceous particle exposure to human and murine pulmonary cells. We surmise that prolonged treatments of ultrafine black carbon (UBC) directly contribute to lung irritation. In vitro studies were conducted using UBC treatments after a 10-day incubation period. Viability data has shown that the different concentrations of UBC used were mostly non-toxic and generally altered cellular function displaying an inverse dose-dependent manner. Biochemical assays indicated decreased metabolic activity; no significant negative effect on viability was observed in the epithelial cell lines. In the macrophages/monocytes, no damaging physiological activity and no compromising effects on cellular vitality were displayed. The induction of oxidative stress was also assessed using confocal microscopy showing a high number of ROS with increasing UBC concentrations, yet no marked trends were noticed upon conducting mitochondrial de-polarization assays via flow cytometry. This regimen demonstrates that various concentrations of UBC treatments affect mammalian cell function distinctly. In summation, this study suggests that carbonaceous pollutants negatively impact intracellular functionality and overall cellular health. Our findings may potentially provide treatments for allergy-induced asthma upon chronic exposure to ambient nanocarbon.
Gimap5-dependent inactivation of GSK3 is required for CD4 T cell homeostasis and prevention of immune pathology
Authors: Hoebe Kasper , Patterson Andrew , Endale Mehari , Lampe Kristin , Bleesing Jack 
Published: January 2023

Abstract
The GTPase of immunity-associated protein 5 (Gimap5) has been linked with lymphocyte survival, autoimmunity and colitis development. Previous studies indicated that Gimap5-deficient mice exhibit a reduced lymphocyte proliferation and survival. Here we show that Gimap5 is essential for inactivation of Glycogen Synthase Kinase-3β (GSK3β) following T cell activation, a constitutive active Ser/Thr kinase whose activity reduces β-catenin and c-Myc levels thereby maintaining T cell quiescence. Gimap5 regulates the subcellular localization of GSK3β and its phosphorylation at Ser389, which limits DNA damage during T cell proliferation. Importantly, pharmacologic and genetic targeting of GSK3 in mice can override Gimap5-deficiency in CD4+ T cells, preventing exacerbated DNA damage while enhancing T cell proliferation and survival. Additionally, this ameliorates immune-mediated pathology in vivo. Finally, we show that a human patient carrying a GIMAP5 loss-of-function mutation exhibits similar T cell defects and pathologies and exhibit a reduced T cell proliferation and survival associated with increased DNA damage that can be corrected in vitro with GSK3 inhibitors. T cells require a robust clonal expansion during an effective immune response. Given the predominant expression of GIMAP5 in lymphocytes, we propose that the identified pathway presents a novel and critical molecular mechanism required for productive CD4+ T cell responses and immune homeostasis in the periphery.
Lack of polyfunctional cytomegalovirus-specific T cells in hemodialysis patients
Authors: Chiu Yen-Ling , Lay Fang-Yun , Shu Kai-Hsiang , Chou Tzu-Ying , Chia Jean-San , Schneck Jonathan P 
Published: January 2023

Abstract
Polyfunctional T cells are critical for maintaining protection against pathogens. Patients with end-stage renal disease (ESRD) are at increased risks for infectious complications and their T cell immunity against viruses may be impaired. The current study intends to investigate the effects of renal disease on T cell immunity by analyzing T cell differentiation (based on expression of CCR7 and CD45RA) and polyfunctionality response against cytomegalovirus (CMV), an ubiquitous pathogen. Two CMV peptide pools (IE1 and pp65) were used to stimulate PBMCs and four effector functions were measured by multicolor flow cytometry (IL-2, TNFα, IFNγ and CD107a) to identify polyfunctional T cells. 17 healthy individuals, 13 patients with pre-dialysis chronic kidney disease (CKD) and 37 ESRD patients were enrolled in this study. All the donors were seropositive with an average age of 60. Patients with renal disease, especially the ESRD patients, showed increased levels of T cell differentiation, including the decrease in CD4+ and CD8+ TN cells and the increase in the CD4+ and CD8+ TEM and TEMRA cells. While the cellular frequency of CMV-reactive CD4+ and CD8+ T cells were similar among different groups, polyfunctional cell were dramatically reduced in ESRD. Among healthy individuals, CKD and ESRD patients, CD4+ CMV-IE1-reactive polyfunctional cell frequencies were 2.0%, 2.2%, and 0%, respectively (p=0.002). The CD8+ CMV-pp65-reactive polyfunctional cell frequencies were 12.4%, 8.8% and 0.8% in the three groups (p&lt;0.001). Such phenomenon resembles T cell exhaustion observed in HIV infection and further studies should investigate the level of inhibitory receptor expression levels such as PD-1 and CTLA-4 in these patients.
Characterization of keratinocytes isolated from human suction blisters and passaged with Rho kinase inhibitor
Authors: Datta Sandip K , Anderson Erik , Sastalla Inka , Moore Ian N. , Myles Ian Antheni 
Published: January 2023

Abstract
Keratinocytes are the most abundant cell type in the epidermis. They prevent desiccation and provide immunological and barrier mechanisms of defense against potential pathogens such as Staphylococcus aureus and Candida albicans. Study of this important first line of immune defense is hindered by the invasive isolation methods and insufficient techniques for long-term passage of primary keratinocytes in vitro. One method for obtaining primary human keratinocytes is to isolate them from blister roofs induced by negative pressure suction that separates the epidermis from the dermis in vivo in human subjects. This method allows collection of pure epidermal cells without dermal contamination in a minimally invasive manner. However, the method falls short because the isolated keratinocytes differentiate and senesce when cultured in vitro, with the maximum passage number being reported at 5. To address this problem of in vitro cultivation, we present evidence that the Rho kinase (ROCK) inhibitor Y-27632 can be used to effectively increase the proliferative capabilities of keratinocytes isolated using the suction-blister method, similar to what has been previously reported for neonatal foreskin-derived keratinocytes. We also show that upon removal of the ROCK inhibitor the cells retain their ability to stratify into an epithelium as well as produce human beta-defensins upon exposure to IL-17 and interferon-gamma. We expect that when coupled with the minimally invasive and efficient nature of the suction blister technique, our methodology could become a preferred procedure for keratinocyte isolation and cultivation that will allow analysis of keratinocyte function in both healthy volunteers and patients with diseases of interest.