Repetitive sulfur dioxide exposure in mice models post-deployment respiratory syndrome.

Soldiers deployed to Iraq and Afghanistan have a higher prevalence of respiratory symptoms than nondeployed military personnel and some have been shown to have a constellation of findings on lung biopsy termed post-deployment respiratory syndrome (PDRS). Since many of the subjects in this cohort reported exposure to sulfur dioxide (SO2), we developed a model of repetitive exposure to SO2 in mice that phenocopies many aspects of PDRS, including adaptive immune activation, airway wall remodeling, and pulmonary vascular (PV) disease. Although abnormalities in small airways were not sufficient to alter lung mechanics, PV remodeling resulted in the development of pulmonary hypertension and reduced exercise tolerance in SO2-exposed mice. SO2 exposure led to increased formation of isolevuglandins (isoLGs) adducts and superoxide dismutase 2 (SOD2) acetylation in endothelial cells, which were attenuated by treatment with the isoLG scavenger 2-hydroxybenzylamine acetate (2-HOBA). In addition, 2-HOBA treatment or Siruin-3 overexpression in a transgenic mouse model prevented vascular remodeling following SO2 exposure. In summary, our results indicate that repetitive SO2 exposure recapitulates many aspects of PDRS and that oxidative stress appears to mediate PV remodeling in this model. Together, these findings provide new insights regarding the critical mechanisms underlying PDRS.NEW & NOTEWORTHY We developed a mice model of "post-deployment respiratory syndrome" (PDRS), a condition in Veterans with unexplained exertional dyspnea. Our model successfully recapitulates many of the pathological and physiological features of the syndrome, revealing involvement of the ROS-isoLGs-Sirt3-SOD2 pathway in pulmonary vasculature pathology. Our study provides additional knowledge about effects and long-term consequences of sulfur dioxide exposure on the respiratory system, serving as a valuable tool for future PDRS research.

Multiplatform Single-Cell Analysis Identifies Immune Cell Types Enhanced in Pulmonary Fibrosis.

Immune cells have been implicated in idiopathic pulmonary fibrosis (IPF), but the phenotypes and effector mechanisms of these cells remain incompletely characterized. We performed mass cytometry to quantify immune cell subsets in lungs of 12 patients with IPF and 15 organ donors without chronic lung disease and used existing single-cell RNA-sequencing data to investigate transcriptional profiles of immune cells overrepresented in IPF. Among myeloid cells, we found increased numbers of alveolar macrophages (AMØs) and dendritic cells (DCs) in IPF, as well as a subset of monocyte-derived DCs. In contrast, monocyte-like cells and interstitial macrophages were reduced in IPF. Transcriptomic profiling identified an enrichment for IFN-γ response pathways in AMØs and DCs from IPF, as well as antigen processing in DCs and phagocytosis in AMØs. Among T cells, we identified three subsets of memory T cells that were increased in IPF, including CD4+ and CD8+ resident memory T cells (TRM) and CD8+ effector memory cells. The response to the IFN-γ pathway was enriched in CD4 TRM and CD8 TRM cells in IPF, together with T cell activation and immune response-regulating signaling pathways. Increased AMØs, DCs, and memory T cells were present in IPF lungs compared with control subjects. In IPF, these cells possess an activation profile indicating increased IFN-γ signaling and upregulation of adaptive immunity in the lungs. Together, these studies highlight critical features of the immunopathogenesis of IPF.

Cereblon harnesses Myc-dependent bioenergetics and activity of CD8+ T lymphocytes.

Immunomodulatory drugs, such as thalidomide and related compounds, potentiate T-cell effector functions. Cereblon (CRBN), a substrate receptor of the DDB1-cullin-RING E3 ubiquitin ligase complex, is the only molecular target for this drug class, where drug-induced, ubiquitin-dependent degradation of known "neosubstrates," such as IKAROS, AIOLOS, and CK1α, accounts for their biological activity. Far less clear is whether these CRBN E3 ligase-modulating compounds disrupt the endogenous functions of CRBN. We report that CRBN functions in a feedback loop that harnesses antigen-specific CD8+ T-cell effector responses. Specifically, Crbn deficiency in murine CD8+ T cells augments their central metabolism manifested as elevated bioenergetics, with supraphysiological levels of polyamines, secondary to enhanced glucose and amino acid transport, and with increased expression of metabolic enzymes, including the polyamine biosynthetic enzyme ornithine decarboxylase. Treatment with CRBN-modulating compounds similarly augments central metabolism of human CD8+ T cells. Notably, the metabolic control of CD8+ T cells by modulating compounds or Crbn deficiency is linked to increased and sustained expression of the master metabolic regulator MYC. Finally, Crbn-deficient T cells have augmented antigen-specific cytolytic activity vs melanoma tumor cells, ex vivo and in vivo, and drive accelerated and highly aggressive graft-versus-host disease. Therefore, CRBN functions to harness the activation of CD8+ T cells, and this phenotype can be exploited by treatment with drugs.

HDAC11 deficiency disrupts oncogene-induced hematopoiesis in myeloproliferative neoplasms.

Protein acetylation is an important contributor to cancer initiation. Histone deacetylase 6 (HDAC6) controls JAK2 translation and protein stability and has been implicated in JAK2-driven diseases best exemplified by myeloproliferative neoplasms (MPNs). By using novel classes of highly selective HDAC inhibitors and genetically deficient mouse models, we discovered that HDAC11 rather than HDAC6 is necessary for the proliferation and survival of oncogenic JAK2-driven MPN cells and patient samples. Notably, HDAC11 is variably expressed in primitive stem cells and is expressed largely upon lineage commitment. Although Hdac11is dispensable for normal homeostatic hematopoietic stem and progenitor cell differentiation based on chimeric bone marrow reconstitution, Hdac11 deficiency significantly reduced the abnormal megakaryocyte population, improved splenic architecture, reduced fibrosis, and increased survival in the MPLW515L-MPN mouse model during primary and secondary transplantation. Therefore, inhibitors of HDAC11 are an attractive therapy for treating patients with MPN. Although JAK2 inhibitor therapy provides substantial clinical benefit in MPN patients, the identification of alternative therapeutic targets is needed to reverse MPN pathogenesis and control malignant hematopoiesis. This study establishes HDAC11 as a unique type of target molecule that has therapeutic potential in MPN.

Repetitive Sulfur Dioxide Exposure in Mice Models Post-Deployment Respiratory Syndrome.

Soldiers deployed to Iraq and Afghanistan have a higher prevalence of respiratory symptoms than non-deployed military personnel and some have been shown to have a constellation of findings on lung biopsy termed post-deployment respiratory syndrome (PDRS). Since many of the deployers in this cohort reported exposure to sulfur dioxide (SO 2 ), we developed a model of repetitive exposure to SO 2 in mice that phenocopies many aspects of PDRS, including adaptive immune activation, airway wall remodeling, and pulmonary vascular disease (PVD). Although abnormalities in small airways were not sufficient to alter lung mechanics, PVD was associated with the development of pulmonary hypertension and reduced exercise tolerance in SO 2 exposed mice. Further, we used pharmacologic and genetic approaches to demonstrate a critical role for oxidative stress and isolevuglandins in mediating PVD in this model. In summary, our results indicate that repetitive SO 2 exposure recapitulates many aspects of PDRS and that oxidative stress may mediate PVD in this model, which may be helpful for future mechanistic studies examining the relationship between inhaled irritants, PVD, and PDRS.

The role of chitosan in wool finishing.

The application of the biopolymer chitosan as an alternative to the conventional contaminating processes in textiles was studied. As chitosan is produced by biomass, it is biodegradable and bioadsorbable. These properties are increasingly important given the current environmental legislation. The main aim of chitosan treatment of oxidised wool fabrics is to improve felting properties and dyeing behaviour.

Allelic diversity and affinity variants of MICA are imbalanced in Spanish patients with Behçet's disease.

The aetiology of Behçet's disease (BD) is still unknown, but genetic and environmental factors are involved. HLA-B*51 is considered a susceptibility marker and some MICA alleles have also been associated. Cytotoxic T lymphocytes have been suggested as responsible for BD lesions by engaging MICA through NKG2D surface molecules. In the present study, HLA-B and MICA alleles were typed by polymerase chain reaction using sequence-specific primers, in 165 healthy Spanish controls and 42 BD patients. In the healthy group, MICA*008 (28.48%), MICA*004 (17.58%), MICA*002 (14.24%) and MICA*009 (9.39%) were the predominant alleles and the most common haplotype was MICA*004-B*44 (12.12%). MICA*001 (5.15%), MICA*004, MICA*011 (4.54%) and MICA*018 (5.15%) were more frequent, and MICA*010 (1.81%) and MICA*008 were less prevalent than in other Caucasoid populations. Similar results have been reported in North African individuals and this could support the hypothesis of a common ancestral origin of both populations. The frequencies of MICA*009 and MICA*019 were significantly increased in our BD patients in comparison with controls: 22.62% versus 9.39% and 10.71% versus 1.81% respectively. The increase of MICA*019 had not been described in other BD cohorts, and it corroborates the genetic heterogeneity at MICA locus in BD patients. High-affinity MICA alleles for NKG2D were more frequent in controls than in patients. Moreover, high-affinity alleles were not found in homozygous BD patients. These results argue against the hypothesis of an autoaggressive response in BD patients through MICA-NKG2D interactions.

Elevated serum interleukin (IL)-12p40 levels in common variable immunodeficiency disease and decreased peripheral blood dendritic cells: analysis of IL-12p40 and interferon-gamma gene.

Common variable immunodeficiency disease (CVID) is a heterogeneous syndrome characterized by low immunoglobulin serum levels and recurrent bacterial infections. Several studies suggest that CVID patients have a polarized immune response towards a T helper type 1 phenotype (TH1). However, the factors causing the TH1 polarization remain to be determined in this disease. In the present study, serum interleukin (IL)-12, interferon (IFN)-gamma levels and the IL-12p40 and IFN-gamma gene were studied in CVID patients. Furthermore, we evaluate dendritic cells (DCs) compartment, myeloid dendritic cells (mDCs) and plasmocytoid dendritic cells (pDCs), which help to differentiate naive T cells preferentially into TH1 and TH2, respectively. The serum IL-12p40 subunit levels were increased significantly in CVID patients compared to healthy controls. We examined whether these elevated serum IL-12p40 levels are associated with IFN-gamma or IL-12p40 gene polymorphisms, or with new mutations in the IL-12p40 promoter gene. In our hands, no new mutations were found and gene polymorphisms frequencies in CVID patients were similar to the control population. In conclusion, the elevated serum levels of IL-12p40 found in our CVID patients were not related to these genetic variations. The DC compartment analysis did not show an imbalance between pDCs and mDCs, but revealed the presence of low numbers and percentage of both DC populations in CVID.

Blunted gamma delta T-lymphocyte response in chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) is characterised by an excessive inflammatory response to inhaled particles, mostly tobacco smoking. Although inflammation is present in all smokers, only a percentage of them develop COPD. T-lymphocytes are important effector and regulatory cells that participate actively in the inflammatory response of COPD. They comprise the T-cell receptor (TCR)-alpha beta (CD4+ and CD8+) and TCR-gamma delta T-lymphocytes. The latter represent a small percentage of the total T-cell population, but play a key role in tissue repair and mucosal homeostasis. To investigate TCR-alpha beta (CD4+ and CD8+) and TCR-gamma delta T-lymphocytes in COPD, the present authors determined, by flow cytometry, the distribution of both subpopulations in peripheral blood and bronchoalveolar lavage (BAL) samples obtained from patients with COPD, smokers with normal lung function and never-smokers. The present study found that: 1) the distribution of CD4+ and CD8+ lymphocytes in blood and BAL was similar in all three groups; 2) compared with nonsmokers, gamma delta T-lymphocytes were significantly increased in smokers with preserved lung function; and 3) this response was blunted in patients with COPD. These results highlight a novel, potentially relevant, pathogenic mechanism in chronic obstructive pulmonary disease.

[Pediatric AIDS through vertical transmission]. / SIDA pediátrico por transmisión vertical.

We report here the first paediatric immunodeficiency acquired syndrome (AIDS) in Spain, acquired by vertical-transmission and a second infant case with the same transmission and classified as AIDS related complex. We also mention the incidence and characteristics, of the neonatal population born in the last eighteen months to HIV positive mothers in our geographic area. The AIDS patients, was a female drug addicted parents born to who died at 22 month. Her parents are HIV positive and have ARC. The second patient was a boy, born to a sexual HIV positive partner of a member of a high risk group who died pat 17 month. Review of the literature until September 1986, revealed 369 AIDS paediatric patients, the number is lower 265 (7 Spanish), if only vertical transmitted AIDS is considered. For this reason, we considered pertinent to provide some information about clinical manifestations and laboratory, radiologic and necropsy findings in our patients.

Small cytoplasmic antigens from Pseudomonas aeruginosa stimulate gammadelta T lymphocytes.

Gammadelta T lymphocytes respond to different bacterial antigens and transformed cells. The antigenic molecules responsible for this activity have been studied extensively in antigenic preparations from Mycobacterium. We describe here the in vitro effect of Pseudomonas aeruginosa on gammadelta T lymphocytes and the properties of the implicated compounds. We found a preferential gammadelta T-cell expansion when we used heat-treated P. aeruginosa preparations and a dose-dependent inhibition of proliferation of peripheral blood mononuclear cells (PBMC) when non-heat-treated antigens were studied. This expansion corresponded to a Vgamma9-positive subpopulation. In contrast to alphabeta T lymphocytes, the highest stimulatory activity was restricted to very small cytosolic compounds. This activity was protease resistant and phosphatase sensitive and always dependent on interleukin (IL)-2 or alphabeta T-cell activation. We concluded that the antigenic molecules from P. aeruginosa that activated gammadelta T lymphocytes were small, non-peptidic, phosphorylated compounds, similar to those previously described from Mycobacterium.

Resultados del Taller de Autoinmunidad. Córdoba 2005 / No disponible

En el Congreso de la Sociedad Española de Inmunología, celebrado del 10 al 13 de mayo de 2005 en Córdoba, se analizaron 17 sueros procedentes de los 36 Laboratorios participantes en el XIX Taller de Autoinmunidad. Este año el interés se focalizó en marcadores de enfermedades autoinmunes hepáticas, fundamentalmente en las especificidades: AMA, LKM1, LC1, Actina-F, SLA, MND/Sp100, poro nuclear (gp210) y ANCA. Las técnicas empleadas fueron inmunofluorescencia indirecta (IFI), ELISA, western-blot, dot-blot e inmunoprecipitación. Se constató que son necesarias técnicas complementarias a la IFI, sobre todo en sueros de bajo título, imágenes solapadas o paraantígenos, como SLA, no discernibles por IFI. La técnica de dotblot,en general muy sensible, se mostró algunas veces insuficiente,en particular en el caso de la Actina-F y en otras dio falsos positivos por lo que debe ser interpretada también junto a otras técnicas. El western-blot, el dot-blot y el ELISA fueron más sensibles que la IFI en el estudio de los AMA. El western-blot con microsomas purificados fue la técnica menos sensible para LKM1.Se discutió la importancia de los anticuerpos anti-MND/Sp100 y poro nuclear para el diagnóstico de la cirrosis biliar primaria en ausencia de AMA. También se comentaron algunas especificidades concomitantes, fuera del contexto clínico analizado.En el ejercicio se incluyó por primera vez un CD con imágenes de IFI poco frecuentes, destinado a suplir aquellos sueros de volumen insuficiente para una amplia distribución. Si se generaliza esta iniciativa, puede llegar a constituirse un interesante archivo para consultas y docencia (AU)

During the Congress of the Spanish Society of Immunology, held in Córdoba from the 10th to 13th may 2005, the XIX Workshop on Autoimmune Diseases took place, focusing this year on the study of markers of hepatic autoimmune diseases. Seventeen sera were analysed, coming from the 36 participant laboratories. The sera included specificities like AMA, LKM1, LC1, F-actin,SLA, MND/Sp100, nuclear pore (gp210) and ANCA. The techniques used were indirect immunofluorescence (IIF), ELISA, western-blot, dot-blot and immunoprecipitation. The results allowed to state that techniques complementary to IIF are necessary to be applied, particularly in low titered sera, in overlapping patterns and in sera recognizing antigens, like SLA, not visible by IIF. Western-blot, dot-blot and ELISA M2 showed a higher sensitivity thanIIF for detecting AMA. F-actin was also underdetected by IIF. The western-blot performed with purified microsomes was less sensitive than dot-blot or ELISA for LMK1 detection. Dot-blot was a suitable technique but showed some false negative results (particularly in F-actin detection) and also false positive reactions and must be interpreted together with other techniques. Finally the discussion focused on the importance of anti-MND/Sp100 and nuclear pore complex for the diagnosis of primary biliary cirrhosis,particularly in the absence of AMA. Also some patterns,other than specific hepatic disease markers, were commented.To complete the exercise, for the first time, a CD was included containing some rare specificities of sera not available to bedistributed. If this initiative becomes established, an interesting archive for consultation and teaching may be created (AU)

Resultados del Taller de Autoinmunidad. Córdoba 2005 / Results from the XIX spanish autoimmunity workshop (Córdoba 2005)

En el Congreso de la Sociedad Española de Inmunología, celebradodel 10 al 13 de mayo de 2005 en Córdoba, se analizaron17 sueros procedentes de los 36 Laboratorios participantes en elXIX Taller de Autoinmunidad. Este año el interés se focalizó enmarcadores de enfermedades autoinmunes hepáticas, fundamentalmenteen las especificidades: AMA, LKM1, LC1, Actina-F, SLA, MND/Sp100, poro nuclear (gp210) y ANCA. Las técnicasempleadas fueron inmunofluorescencia indirecta (IFI), ELISA,western-blot, dot-blot e inmunoprecipitación.Se constató que son necesarias técnicas complementarias a laIFI, sobre todo en sueros de bajo título, imágenes solapadas o paraantígenos, como SLA, no discernibles por IFI. La técnica de dotblot,en general muy sensible, se mostró algunas veces insuficiente,en particular en el caso de la Actina-F y en otras dio falsospositivos por lo que debe ser interpretada también junto a otrastécnicas. El western-blot, el dot-blot y el ELISA fueron más sensiblesque la IFI en el estudio de los AMA. El western-blot conmicrosomas purificados fue la técnica menos sensible para LKM1.Se discutió la importancia de los anticuerpos anti-MND/Sp100y poro nuclear para el diagnóstico de la cirrosis biliar primaria enausencia de AMA. También se comentaron algunas especificidadesconcomitantes, fuera del contexto clínico analizado.En el ejercicio se incluyó por primera vez un CD con imágenesde IFI poco frecuentes, destinado a suplir aquellos suerosde volumen insuficiente para una amplia distribución. Si se generalizaesta iniciativa, puede llegar a constituirse un interesantearchivo para consultas y docencia

During the Congress of the Spanish Society of Immunology,held in Córdoba from the 10th to 13th may 2005, the XIX Workshopon Autoimmune Diseases took place, focusing this year onthe study of markers of hepatic autoimmune diseases. Seventeensera were analysed, coming from the 36 participant laboratories.The sera included specificities like AMA, LKM1, LC1, F-actin,SLA, MND/Sp100, nuclear pore (gp210) and ANCA. The techniquesused were indirect immunofluorescence (IIF), ELISA, western-blot, dot-blot and immunoprecipitation. The results allowedto state that techniques complementary to IIF are necessary to beapplied, particularly in low titered sera, in overlapping patternsand in sera recognizing antigens, like SLA, not visible by IIF. Western-blot, dot-blot and ELISA M2 showed a higher sensitivity thanIIF for detecting AMA. F-actin was also underdetected by IIF. Thewestern-blot performed with purified microsomes was less sensitivethan dot-blot or ELISA for LMK1 detection. Dot-blot was asuitable technique but showed some false negative results (particularlyin F-actin detection) and also false positive reactions andmust be interpreted together with other techniques. Finally thediscussion focused on the importance of anti-MND/Sp100 andnuclear pore complex for the diagnosis of primary biliary cirrhosis,particularly in the absence of AMA. Also some patterns,other than specific hepatic disease markers, were commented.To complete the exercise, for the first time, a CD was includedcontaining some rare specificities of sera not available to bedistributed. If this initiative becomes established, an interestingarchive for consultation and teaching may be created