bluuebunny/crossref_metadata_2025_split huggingface dataset:
Intradermal RNA-Vaccination of Patients with Metastatic Renal Cell Carcinoma.
Authors: Wierecky Jan , Müller Martin R. , Häntschel Maik , Horger Marius S. , Pascolo Steve , Brugger Wolfram , Kanz Lothar , Brossart Peter 
Published: April 2020

Abstract
Background: The use of in vitro transcribed RNA isolated from tumor cells or coding for defined tumor associated antigens was shown to be a very powerfull method to generate antigen specific T cells upon transfection into dendritic cells (DC). More recently it was demonstrated in a mouse model that application of RNA intra dermaly can elicit both, CD8 and CD4 mediated immune responses. Furthermore, in Her-2-neu transgenic mice vaccinations with naked RNA were able to elicit a specific immune response in vivo and delay the development of breast cancer in treated animals. Based on these results we analyzed the clinical and immunological responses in patients with metastatic renal cell carcinoma (RCC) vaccinated with mRNA encoding for tumor associated antigens (TAA) using two different treatment arms.
Methods: In vitro transcribed RNA was generated using plasmids coding for the tumor antigens MUC1, CEA, Her-2/neu, telomerase, survivin and MAGE-1. RNA coding for HbsAg and Influenza matrix protein were included as controls. Vaccinations were performed intradermally on day 1,14, 28 and 42 in a first group of patients and on day 0–3, 7–10, 28 and 42 in a second group (intensified arm). Vaccinations were repeated afterwards monthly until tumor progression. One day after each RNA injection patients additionally received 1 injection of GM-CSF (250 μg) sc. The enhancement of T cell precursor was monitored using IFN-g ELISPOT and tetramer staining..
Results: 25 patients were included in this study. In 3 patients regression of metastases was induced and 6 patients had a stabilization of the disease. Specific CD8 and CD4 T cell responses in vivo were detected in the first analyzed patients. The treatment was well tolerated with no severe side effects. In most cases erythema and induration were observed after injections of GM-CSF. One patient developed an allergic exanthema after GM-CSF application.
Conclusions: This study demonstrates that intradermal RNA-vaccination can be effective in the treatment of metastatic RCC and induces clinical and immunological responses.
Clinical Evaluation of CCL18/PARC and Chitotriosidase Biomarkers in Gaucher Disease.
Authors: Alfonso Pilar , Latre Paz , de Blas Ignacio , Giraldo Pilar , Giralt Manuel , Pocovi Miguel 
Published: April 2020

Abstract
Gaucher’s disease (GD) is an autosomal recessive lysosomal storage disorder, characterized by accumulation of glycosphingolipid in so-called Gaucher cells. The clinical manifestations of Gaucher disease are highly variable, and although certain genotypes are often associated with mild or severe symtomps, a defined correlation between genotype and phenotype does not exist. Identification of serum biochemical markers characteristic of disease may be useful in the diagnosis and monitoring of GD, nevertheless about 6% of the population does not express the chitotriosidase (CT) gene. In this study, we analyzed using currently available enzyme analysis the relationship among two known surrogate markers: CT, which utility in initiation and optimization of costly therapeutic interventions has been highly demonstrated, and a newly-described chemokine, pulmonary and activation-regulated chemokine (CCL18/PARC) as associated to Gaucher disease.
Patients and methods: We have analysed 21 control samples, 149 samples of GD patients on enzyme replacement therapy (ERT), and 52 samples of GD patients on substrate reduction therapy (SRT), 4 samples of GD on combined therapy (ERT+SRT) and 7 samples of GD patients without therapy. The samples were stored at −80°C in the biobank of Biochemistry and Molecular and Cellular Biology Department of Zaragoza University. The CT activity and CCL18/PARC quantification were performed simultaneously in the samples obtained at baseline and yearly during 7 years under ERT.
Results: our results concluded that both markers levels similarly fell with time and their variations correlated strongly each other, mainly in patients under ERT. The poor correlation of both variables in the case of SRT might be due to small number of samples. These findings demonstrated that CCL18 is a good biomarker of monitoring GD, comparable to CT and very useful in patients without expression of CT gene.
Conclusion: The availability of sensitive plasma surrogate markers may be of great value for monitoring the efficacy of treatment, especially in cases of deficiencies of some marker.
Gene Expression Profiling of Adenosine Signaling in the Human Monocytic THP-1 Cells.
Authors: Chen Chin-Fu , Cheng Chun-Huai 
Published: April 2020

Abstract
Inflammation, a host response to infection or injury, is usually beneficial to the host. However, uncontrolled or prolonged inflammation can cause tissue and organ damage, and possibly lead to diseases like atherosclerosis, rheumatoid arthritis, asthma, and cancers. Much attention has focused on pro-inflammatory signaling but little is known about the mechanisms that resolve inflammation (anti-inflammation). Adenosine, released from leukocytes and endothelial cells, is an endogenous anti-inflammatory mediator that down-regulates acute inflammation in part by modulating macrophage cytokine production. LPS (lipopolysaccharide) is a potent pro-inflammatory reagent of macrophages that acts primarily through Toll-like receptors (TLRs). Studies using murine cells have showed that co-stimulation of both adenosine receptors and TLRs greatly increases the production of vascular endothelial growth factor (VEGF). The mechanisms for this synergistic interaction are currently not known. It is also unknown whether the adenosine-LPS interaction occurs in human macrophages. To gain insights and to generate new hypotheses, we have utilized oligonucleotide microarrays to examine how adenosine alters global gene expression in the human monocytic THP-1 cell line. We analyzed differentially expressed genes induced by adenosine alone, by LPS alone, and by adenosine in the presence of LPS. The genes activated by adenosine are enriched in processes transcription, RNA modification and cellular defense response, but are lacking processes involved in lipid biosynthesis and protein catabolism. Our result shows that at the mRNA level adenosine alone induces a nine-fold increase of VEGF, and adenosine in the presence of LPS elevates VEGF expression three fold when compared to controls. Many genes with the greatest induction by adenosine in the presence of LPS have unknown functions in macrophages. The genes with known functions include transcription regulation (EGR1, HEY1, and CTBP1), nuclear receptors (NR4A2), immune response modulators (IL1A, STAT4, and INHBA), apoptosis facilitator (BCL2L11), signal transducers (PRPF4B, LNK, and ARHGEF7), and glutamate transporter (SLC1A2). These results suggest that the mechanisms by which adenosine mediates its anti-inflammatory signals are far more complex than simply regulating cytokine production. In addition, we found that three TLR signaling genes are uniquely suppressed by adenosine in the presence of LPS: CREB1, TLR1, and IRF2. CREB1 modulates transcription of cAMP signaling; TLR1 is part of the TLR2/TLR1 complex and can regulate TLR2 mediated pathways; IRF2 is a mediator of interferon signaling. We are in the process of verifying these observations using real-time PCR and independent RNA samples.
Intravenous gamma Globulin Does Not Increase RNA Expression of FcgRIIb in Purified Human Monocytes.
Authors: Chan Howard H. , Su Brenda , Li Wei , Kelton John G. , Denomme Greg 
Published: April 2020

Abstract
Introduction
Intravenous gamma globulin (IVIG) has been shown to modulate immune cytopenia through FcγRIIb in a murine model (1). There are conflicting results from the studies on the protein expression of the FcγRIIb. In that murine model, there is increase in FcγRIIb positive splenocytes following IVIG treatment (1). Yet, in patients with Kawasaki disease, flow cytometric analysis of peripheral CD14+ mononuclear cells shows no change in FcγRIIb expression (2). To study the effect of IVIG on the RNA expression of FcγIIB, we performed RT-PCR using monocytes collected from three healthy volunteers. The monocytes were harvested using a magnetic separation method. The purified monocytes were then cultured with IVIG in vitro.
Methods
Peripheral blood mononuclear cells (PBMC) from three healthy donors were isolated from heparin anticoagulated blood by Ficoll-Hypaque density separation. The PBMC were washed and the monocytes were isolated using anti-CD14 magnetic bead positive selection. In the first set of experiments, purified monocytes were cultured at 37°C, 0.5% CO2 in media containing 10% fetal calf serum (FCS) (control) and in the same media containing IVIG (0.5 mg/mL and 5.0 mg/mL). In the second set of experiemnts, purified monocytes were cultured at 37°C, 0.5% CO2 in serum free medium (SFM), in SFM containing IVIG (5.0 mg/mL), and in SFM containing 10% FCS. Monocytes were harvested from the culture flasks after an 18-hour incubation. Isolated monocytes were lyzed and the total RNA was extracted with TRIzol reagent according to the manufacturer’s protocol. One μg of RNA was used to generate first strand complimentary DNA using oligo-dT and Superscript II reverse transcriptase. Amplitaq Gold DNA polymerase was used to PCR-amplify FCGR2B cDNA. PCR amplification was performed using 5 μL of cDNA and 1 μM each of an FCGR2B-specific primer pair (3) in a total volume of 50 μL. The PCR products were evaluated using 10% polyacylamide gel electrophoresis.
Results
There was no change in FCGR2B expression when purified monocytes were incubated with either 0.5 mg/mL or 5mg/mL IVIG for 18 hours (n=3) (Fig1).
However, there was a demonstrable increase in FCGR2B expression when monocytes were cultured in the presence of FCS (Fig2).
Conclusion
IVIG does not directly modulate monocyte FCGR2B gene transcription.
Figure Figure
Hemophagocytic Lymphohistiocytosis in a Extremely Low Birth Weight Infant Presenting with a Neonatal Thrombocytopenia.
Authors: Lee Mee Jeong , An Young Won , Chang Young Pyo , Lee Won Ae , Park Joowan , Lee Young Suk , Ghim Thad T. 
Published: April 2020

Abstract
INTRODUCTION: Hemophagocytic lymphohistiocytosis(HLH) is a potentially fatal disease presenting at an young children. Symptoms and physical findings include fever, rash and pancytopenia associated with histiocytic infiltration of bone marrow, liver and spleen. Its occurrence in neonates is very rare. We describe a female infant diagnosed with HLH, who was born at 24 week 3 days of gestational age(GA) weighing 666 gm. The infant expired due to aggravating thrombocytopenia and hepatosplenomegaly leading to multiple organ failure associated with suspected sepsis. Autopsy findings lead to a confirmation of HLH.
CASE: The baby, weighing 666gm, was born from a 36 year-old mother at GA 24 weeks 3 days, by emergency cesarian section due to vaginal bleeding. The mother came from the Republic of Kyrgystan and didn’t receive any antenatal care. The mother had a history of 8 prior abortions due to incompetence of internal os cervix (0–5–3/0–0). CBC on the first day was normal, but on the third day itg showed thrombocytopenia (Hb 11.2 g/dL, WBC 11,700/mm3, Platelet 78x103/mm3). Antibiotics were administered for suspected sepsis. On the 47th day, abdominal ultrasonography was performed due to abdominal distension and TPN-induced conjugated hyperbilirubinemia. GB wall edema and hepatomegaly were noted. Thrombocytopenia persisted and on the 55th day the patient became hypotensive and develped metabolic acidosis. Laboratory work up for persistent thrombocytopenia included anti-platelet antibody(−), TORCH(−), parvovirus-B19 PCR(-), CMV PCR (−), EBV infection(−). Bone marrow biopsy was not done. The patient expired from sepsis and multiorgan failure on the 61st day. Autopsy findings showed histiocyte infiltration of all tissue including the liver, spleen and brain. Bone marrow biopsy also showed infiltration of abundant hemophagocytic histiocytes consistent with a diagnosis of HLH.
CONCLUSION: A premature newborn with HLH described here presented with thrombocytopenia. Therefore, HLH should be included in the differential diagnosis of neonatal thrombocytopenia.
The Role of Endothelial PI3Kγ Activity in Neutrophil Trafficking.
Authors: Diacovo Thomas G. , Doggett Teresa A. , Huang Ching-Yu , Douangpanya Jason , Hayflick Joel , Turner Martin , Penninger Josef , Puri Kamal 
Published: April 2020

Abstract
Phosphoinositide 3-kinase gamma (PI3Kγ) in neutrophils plays a critical role in the directed-migration of these cells into inflamed tissues. In this study, we demonstrate the importance of the endothelial component of PI3Kγ activity relative to its leukocyte counterpart in supporting neutrophil interactions with the inflamed vessel wall. Despite the reconstitution of class Ib PI3K function in neutrophils of p110γ−/− mice, we observed a 45% reduction in accumulation of these cells in an acute lung injury model. Mechanistically, this appears to result from a perturbation in selectin-mediated adhesion as manifested by a 70% reduction in WT neutrophil attachment to and 17-fold increase in rolling velocities on p110γ−/− microvessels in vivo in response to TNFα. This alteration in adhesion was further augmented by a deficiency in p110δ, suggesting that the activity of both catalytic subunits is required for efficient capture of neutrophils by cytokine-stimulated endothelium. Interestingly, E-selectin-mediated adhesion in p110gamma]−/− mice was impaired by &gt; 95%, but no defect in NF-κB-induced gene expression was observed. These findings suggest a previously unrecognized partnership between class I PI3Ks expressed in leukocytes and endothelium, the combination of which is required for the efficient trafficking of immunocompetent cells to sites of inflammation.
Moribund Blood Monocytes: Apoptosis Limits the Yield of Monocyte Derived Dentritic Cells from Apheresis Samples.
Authors: Macke Lars , Geffers Robert , Rohde Manfred , Garritsen Henk S. , Woermann Bernhard , Lindenmaier Werner , Dittmar Kurt E. 
Published: April 2020

Abstract
Cells of the monocyte/macrophage/dendritic cell (DC) lineage play a central role in inflammation and form the bridge between innate and acquired immunity. In recent years monocytes also found special attention as source of precursor cells for the generation of monocyte derived dendritic cells for immunotherapy. However, monocytes circulate in the blood stream only temporarily with a half life of about 24 h and die spontaneously from apoptosis in the absence of specific survival signals. When we investigated the survival of blood monocytes isolated by CD14 positive selection from apheresis samples and cultivated in the presence of GM-CSF/IL4, we observed a loss of viability of about 66% of the cells within 48h. This would be consistent with the view that about 80% of the monocytes are bound to die at the time of isolation. Detailed flow cytometric analysis revealed that loss of vital cells is accompanied by the appearance of a CD14-negative population, which stained positive for annexin V. To analyse the fate of these cell populations CD14-positive and CD14-negative cells were separated by cell sorting at day 1 and day 2. Flow cytometry and electron microscopy identified predominantly dying cells in the CD14-negative population, whereas a large fraction of the CD14-positive population survived. The CD14-positive cells can be differentiated with an efficiency of 64%–91% to mature DC after cultivation under DC generating conditions; these include the presence of GM-CSF and IL4 and addition of a maturation mix (TNFa, IL1b, IL6 and PGE2) on day 5. DC generated from sorted CD14-positive cell populations were qualitatively and quantitatively comparable to DC from unsorted monocytes. To study the genes involved we analyzed the transcriptome on each cultivation day using Affymetrix HG-U133A DNA-Arrays. During the first 24h, the expressions of about 4700 genes were regulated. About 3000 genes were up regulated and about 1700 genes were down regulated. Especially, we focused on the expression of genes involved in the apoptosis pathway during the early phase of cultivation. Among apoptosis related genes caspase 3, a central player in natural apoptosis of monocytes, was up regulated on day 1. In summary these data suggest that apoptosis limits the yield of monocyte derived DC and that cultivation in the presence of caspase inhibitors or survival signals could improve the efficiency of generation of monocyte derived dendritic cells.
Deficiency of 2,3-Dimethoxy-5-Methyl-6-Decaprenyl-1,4-Benzoquinone (Mitoquinone) and Hyperlactic Acidosis in Human Immunodeficiency Virus (HIV) Patients with Head and Neck Squamous Cell Carcinoma (HNSCC) Treated with Nucleoside Reverse Transcriptase Inhibitor (NRTI)-Based HAART and Chemoradiation.
Authors: McPherson Eugene , Mejia E. , Hazel E. , Tassy P. 
Published: April 2020

Abstract
Mitoquinone is a nutrient obtained fromm food and produced in small amounts by the body with a crucial component of oxidative phosphorylation process in the mitochondria generating adenosine triphosphate (ATP). Deficiency of mitoquinone has been described based on failure of biosynthesis caused by gene mutation, inhibition of biosynthesis by HMG coA reductase(statins), compromisal of immune system function in breast cancer and acquired immunodeficiency syndrome (AIDS). Patients with (HNSCC) coinfected with HIV and deficient of mitoquinone, progress more rapidly to AIDS and when treated with statins for dyslipidemia, mitoquinone can become severely depressed and cause serious fatal type B lactic acidosis requiring emergent interventions. Correction of mitoquinone deficiency by supplementing mitoquinone may prevent and reverse the lactic acidosis seen in NRTI-based HAART (Highly Active Antiretroviral Therapy) allowing more effective chemoradiation with less side effects of mucositis, anemia and dermatitis. We present a case of a 44 year old female with HIV disease who developed a right tongue base squamous cell carcinoma (T2N0M0) lesion with trismus, elevated epstein-barr virus DNA (EBVDNA-PCR), dyslipidemia, and renal insufficiency was worked up and staged and treatment started with radiation therapy (XRT). No platinum chemotherapy given due to severe renal disease). Seven weeks of radiation therapy was given with interruption caused by grade II-III mucositis overcome by mitoquinone adjuvant therapy. Immune parameters CD4+ level initially was 71/cumm (10%) with a CD4/CD8 ratio 0f 0.17 and CD16+, CD56+ of 143/cumm (20%). A good response to XRT with less trismus and tumour shrinkage but soon after XRT she developed lactic acidosis with evidence of mitoquinone deficiency, 2.45 mmol/L and 0.4 mg/L respectively. Apolipoprotein A-1 level decreased along with high density lipoprotein-cholesterol (HDL-C). Statin therapy was started and lactic acidosis persisted with no further deteriorations of parameters. Lactic acidosis and mucositis (grade I) improved with mitoquinone supplements. The CD4+ level increased to 132/cumm (20%) and CD16+, CD56+ increased to 298/cumm (26%). CONCLUSIONS:HIV disease patients with HNSCC malignancy and mitoquinone deficiency may be at increase risk for mitochondrial DNA toxicity, chemoradiation side effects and severe lactic acidosis when treated with NRTI-based HAART, statins and XRT. Mitoquinone supplementation may prevent and reverse fatal lactic acidosis, modulate immune status and improve response to chemoradiation therapy, survival and quality of life.
Open-Label Study of Cellular Immunity in Dialysis Patients after Intravenous Ibandronate.
Authors: Bergner Raoul , Weiss Burkhard , Hoffman Martin , Henrich Dirk , Uppenkamp Michael 
Published: April 2020

AbstractIntravenous infusion of bisphosphonates can cause acute phase reactions which may be caused by changes in the cellular immune system, particularly in lymphocyte subtypes. These immune changes and an acute phase reaction both normally disappear after 48 hours. Previous studies have shown that kidney transplant recipients treated with immunosuppressive therapy and ibandronate for bone protection have a lower rejection rate than those treated with immunosuppressive therapy and placebo.1 This suggests that there may be long-term changes in the immune system occurring with ibandronate treatment. We investigated the long term changes in the cellular immune system during ibandronate treatment in dialysis patients. As these patients are not normally treated with immunosuppressive or cytotoxic agents, which can cause changes in the cellular immune system, they are an eligible population for such a study. In this open-label trial, 16 patients with end-stage renal disease receiving regular hemodialysis were recruited. All patients were treated with ibandronate 2mg every 4 weeks for renal osteopathy; a dose that provides similar AUC and bone exposure as a 6mg dose in patients with normal renal function. The cellular immune system was investigated before first ibandronate application and the measurement was repeated at weeks 2, 4 and 48. The following parameters were measured by blood count differentiation: leucocytes, granulocytes lymphocytes, monocytes. Lymphocyte subtypes were measured by flow cytometry: B-lymphocytes (CD3+/CD19+), T-helper cells (CD3+/CD4+), T-suppressor cells (CD3+/CD8+), natural killer (NK)-cells (CD3+/CD16+56+), helper-inducer cells (CD4+/CD29+), activated T-cells (CD3+/HLA-DR+), activated T-lymphocytes (CD3+/CD25+), naive T-cells (CD3+/CD45RA+) and memory-T-lymphocytes (CD3+/CD45RO+). Twelve patients completed the study and were evaluated. One patient dropped out because of flu-like symptoms with muscle pain after the first ibandronate infusion; however this was well controlled with paracetamol. Three patients died due to concomitant diseases (diabetes and cardiovascular events). There were no statistically significant differences in cellular immunity over time as measured in weeks 0, 2, 4 and 48 (see Figure). In this small-pilot study of dialysis patients receiving ibandronate, no changes in the cellular immune system were observed over time. Changes in different lymphocyte subtypes, which occur in the acute phase reaction after first infusion, were not seen. Reduced rejection rate in transplant recipients after ibandronate infusion cannot be explained by changes in the cellular immune system, and must therefore occur by another mechanism.Figure Figure
A Shared Gene-Expression Signature in Innate-Like Lymphocytes.
Authors: Yamagata Tetsuya , Benoist Christophe , Mathis Diane 
Published: April 2020

Abstract
Innate and adaptive immunity are the two major arms of the immune system. They rely on very distinct cell-types, primarily distinguished by the source of diversity for non-self recognition, of germline or somatic origin. There exists, however, a subset of lymphocytes whose receptors require rearrangement but result in semi-invariant structures with a high degree of self-specificity. We hypothesized that these innate-like lymphocytes might share a common gene transcription signature. To test this notion, we made pair-wise comparisons of the gene-expression profiles of innate-like lymphocytes and closely paired adaptive system counterparts (NKT vs. CD4T, CD8ααT vs. CD8αβT, B1 vs. B2), and bioinformatically extracted common features and common genes distinguishing innate from adaptive cell-types. A statistically significant “innate signature” was indeed distilled, composed of a small set of genes over- and under-expressed in innate vs. adaptive lymphocytes. Particularly intriguing was the high representation of interferon-inducible GTPases crucial for resistance against intracellular pathogens, and of small G proteins involved in intracellular vacuole maturation and trafficking. Overall, this combined expression pattern can thus be designated as an “innate signature” among lymphocytes.