Gene expression profiles in drug-induced hypersensitivity reactions with cutaneous involvement
ABSTRACT: Hypersensitivity reactions to medications constitute a growing problem in the clinical practice. In order to study the molecular basis underlying the pathogenesis of non-immediate hypersensitivity reactions to drugs, we characterized the gene expression profiles of PBMCs isolated from patients during the acute phase and upon resolution of the clinical symptoms using a cDNA array technology. Eighty five genes were found to be differentially expressed during the acute phase of drug-induced delayed hypersensitivity reactions. Furthermore, ninety two genes with distinct expression patterns during the acute phase of severe and benign diseases were identified. Keywords: Comparison between disease and healty status Overall design: Expression profiles of 11835 genes were analyzed in peripheral blood mononuclear cells from 13 patients with different clinical entities . Two samples were analyze from each patient. The first blood sample was drawn during the acute phase of the disease. The second blood sample was obtained upon resolutin of the clinical symptoms. The ratio between gene expression levels in both samples was calculated for each patient.
INSTRUMENT(S): Atlas Plastic Human 12K Microarray (Cat.#7931-1)
Project description:Hypersensitivity reactions to medications constitute a growing problem in the clinical practice. In order to study the molecular basis underlying the pathogenesis of non-immediate hypersensitivity reactions to drugs, we characterized the gene expression profiles of PBMCs isolated from patients during the acute phase and upon resolution of the clinical symptoms using a cDNA array technology. Eighty five genes were found to be differentially expressed during the acute phase of drug-induced delayed hypersensitivity reactions. Furthermore, ninety two genes with distinct expression patterns during the acute phase of severe and benign diseases were identified. Keywords: Comparison between disease and healty status Expression profiles of 11835 genes were analyzed in peripheral blood mononuclear cells from 13 patients with different clinical entities . Two samples were analyze from each patient. The first blood sample was drawn during the acute phase of the disease. The second blood sample was obtained upon resolutin of the clinical symptoms. The ratio between gene expression levels in both samples was calculated for each patient.
Project description:Nevirapine is a non-nucleoside reverse transcriptase inhibitor, a class of antiretroviral drug, used for the treatment of HIV-1 infection. Despite its wide use, nevirapine treatment has been associated with a significant incidence of different kind of hypersensitivity reactions (HSRs). We used microarrays to find significant genes that can relate to Nevirapine-persuaded hypersensitivity reactions in ‘acute’ patients compared to ‘recovered’ and/or ‘tolerant’ patients. Overall design: We have selected 18 patients’ samples, 6 from each group (Acute, Recovered, Tolerant) to investigate Nevirapine induced hypersensitivity reactions using transcriptomic profiling based on microarray technology. Acute:Patient taking nevirapine develops adverse drug reaction (ADR). Recovered: 6 weeks after acute reaction-not longer taking nevirapine. Tolerant: Patient on neviraoine and tolerating treatment.
Project description:Idiosyncratic drug reactions (IDRs) cause significant morbidity and mortality. In an animal model of IDRs, 50-80% of Brown Norway rats exposed to D-penicillamine develop an autoimmune syndrome after several weeks of treatment. The symptoms of the IDR are similar to that observed in humans who take D-penicillamine. The mechanism of this reaction is unknown, and no effective biomarkers have been identified to predict susceptibility. We postulate that cell stress caused by drugs is required to initiate the response. We used a highthroughput approach to identify factors that might represent danger signals by profiling hepatic gene expression 6 h after dosing with D-penicillamine (150 mg/kg). Our results show that the drug-treated animals cluster into two distinct groups. One group exhibits substantial expression changes relative to control animals. The most significantly altered transcripts have a role in stress, energy metabolism, acute phase response, and inflammation. We used quantitative reverse transcriptase polymerase chain reaction to measure transcript levels in liver biopsies of 33 rats and found that resistant animals cluster together. This 'resistant' cluster of animals contains 87.5% (7/8) resistant animals but only 48% (12/25) 'sensitive' animals. This separation is statistically significant at the p 0.01 level. Keywords: Response to Drug Overall design: Single-channel Affy arrays used to profile 4 control Brown Norway rats and 8 D-Penicillamine treated Brown Norway rats.
Project description:We used cDNA microarrays to systematically analyze host responses in SARS-infected individuals from onset of symptoms to discharge from hospital or death. We modeled gene expression in 60 datasets from 40 unique patients of varied clinical evolution with emphasis on correlating innate and adaptive immune responses with discrete clinical phases of the disease. Keywords: Comparative patient class comparision, SARS patients relative to healthy controls Overall design: Peripheral blood was collected in 5 ml heparinized blood collection tubes from study participants upon presentation to the hospital. The median length between onset of symptoms consistent with SARS and presentation to the hospital was 2 days (range 0-7 days). Peripheral blood specimens were then collected longitudinally every five days during hospitalization and at discharge from the hospital. Whole blood RNA was stabilized and purified using Paxgene Blood Collection Tubes and Paxgene RNA kits (Qiagen, Mississauga, ON, Canada). Sufficient RNA yields were obtained after Paxgene Whole Blood RNA isolation for 70 samples from 40 unique SARS patients and 10 healthy controls (HC1-10) and amplified using MessageAmp kits (Ambion). Replicate measurements or dye swaps were not possible due to severe limitations in RNA quantity. All samples (Cy5) were hybridized against amplified reference RNA (Cy3) (Stratagene, La Jolla, CA). Longitudinal ANOVA were performed to select significantly altered genes in the pre-crisis (acute) phase of SARS (0-8 days since onset, n=22), crisis phase for non-severe SARS patients (8+ days since onset, n=15) and crisis phase for severe SARS patients (8+ days since onset, n=23).
Project description:Approximately 10% to 20% of patients optimally treated for early Lyme disease develop persistent symptoms of unknown pathophysiology termed posttreatment Lyme disease syndrome (PTLDS). The objective of this study was to investigate associations between PTLDS and immune mediator levels during acute illness and at several time points following treatment. Seventy-six participants with physician-documented erythema migrans and 26 healthy controls with no history of Lyme disease were enrolled. Sixty-four cytokines, chemokines, and inflammatory markers were measured at each visit for a total of 6 visits over 1 year. An operationalized definition of PTLDS incorporating symptoms and functional impact was applied at 6 months and 1 year following treatment completion, and clinical outcome groups were defined as the return-to-health, symptoms-only, and PTLDS groups. Significance analysis of microarrays identified 7 of the 64 immune mediators to be differentially regulated by group. Generalized logit regressions controlling for potential confounders identified posttreatment levels of the T-cell chemokine CCL19 to be independently associated with clinical outcome group. Receiver operating characteristic analysis identified a CCL19 cutoff of >111.67 pg/ml at 1 month following treatment completion to be 82% sensitive and 83% specific for later PTLDS. We speculate that persistently elevated CCL19 levels among participants with PTLDS may reflect ongoing, immune-driven reactions at sites distal to secondary lymphoid tissue. Our findings suggest the relevance of CCL19 both during acute infection and as an immunologic risk factor for PTLDS during the posttreatment phase. Identification of a potential biomarker predictor for PTLDS provides the opportunity to better understand its pathophysiology and to develop early interventions in the context of appropriate and specific clinical information. Overall design: A total of 65 immune and inflammatory mediators were profiled in serum samples derived from early Lyme disease patients and age- and sex-matched controls. These samples have been generated as part of a prospective cohort study that includes a well-defined cohort of patients with acute Lyme disease enrolled from a Lyme endemic area of the mid-Atlantic United States. Only patients with untreated, confirmed early Lyme disease manifesting an active EM skin lesion at the time of enrollment, as defined by CDC case criteria are eligible. Patients with a history of prior Lyme disease or the presence of confounding preexisting medical conditions associated with prolonged fatigue, pain or neurocognitive symptoms are excluded. Controls are nonhospitalized age- and sex-matched and have no prior history of Lyme disease or any exclusionary medical conditions including lack of inflammatory disorders.
Project description:An antiepileptic drug carbamazepine is well tolerated by the majority of patients, but can cause severe and potentialy fatal hypersensitivity reactions in a small number of individuals. The aim of this study was to identify genetic predictors of hypersensitivity reactions to carbamazepine. We undertook a genome-wide association study (GWAS) in 22 patients of European ancestry with carbamazepine-induced hypersensitivity syndrome (HSS) and compared our data with genotypes from a healthy population within the WTCCC. We performed imputation of classical HLA alleles according to a recently described method (Science. 2010 Dec 10;330(6010):1551-7. Epub 2010 Nov 4). GWAS statistical analysis was performed using logistic regression with an additive model of inheritance.
Project description:Hypersensitivity reactions are rare, but potentially severe adverse effects of sulfonamide antibiotics. Increased in vitro toxicity of lymphocytes, primarily CD8+ T cells, to sulfonamide drug metabolites as been proposed as a marker for sulfonamide hypersensitivity, but the mechanisms underlying this marker are unknown. Therefore, we used microarrays to compare RNA expression of CD8+ T cell-enriched peripheral blood mononuclear cells of human patients who have had a hypersensitivity (HS) reaction to sulfonamide antibiotics vs. patients who have been tolerant (TOL) to a course of sulfonamide antibiotics. Overall design: 40ml of heparinized blood was obtained from each patient, peripheral blood mononuclear cells were extracted and enriched for CD8+ T cells and RNA was extracted.
Project description:Diagnosis of acute respiratory viral infection is currently based on clinical symptoms and pathogen detection. Use of host peripheral blood gene expression data to classify individuals with viral respiratory infection represents a novel means of infection diagnosis. We used microarrays to capture peripheral blood gene expression at baseline and time of peak symptoms in healthy volunteers infected intranasally with influenza A H3N2, respiratory syncytial virus or rhinovirus. We determined groups of coexpressed genes that accurately classified symptomatic versus asymptomatic individuals. We experimentally inoculated healthy volunteers with intranasal influenza, respiratory syncytial virus or rhinovirus. Symptoms were documented and peripheral blood samples drawn into PAXgene tubes for RNA isolation.
Project description:In comparison with allogeneic hematopoietic stem cell transplantation using other sources, umbilical cord blood transplantation (UCBT) offers substantial clinical advantages including a lower incidence and severity of acute graft-versus-host disease (GVHD) despite the use of allogeneic UCB stem cells with more disparate HLA. However, detailed pathophysiology of acute GVHD developed after UCBT has not yet been clarified. Here, we examined the roles of inflammatory cells in the pathogenesis of acute GVHD following UCBT, by expression profiling of a total of 615 genes in each of 4 subsets (CD4+, CD8+, CD14+, and CD56+) of peripheral blood mononuclear cells (PBMCs), which were taken from 5 patients with hematologic malignancy, in whom acute GVHD had developed after UCBT. By comparing expression profiles measured during acute GVHD with those measured upon discharge (recovery phase), for each subset of PBMCs, we identified immuno-regulatory genes which were assumed to be linked to the pathogenesis of acute GVHD. Keywords: disease state analysis Overall design: CD4+, CD8+, CD14+, and CD56+ cell subsets were isolated from the peripheral blood mononuclear cells (PBMCs) of 5 patients with hematologic malignancy, in whom acute GVHD had developed after cord blood transplantation. PBMCs were taken twice – during acute GVHD phase and upon discharge (recovery phase), from each patient. By using a custom-made fibrous oligonucleotide DNA microarray Genopal (Mitsubishi Rayon), we compared expression profiles of 615 unique genes measured during acute GVHD with those measured upon discharge (recovery phase), for each subset of PBMCs.