Project description:Determination of expression levels of light chain V genes in peripheral blood B cells after FACS sorting for two populations of B cells (CD20+CD138-IgKappa+IgLambda- and CD20+CD138-IgKappa-IgLambda+). Analysis was performed on healthy individuals and SLE patients with analysis performed using several models.
Project description:Determination of expression levels of light chain V genes in peripheral blood B cells after FACS sorting for two populations of B cells (CD20+CD138-IgKappa+IgLambda- and CD20+CD138-IgKappa-IgLambda+). Analysis was performed on healthy individuals and SLE patients with analysis performed using several models. Dual channel hybridization with experimental samples detected on red channel and reference sample detected on green channel. Two replicate hybridizations.
Project description:SLE patients are always with various disease manifestation. Various cytokines are pointed interacting and playing pathological roles in SLE although the etiopathology is still obscure. In this study, we aimed to investigate the effects of cytokine interactions in the immune response of SLE patients. Overexpressed interferon-inducible(IFI) genes were confirmed in peripheral blood from SLE patients. Using network-based analysis on the immune response-related genes, several networks including cytokines such as TNF and IFN-γ, or beta-estradiol(E2), were constructed. TNF-regulated genes were dominant in these networks but in vitro TNF stimulation on PBMCs showed no different responses in the expressions of these genes between SLE and healthy individuals. Co-stimulating experiments by TNF, IFN-γ, and E2 with IFN-α, revealed that TNF has repressive while IFN-γ essentially has synergistic effect with IFN-α on IFI gene expressions in vitro. E2 showed different effects on IFI gene expressions among 3 individuals. Peripheral blood was obtained from patients with SLE (n=11) and healthy women (n=6). Gene expression profile was analyzed using DNA microarray covering 30,000 human genes. Differentially expressed immune response-related genes were selected and analyzed by using Expression Analysis Systemic Explorer (EASE) based on Gene Ontology (GO) followed by network pathway analysis with Ingenuity Pathways Analysis (IPA).
Project description:B cells orchestrate the autoimmune responses in patients with systemic lupus erythematosus (SLE), but broad based B-cell directed therapies only show modest efficacy while attenuating humoral immune responses to vaccines and inducing acquired immunodeficiency. used proteomic and transcriptomic analyses of B cells from patients with SLE and healthy individuals and demonstrated a dominant DDR in SLE B cells.
2022-08-26 | PXD031389 | Pride
Project description:Naive B-cell receptor heavy chain repertoire of celiac patients and healthy controls
Project description:The immune responses to SARS-CoV-2 infections are very complex and remain to be further characterized. In this study, we performed the single-cell B-cell receptor (BCR) sequencing (scBCR-seq) of the PBMC samples from the healthy controls and SARS-CoV-2 infected individuals with various clinical presentations, and subsequently analyzed the abundance, diversity and difference of BCR repertoire in different groups of COVID-19 patients and the healthy controls, respectively. We identified a number of unique heavy or light chain VDJ genes pairs and combinational preference in each group, such as IGKV3-7 enriched in the asymptomatic subjects, whereas IGHV3-23-IGHJ4, IGHV1-18–IGLV3-19, IGHV1-18–IGLV3-21, and IGHV1-1–IGLV3-25 enriched in the recovery patients from the severe diseases. These findings may advance our understanding of the B-cell mediated immune responses to SARS-CoV-2 infections and help develop novel vaccine candidates and therapeutic antibodies against COVID-19.
Project description:Patients with systemic lupus erythematosus (SLE) have a markedly increased risk to develop cardiovascular disease, and traditional cardiovascular risk factors fail to account for this increased risk. We used microarray to probe the platelet transcriptome in individuals with SLE and healthy controls, and the gene and protein expression of a subset of differentially expressed genes was further investigated and correlated to platelet activation status. Real-time PCR was used to confirm a type I interferon (IFN) gene signature in patients with SLE, and the IFN-regulated proteins PRKRA, IFITM1 and CD69 (p<0.0001) were found to be up-regulated in platelets from SLE patients as compared to healthy volunteers. Notably, patients with a history of vascular disease had increased expression of type I IFN-regulated proteins as well as more activated platelets as compared with patients without vascular disease. We suggest that interferogenic immune complexes stimulate production of IFNα which up-regulates the megakaryocytic type I IFN-regulated genes and proteins. This could affect platelet activation and contribute to development of vascular disease in SLE. In addition, platelets with type I IFN signature could be a novel marker for vascular disease in SLE. Platelets were purified from SLE patients (n=10) and age and sex-matched healthy volunteers (n=10). cDNA was generated for each individual. For the microarray analysis, cDNA from the 10 SLE patients were pooled, as well as the cDNA from the 10 healthy volunteers to represent a mean mRNA expression level from 10 individuals.
Project description:Whole blood samples from SLE patients were collected and PBMCs were isolated using Ficoll-PaqueTM PLUS reagent according to the manufacturer’s protocol. The fresh isolated PBMCs from SLE individuals were treated with DMSO or spermine for 3h, PBMCs from healthy control were used as control, then samples were harvested and lysated with Trizol reagend for RNA-sequencing analysis.
Project description:The data corresponds to the analysis of T cell receptor (TCR) repertoires of FACS-purified Tstem, Tpex and TEM cells of six individuals. The analysis of the TCR beta chain (TRB) demonstrated the differences between Tstem and Tpex repertoire properties. In total, 36 samples were analyzed using the Human TCR Profiling Kit (MiLaboratory LLC) for sequencing libraries preparation and Illumina NextSeq 550 sequencing (150+150bp) followed by the demultiplexing procedure using MIGEC software.