ABSTRACT: Purpose: Analysis at the single cell level of the CD4+ and CD8+ Lymphocytes T extracted and PBMCs from healthy human donor Methods: Single Cell RNAseq performed using the 10X genomics platform
Project description:PBMCs were extracted from 8 donors (an influenza, 2 mild and 2 severe COVID-19 patients, 3 healthy donors). Datasets generated by single-cell ATAC sequencing platform from 10X genomics. The library was generated by single-cell ATAC kit v1.1 following manufacturer's instructions.
Project description:The aim of the experiment was to investigate the clonal relationship between CD8+ T cell subsets of donor matched blood with epidermal CD8+ T cells. Abdominal skin (n=8) was separated into dermis and epidermis, followed by epidermal skin and PBMC isolation. Single CD8+ T cells (n=6) and CD3+ T cells (n=2) were FACS sorted, processed with the 5’ 10x Genomics workflow and sequenced at NGI Sweden.
Project description:The aim of the experiment was to understand the clonal relationship between CD8+ central memory blood T cells (TCM) and the in-vivo matured TCM tissue-resident memory-like T cells (TRM) on day 14. TCM were FACS sorted from human PBMCs (n=5). Half of the cells were processed on day 1 and half of the cells of each donor were matured into TRM with anti-CD3, IL-15 and TGF beta for 14 days. Both day 1 TCM and matured day 14 TRM-like cells were further processed using the 5´10X genomics workflow.
Project description:In this study, we investigated somatic mutations in T cells in patients with various hematological disorders. To analyze immune cell phenotypes with somatic mutations, we performed scRNA+TCRab sequencing from 9 patients with chronic GVHD and clonal expansions of CD4+ or CD8+ T cells based on T cell receptor sequencing. CD45+ PBMCs (lymphocytes and monocytes) were sorted with BD Influx cell sorter and subjected to sequencing with Chromium VDJ and Gene Expression platform (v1.1, 10X Genomics). Sequencing was performed with Novaseq 6000 (Illumina). The immune cell phenotypes were compared to healthy controls processed in the same laboratory (accession number E-MTAB-11170). Due to data privacy concerns, the raw sequencing data is in the European Genome-Phenome Archive (EGA) under accession code [xxxx] and can be requested through the EGA Data Access Committee.
Project description:In the clinical trial, we evaluated the immunological efficacy of the vaccination of GBM6-AD, a tumor-cell lysate, assisted with poly-ICLC in low grade glioma. To characterize the gene expression, subset proportions, and T-cell receptor (TCR) profile of T-cells in PBMCs, we analyzed pre- and post-neoadjuvant vaccinated PBMCs from four immunological responders (103-018, -26, -29, -51), using droplet-based 5’ single-cell RNA-sequencing (scRNA-seq) and single-cell T-cell receptor-sequencing (scTCR-seq) with 10x GENOMICS platform. Further, we analyzed the gene expression profiles in resected tumor specimens by bulk RNA-seq analyses.
Project description:5000 target cells per sample where sequenced from 2 gastric tumor samples and 1 sample of adjacent gastric mucosa Twist2Cre;Lkb1fl/+;RosaRTdtomato+/- mice. Drop-seq 10X Genomics (10X v3 library kit) was used for single cell barcoding and library preparation. Samples were sequenced with Novaseq6000.
Project description:Single-cell RNAseq (10x Genomics) analysis of human CD4+ T cells in IPEX patients, healthy donors and heterozygous mothers (blood). Human CD4+T cells from IPEX, HD and mothers were isolated from frozen peripheral blood mononuclear cells by flow cytometry as DAPI–CD3+CD4+ cells. In cohort 1, cells from separate donor were encapsulated in separate channel following 10x Genomics Single Cell 3′ Reagent Kit (V2 chemistry). In cohort 2, samples were tagged using a different Hashtags, pooled and encapsulared following 10x Genomics Single Cell 3′ Reagent Kit (V3 chemistry).
Project description:H2030-BrM cell line (lung adenocarcinoma, in vivo selected to increase its brain tropism) was injected intracardilly in three mice and when metastases were established as assesed by bioluminescence brains were microdissected to obtain GFP+ metastases. Metastases from the three mice were pooled and cell sorted for GFP. A pure GFP+ population was loaded onto a 10x Chromium Single Cell controller chip B (10x Genomics)