Project description:To determine the different gene signatures between B lymphocytes from tumor draining lymph node (DLN) and normal lymph node (NLN), we have employed gene microarray as a discovery platform to identify gene signatures of tumor-educated B cells in DLN from tumor-bearing mice, taking NLN from normal mice as a control. We subcutaneously inoculated Balb/c mice with breast cancer cell line 4T1. Two weeks later, DLN was harvest and B cells were purified as descript in “treatment protocol”. From gene microarray, we found that B cells in DLN showed quite different transcript profiles from that in NLN.
Project description:We report the transcriptional profiles of human lung and lung draining lymph node T cell subsets that were sorted from paired samples. Naïve, central memory, effector memory and tissue resident memory CD4 and CD8 T cell subsets were sorted by FACS. Samples then underwent RNA sequencing to compare the similarities and differences between these T cell subsets and across tissues.
Project description:ISCOM vaccines induce a balanced Th1/Th2 response and cytotoxic T lymphocytes. The adjuvant component, ISCOM-Matrix, consists of purified saponin fractions, cholesterol and phospholipids. The mode of action for the ISCOM-Matrix is known to some extent but still we lack knowledge of important segments in initiation of the immune response. The study was performed to analyze the early transcriptional responses to the ISCOM-Matrix alone, without the use of co-administered antigen. Matrix M (AbISCO® 100) was given as intramuscular injection and after 24 hours the pigs was sacrificed for gene expression analysis, performed for the injection site and the draining lymph node. Results revealed 594 and 362 genes to be differentially expressed (FC > 2; q < 0.05) at the injection site and in the lymph node, respectively. This show that the ISCOM-Matrix, without any antigen added, has a clear immunomodulatory effect at the site of administration as well as in the draining lymph node. Six pigs, castrated males, 11 weeks old was used for the experiment. They were sorted two weeks prior the treatment and kept separate throughout the experiment. 3 pigs were intramuscularly (biceps femoris) injected with ISCOM-Matrix (AbISCO® 100) suspended in 1 ml sterile saline. 3 control animals were similarly injected with saline only. The pigs were sacrificed after 24 hours and muscle tissue from injection site and draining (internal iliac) lymph nodes were collected and transferred to RNAlater for RNA isolation.
Project description:We performed V(D)J scRNA sequencing to find transcriptional and B cell receptor repertoire differences in the germinal center of the draining lymph node of mice born to mothers chronically infected with Schistosoma mansoni. Cite-seq was used to help identify antigen specific cells.
Project description:ISCOM vaccines induce a balanced Th1/Th2 response and cytotoxic T lymphocytes. The adjuvant component, ISCOM-Matrix, consists of purified saponin fractions, cholesterol and phospholipids. The mode of action for the ISCOM-Matrix is known to some extent but still we lack knowledge of important segments in initiation of the immune response. The study was performed to analyze the early transcriptional responses to the ISCOM-Matrix alone, without the use of co-administered antigen. Matrix M (AbISCO® 100) was given as intramuscular injection and after 24 hours the pigs was sacrificed for gene expression analysis, performed for the injection site and the draining lymph node. Results revealed 594 and 362 genes to be differentially expressed (FC > 2; q < 0.05) at the injection site and in the lymph node, respectively. This show that the ISCOM-Matrix, without any antigen added, has a clear immunomodulatory effect at the site of administration as well as in the draining lymph node.
Project description:Germinal center (GC) like B cells can be generated in unstructured non-ectopic T and B cell infiltrates in the in inflamed lung. We used single cell RNA sequencing (scRNA-seq) for gene expression and B cell receptor (BCR) to compare antigen-specific B cells from lung to those from the lung-draining lymph node.
Project description:Lymph node (LN) stromal cells, particularly fibroblastic reticular cells (FRCs), provide critical structural support and regulate immunity, tolerance and transport properties of LNs. In many tumors, LN metastasis is predictive of poor prognosis, however, stromal contribution to the evolving microenvironment of tumor draining LNs (TDLN) remains poorly understood. Here we present comparative transcriptional data of resting and TDLN FRCs after different time points of tumor drainage. FRCs were isolated from lymph nodes and FACS sorted based on the expression of CD45-, CD31- and PDPN+
Project description:RNAseq was performed on human memory B cells isolated from lung, lung-draining lymph nodes and PBMCs to identify differentially expressed genes underpinning tissue localisation.