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ScRNAseq of the TCRs and BCRs of aqueous and blood immune cells in human uveitis

ABSTRACT: scRNAseq of the TCRs and BCRs of aqueous and blood immune cells in human uveitis

PROVIDER: PRJNA1146473 | ENA |

REPOSITORIES: ENA

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scRNAseq of the TCRs and BCRs of aqueous and blood immune cells in human uveitis

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Project description:Equine recurrent uveitis (ERU) is the only spontaneous model for recurrent autoimmune uveitis in humans, where T cells target retinal proteins. Differences between normal and autoaggressive lymphocytes were identified in this study by analyzing peripheral blood derived lymphocytes (PBL) proteomes from the same case with IRBP-induced uveitis sampled before (day 0), during (day 15) and after uveitic attack (day 23). Relative protein abundances of PBL were investigated in a quantitative, label-free differential proteome analysis in cells that were kept frozen for 14 years since the initial experiment. Quantitative data could be acquired for 2632 proteins at all three time-points. Profound changes (> 2 fold change) in PBL protein abundance were observed when comparing day 0 to 15, representing acute inflammation (234 regulated proteins) and day 0 to 23 (cessation; 382 regulated). Significant differences applied to proteins with functions in integrin signaling during active uveitis, involving “Erk and pi-3 kinase are necessary for collagen binding in corneal epithelia”, “Integrins in angiogenesis” and “Integrin-linked kinase signaling” pathways. In contrast, at cessation of uveitic attack, significantly changed proteins belonged to pathways of “nongenotropic androgen signaling”, “classical complement pathway” and “Amb2 integrin signaling”. Several members of respective pathways were earlier shown to be changed in naturally occurring uveitis, underscoring the significance of these findings here and proofing the value of the induced model in mimicking spontaneous autoimmune uveitis.

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Intra-ocular dendritic cells characterize HLA-B27-associated acute anterior uveitis

Project description:Uveitis describes a heterogeneous group of inflammatory eye diseases characterized by infiltration of leukocytes into the uveal tissues. uveitis associated with the HLA haplotype B27 (HLA-B27) is a common subtype of uveitis and a prototypical ocular immune-mediated disease. Local immune mechanisms driving human uveitis are poorly characterized mainly due to the limited available biomaterial and subsequent technical limitations. Here, we provide the first high-resolution characterization of intraocular leukocytes in HLA-B27 positive (n=3) and negative (n=2) anterior uveitis and an infectious endophthalmitis control (n=1) by combining single cell RNA-sequencing with flow cytometry and protein analysis. Ocular cell infiltrates consisted primarily of lymphocytes in both subtypes of uveitis and of myeloid cells in infectious endophthalmitis. HLA-B27 positive uveitis exclusively featured a plasmacytoid and classical dendritic cells (cDC) infiltrate. Moreover, cDCs were central in predicted local cell-cell communication. This suggests a unique pattern of ocular leukocyte infiltration in HLA-B27 positive uveitis with relevance of dendritic cells.

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