Differential expression analysis of miR from individuals with follicular trachoma versus healthy controls. Results of DESeq differential expression analysis of sequencing read count data for normal healthy controls (N) versus cases of follicular trachoma (TF) with current Chlamydia trachomatis (Ct) infection (five samples of each). MiR with an average read count <5 were excluded. MRCâ=âMean read count. (XLS 73Â kb

Anna Last (3404372)

Chrissy Roberts (3404369)

David Mabey (37055)

Eunice Cassama (589482)

Martin Holland (728656)

Matthew Burton (3404375)

Meno Nabicassa (589483)

Robin Bailey (62493)

Sarah Burr (728655)

Tamsyn Derrick (389507)

BMC Infectious Diseases

PubMed

Differential expression analysis of miR from A2497 and A2497P- infected HCjE cells. Results of DESeq differential expression analysis of sequencing read count data for uninfected HCjE cells versus HCjE cells infected with A2497 and A2497P- strains of Chlamydia trachomatis at 48hpi. MiR with an average read count <5 were excluded. Three biological replicates of each condition were sequenced and analyzed. MRC = Mean read count, −VE = mock-infected cells. (XLS 137 kb

FigShare

Additional file 3: of Inverse relationship between microRNA-155 and -184 expression with increasing conjunctival inflammation during ocular Chlamydia trachomatis infection

Focused list of published roles for miR that are differentially expressed in follicular trachoma that relate to inflammation or fibrosis. (PDF 59Â kb

Additional file 8: of Inverse relationship between microRNA-155 and -184 expression with increasing conjunctival inflammation during ocular Chlamydia trachomatis infection

Additional file 6: of Inverse relationship between microRNA-155 and -184 expression with increasing conjunctival inflammation during ocular Chlamydia trachomatis infection

Chlamydial infection of HEp-2 and HCjE cell lines with three ocular Ct strains. HEp-2 cell monolayers infected with Ct strains A2497 (A), A2497P- (B) and A/HAR-13 (C), and HCjE cell monolayers infected with Ct strains A2497 (D), A2497P- (E) and A/HAR-13 (F) at 48 hpi. Cell nuclei are stained with DAPI (blue) and Ct inclusions are stained with anti-C. trachomatis MOMP antibody (FITC (green)). Cells were infected with an MOI of 1 and viewed at 40X magnification. Representative photographs were taken from one of three replicates for each condition. (PDF 770Â kb

Additional file 1: of Inverse relationship between microRNA-155 and -184 expression with increasing conjunctival inflammation during ocular Chlamydia trachomatis infection

Relative abundance of miR in the conjunctiva of cases of follicular trachoma (TF) and controls (N). MiR with a mean read count in TF cases or controlsâ<â5 were excluded. MiR were ranked by mean read count across all five samples of each phenotype group. (PDF 678Â kb

Additional file 5: of Inverse relationship between microRNA-155 and -184 expression with increasing conjunctival inflammation during ocular Chlamydia trachomatis infection

BACKGROUND: Trachoma, a preventable blinding eye disease, is initiated by ocular infection with Chlamydia trachomatis (Ct). We previously showed that microRNAs (miR) -147b and miR-1285 were up-regulated in inflammatory trachomatous scarring. During the initial stage of disease, follicular trachoma with current Ct infection, the differential expression of miR has not yet been investigated. METHODS: Conjunctival samples were collected from 163 children aged 1-9 years old living in a trachoma-endemic region of Guinea Bissau, West Africa. Small RNA sequencing (RNAseq) was carried out on samples from five children with follicular trachoma and current Ct infection and five children with healthy conjunctivae and no Ct infection. Small RNAseq was also carried out on human epithelial cell lines infected with ocular Ct strains A2497 and isogenic plasmid-free A2497 in vitro. Results were validated by quantitative PCR (qPCR) in 163 clinical samples. RESULTS: Differential expression of RNAseq data identified 12 miR with changes in relative expression during follicular trachoma, of which 9 were confirmed as differentially expressed by qPCR (miR-155, miR-150, miR-142, miR-181b, miR-181a, miR-342, miR-132, miR-4728 and miR-184). MiR-155 and miR-184 expression had a direct relationship with the degree of clinical inflammation. MiR-155 was up-regulated (OR = 2.533 ((95 % CI = 1.291-4.971); P = 0.0069) and miR-184 was down-regulated (OR = 0.416 ((95 % CI = 0.300-0.578); P = 1.61*10(-7)) as the severity of clinical inflammation increased. Differential miR expression was not detected in HEp-2 or HCjE epithelial cells 48 h post infection with Ct in vitro. HCjE cells, a conjunctival epithelial cell line, had a markedly different miR background expression compared to HEp-2 cells. CONCLUSIONS: In follicular trachoma, expression of miR-155 and miR-184 is correlated with the severity of inflammation. This likely reflects host regulation of the immune response and a prolonged period of wound healing following the clearance of Ct. Prolonged healing may be associated with subsequent development of scarring trachoma

Derrick, Tamsyn

Last, Anna R

Burr, Sarah E

Roberts, Chrissy H

Nabicassa, Meno

Cassama, Eunice

Bailey, Robin L

Mabey, David CW

Burton, Matthew J

Holland, Martin J

LSHTM Research Online

Inverse relationship between microRNA-155 and -184 expression with increasing conjunctival inflammation during ocular Chlamydia trachomatis infection.

Differential expression analysis of miR from A2497 and A2497P- infected HEp-2 cells. Results of DESeq differential expression analysis of sequencing read count data for uninfected HEp-2 cells versus HEp-2 cells infected with A2497 and A2497P- strains of Chlamydia trachomatis at 48hpi. MiR with an average read count <5 were excluded. Three biological replicates of each condition were sequenced and analyzed. MRC = Mean read count, −VE = mock-infected cells. (XLS 141 kb

Additional file 2: of Inverse relationship between microRNA-155 and -184 expression with increasing conjunctival inflammation during ocular Chlamydia trachomatis infection

Tamsyn Derrick

Anna R. Last

Sarah E. Burr

Chrissy h. Roberts

Meno Nabicassa

Eunice Cassama

Robin L. Bailey

David C. W. Mabey

Matthew J. Burton

Martin J. Holland

Crossref

Inverse relationship between microRNA-155 and -184 expression with increasing conjunctival inflammation during ocular Chlamydia trachomatis infection

Multivariable regression model of the contribution of miR expression to clinical papillary hypertrophy score. Collapsed papillary hypertrophy score (P0, P1 or P2/3, as defined by the WHO 1981 FPC scoring system) was used as an ordinal outcome variable to define trachomatous inflammation in 163 clinical samples. Age, gender, Ct load and inverted ∆CT values (40-∆CT) of miR are included as independent variables. Model AIC (Akaike information criterion) is 227.3725. aOR = Odds ratio, bCI = confidence intervals, cCt load is defined as log-(e) omcB copies/swab. (PDF 59 kb

Additional file 7: of Inverse relationship between microRNA-155 and -184 expression with increasing conjunctival inflammation during ocular Chlamydia trachomatis infection

https://figshare.com/articles/Additional_file_5_of_Inverse_relationship_between_microRNA-155_and_-184_expression_with_increasing_conjunctival_inflammation_during_ocular_Chlamydia_trachomatis_infection/4320707