INT340809

Context Info Confidence 0.77 First Reported 2010 Last Reported 2010 Negated 0 Speculated 1 Reported most in Body Documents 1 Total Number 41 Disease Relevance 27.14 Pain Relevance 1.69

Anatomy Link Frequency T cells 7 cornea 6 blood 5 stroma 2 corneal epithelium 2 Btla (Mus musculus)

Pain Link Frequency Relevance Heat Inflammation 615 99.76 imagery 41 98.04 cytokine 328 93.12 Inflammatory response 41 77.76 cINOD 41 7.04 corticosteroid 41 5.00

Disease Link Frequency Relevance Heat Infection 1599 99.92 INFLAMMATION 697 99.76 Cold Sores 2173 99.64 Keratitis 1517 99.26 Sprains And Strains 246 98.92 Herpes Simplex Virus 123 97.32 Disease 328 96.96 Eye Disease 82 96.24 Herpes Simplex Virus Infection 123 93.72 Necrosis 41 93.32

Sentences Mentioned In

Key:

Protein

Mutation

Event

Anatomy

Negation

Speculation

Pain term

Disease term

To confirm whether BTLA takes part in the immunopathogenesis of HSK in BALB/c mice, we first tested the expression of BTLA in the cornea and peripheral blood of HSV-1 infected mice.

Gene_expression (expression) of BTLA in blood associated with cold sores and keratitis

1) Confidence 0.77 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0.87 Pain Relevance 0.04

However, BTLA expression did not increase in CD8+ T cells after HSV-1 infection (Figure 1C).

Gene_expression (expression) of BTLA in T cells associated with cold sores and infection

2) Confidence 0.77 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 1.06 Pain Relevance 0.07

In the normal uninfected mouse cornea, BTLA expression was only detected in the corneal epithelium (Figure 1D).

Gene_expression (expression) of BTLA in corneal epithelium

3) Confidence 0.77 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 1.03 Pain Relevance 0.07

Immunohistochemistry results for BTLA expression in the corneas are shown in Figure 1D.

Gene_expression (expression) of BTLA in corneas

4) Confidence 0.77 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 1.05 Pain Relevance 0.07

Immunohistochemical detection of BTLA expression in murine corneas after HSV-1 infection

Gene_expression (expression) of BTLA in corneas associated with cold sores and infection

5) Confidence 0.77 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0.29 Pain Relevance 0

As shown in Figure 3A, at 48 h post-transfection, the immunofluorescence intensity was high in HEK293 cells transfected with pBTLA.

Gene_expression (transfected) of pBTLA

6) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0.49 Pain Relevance 0.04

BTLA expression is upregulated in the murine peripheral blood and cornea after HSV-1 infection

Gene_expression (expression) of BTLA in blood associated with cold sores and infection

7) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0.79 Pain Relevance 0.05

The restriction endonuclease digestion and sequence analyses confirmed successful construction of the recombinant expression plasmid pcDNA-BTLA (named pBTLA).

Gene_expression (expression) of pBTLA

8) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0.46 Pain Relevance 0.04

The results indicate that the target protein BTLA was expressed successfully.

Gene_expression (expressed) of BTLA

9) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0.65 Pain Relevance 0.09

The recombinant pBTLA was transfected into the HEK293 cell line (the Institute of Genetics of Fudan University, Shanghai, China) to express the target protein BTLA, which was detected by indirect immunofluorescence and western blotting.

Gene_expression (express) of BTLA

10) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0 Pain Relevance 0

A representative plot gated on CD4+ T cells demonstrated higher BTLA expression (20.14%) on day 10 after corneal inoculation with HSV-1 compared to day 0 (3.10%, Figure 1B).

Gene_expression (expression) of BTLA in T cells associated with cold sores

11) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 1.00 Pain Relevance 0.06

The restriction endonuclease digestion and sequence analyses confirmed successful construction of the recombinant expression plasmid pcDNA-BTLA (named pBTLA).

Gene_expression (expression) of pcDNA-BTLA

12) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0.46 Pain Relevance 0.04

BTLA expression increased in CD4+ T cells, peaking on day 10 after infection (Figure 1A).

Gene_expression (expression) of BTLA in T cells associated with infection

13) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0.93 Pain Relevance 0.04

However, in corneal tissues derived from HSV-1 infected mice, BTLA expression was increased in the corneal epithelium, stroma, and endothelium (Figure 1E).

Gene_expression (expression) of BTLA in corneal epithelium associated with cold sores

14) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 1.01 Pain Relevance 0.07

Forty-eight hours after transfection, the cells were fixed with 100% acetone at –20 °C for 15 min and BTLA expression was detected using the indirect immunofluorescence assay, with a monoclonal antibody against the BTLA protein and a FITC-labeled rabbit-anti-mouse IgG (Sigma, St.

Gene_expression (expression) of BTLA

15) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0 Pain Relevance 0

The recombinant pBTLA was transfected into the HEK293 cell line (the Institute of Genetics of Fudan University, Shanghai, China) to express the target protein BTLA, which was detected by indirect immunofluorescence and western blotting.

Gene_expression (transfected) of pBTLA

16) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0 Pain Relevance 0

Expression level of the recombinant plasmid pBTLA

Gene_expression (Expression) of pBTLA

17) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0.45 Pain Relevance 0.04

In addition, the stroma of infected cornea contained many inflammatory cells that expressed BTLA (Figure 1E).

Gene_expression (expressed) of BTLA in stroma associated with inflammation

18) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0.90 Pain Relevance 0.05

l connection system was obtained with the BTLA gene and pcDNA3.1 (BTLA:pcDNA3.1=1:1): pcDNA3.1 segment (4 ?

Gene_expression (gene) of BTLA

19) Confidence 0.67 Published 2010 Journal Molecular Vision Section Body Doc Link PMC2965573 Disease Relevance 0 Pain Relevance 0

BTLA expression in the infected cornea was detected by immunohistochemistry.

Gene_expression (expression) of BTLA in cornea

20) Confidence 0.60 Published 2010 Journal Molecular Vision Section Abstract Doc Link PMC2965573 Disease Relevance 1.20 Pain Relevance 0.03

General Comments

This test has worked.