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A Non-invasive Way to Isolate and Phenotype Cells from the Conjunctiva
Published on: July 5, 2017
Langerhans' cells in vernal keratoconjunctivitis express the costimulatory molecule B7-2 (CD86), but not B7-1 (CD80)
A M Abu-El-Asrar1, S A Al-Kharashi, S Al-Mansouri
1Department of Ophthalmology, College of Medicine, King Saud University, Riyadh, Saudi Arabia. abuasrar@KSU.edu.sa
Insights
Vernal keratoconjunctivitis (VKC) involves increased B7-2 expression on Langerhans' cells, interacting with CD28 on T cells. This pathway may drive the TH2 immune response, offering a target for new VKC therapies.
Area of Science:
- Immunology
- Ophthalmology
Background:
- Vernal keratoconjunctivitis (VKC) is linked to T-helper 2 (TH2) immune responses and elevated immunoglobulin E (IgE).
- Costimulatory molecule interactions, specifically B7 on antigen-presenting cells and CD28 on T cells, are crucial for T-cell activation and TH2 development.
Purpose of the Study:
- To investigate the expression of costimulatory molecules CD28, B7-1 (CD80), and B7-2 (CD86) in conjunctival biopsies.
- To compare the expression levels in patients with active VKC versus normal controls.
Main Methods:
- Immunohistochemical analysis of conjunctival biopsy specimens from 15 VKC patients and 8 controls.
- Utilized antibodies against CD28, B7-1, and B7-2.
- Sequential double immunohistochemistry to identify the phenotype of expressing inflammatory cells.
Main Results:
- B7-2 was significantly upregulated on mononuclear cells in VKC conjunctiva compared to controls (30.5 vs. 1.88, p < 0.001).
- B7-2 expression was more prevalent than B7-1 in VKC specimens (30.5 vs. 2.3, p < 0.001).
- CD28 was expressed on T lymphocytes, while B7-2 was primarily on CD1a+ Langerhans' cells in VKC.
Conclusions:
- B7-2 is prominently expressed by Langerhans' cells in VKC, suggesting a key role in the TH2 immune response.
- The B7-2/CD28 interaction is implicated in mediating the TH2 response in VKC.
- Targeting this costimulatory pathway presents a potential therapeutic strategy for VKC.
Purpose:
Vernal keratoconjunctivitis (VKC) is associated with T-helper 2 (TH2)-like cell response and increased immunoglobulin (Ig) E production. Recent studies have suggested that interactions between costimulatory molecules B7 on antigen-presenting cells and CD28 on T cells are critical for successful antigen presentation and the development of the TH2 immune response. The objective of this study was to examine the expression of costimulatory molecules CD28, B7-1 (CD80) and B7-2 (CD86) in conjunctival biopsies from patients with active VKC and normal controls.
Methods:
Conjunctival biopsy specimens from 15 subjects with active VKC, and 8 control subjects, were studied by immunohistochemical techniques using a panel of monoclonal and polyclonal antibodies directed against CD28, B7-1 and B7-2 molecules. The phenotype of inflammatory cells expressing costimulatory molecules was examined by sequential double immunohistochemistry.
Results:
In the normal conjunctiva, B7-2 was expressed on a few mononuclear cells in the epithelium and substantia propria in 5 of 8 specimens. There was no immunoreactivity for CD28 or B7-1. In VKC specimens, few B7-1+ mononuclear cells were noted in the substantia propria in 7 of 15 specimens. B7-2 was expressed on mononuclear cells in the epithelium and substantia propria in all specimens. Compared with normal controls, VKC specimens showed significantly more mononuclear cells expressing B7-2 (30.5+/-14.1 vs 1.88+/-2.5; p < 0.001). In VKC specimens, the numbers of mononuclear cells expressing B7-2 were significantly higher than the numbers of mononuclear cells expressing B7-1 (30.5+/-14.1 vs 2.3+/-3.1; p < 0.001). CD28 was expressed on mononuclear cells in the epithelium and substantia propria in 14 specimens. Colocalisation studies revealed that the majority of mononuclear cells expressing B7-2 were CD1a+ Langerhans' cells, and that the mononuclear cells expressing CD28 were CD3+ T lymphocytes.
Conclusions:
B7-2 is more widely and prominently expressed by Langerhans' cells compared with B7-1. The interaction of B7-2 with CD28 may mediate the development of the TH2 immune response in VKC. Thus the manipulation of this pathway could be an important target for the development of future therapies in VKC.
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