Frequency analysis of autoreactive T-helper 1 and 2 cells in bullous pemphigoid and pemphigus vulgaris by enzyme-linked immunospot assay

$48.00 plus tax (Refund Policy)

Download / Buy Article:

Abstract:

Background Bullous pemphigoid (BP) and pemphigus vulgaris (PV) are autoimmune bullous skin diseases mediated by autoantibodies against adhesion molecules of the skin. Previous studies have identified autoreactive T cells in patients with BP and PV, which may be critical in providing B-cell help for autoantibody production.

Objectives To evaluate the frequency of autoreactive T-helper (Th) 1 and Th2 cells in patients with BP (n = 7) or PV (n = 1) and in healthy controls (n = 11).

Methods In an enzyme-linked immunospot (ELISPOT) assay, microtitre plates were coated with antihuman interleukin (IL)-5 IgG or antihuman interferon (IFN)-γ IgG prior to culturing human peripheral blood lymphocytes (PBL) with BP180 or desmoglein (Dsg) 3 proteins for 7 days. Cytokine-producing autoreactive T cells were visualized as spot-forming units.

Results One BP patient with extensive blisters had 5·1 ± 1·5 (mean ± SD) BP180-reactive Th1 cells and 2·9 ± 1·5 Th2 cells per 105 PBL. In contrast, PBL from six BP patients in remission or on immunosuppressive therapy did not form IFN-γ- or IL-5-producing spots per ≤ 5 × 105 PBL. The patient with oral PV had 4·7 ± 2·4 Th1 cells and 3·0 ± 0·4 Th2 cells per 105 PBL and a vigorous PBL response to Dsg3. In addition, three of 10 controls had BP180-reactive Th1 (2·7–13·8 per 105 PBL) and Th2 (0·3–1·8 per 105 PBL) cells and one control had 9·0 ± 0·7 Th1 cells and 1·1 ± 0·8 Th2 cells per 105 PBL, with reactivity to Dsg3. ELISPOT reactivity correlated with 3H-thymidine incorporation in six of seven patients and controls with autoreactive T-cell responses.

Conclusions The ELISPOT assay seems to be promising for the quantitative and qualitative analysis of autoreactive T-cell responses in BP and PV.

Keywords: BP180; T-helper cells; autoreactivity; bullous pemphigoid; cytokines; enzyme-linked immunospot assay; pemphigus vulgaris

Document Type: Research Article

DOI: http://dx.doi.org/10.1046/j.1365-2133.2000.03901.x

Affiliations: 1: Department of Dermatology, RWTH Aachen, Germany 2: Department of Medicine, University of Cologne, Germany 3: Department of Dermatology, SUNY at Stony Brook, NY, U.S.A.

Publication date: December 1, 2000

Related content

Tools

Favourites

Share Content

Access Key

Free Content
Free content
New Content
New content
Open Access Content
Open access content
Subscribed Content
Subscribed content
Free Trial Content
Free trial content
Cookie Policy
X
Cookie Policy
ingentaconnect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more