Analysis of cellular and humoral immune responses against cytomegalovirus in patients with autoimmune Addison’s disease

Abstract

BackgroundAutoimmune Addison’s disease (AAD) is caused by multiple genetic and environmental factors. Variants of genes encoding immunologically important proteins such as the HLA molecules are strongly associated with AAD, but any environmental risk factors have yet to be defined. We hypothesized that primary or reactivating infections with cytomegalovirus (CMV) could represent an environmental risk factor in AAD, and that CMV specific CD8+ T cell responses may be dysregulated, possibly leading to a suboptimal control of CMV. In particular, the objective was to assess the HLA-B8 restricted CD8+ T cell response to CMV since this HLA class I variant is a genetic risk factor for AAD.MethodsTo examine the CD8+ T cell response in detail, we analyzed the HLA-A2 and HLA-B8 restricted responses in AAD patients and healthy controls seropositive for CMV antibodies using HLA multimer technology, IFN-γ ELISpot and a CD107a based degranulation assay.ResultsNo differences between patients and controls were found in functions or frequencies of CMV-specific T cells, regardless if the analyses were performed ex vivo or after in vitro stimulation and expansion. However, individual patients showed signs of reactivating CMV infection correlating with poor CD8+ T cell responses to the virus, and a concomitant upregulation of interferon regulated genes in peripheral blood cells. Several recently diagnosed AAD patients also showed serological signs of ongoing primary CMV infection.ConclusionsCMV infection does not appear to be a major environmental risk factor in AAD, but may represent a precipitating factor in individual patients.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-016-0822-z) contains supplementary material, which is available to authorized users.

Increased infiltration and tolerised antigen-specific CD8+ TEM cells in tumor but not peripheral blood have no impact on survival of HCMV+ glioblastoma patients

“…Cells were stained with HCMV specific HLA-A2 or HLA-B8 restricted peptide dextramers ( Table 2 ) as described previously. 72 Fluorescent Minus One (FMO) controls were used for each channel, and lymphocytes were gated as indicated (Fig. S1A and Fig.…”

“…After 12 h of incubation at 37°C in 5% CO 2 , ELISpot membranes were developed, scanned and quantified as described previously. 72 IFNγ secretion was considered positive when the number of cells secreting IFNγ was ˃ 0 total cells.…”