Naive CD4 T-cell activation identifies MS patients having rapid transition to progressive MS

Evelyn Zastepa, Leslie Fitz-Gerald, Michael Hallett, Jack Antel, Amit Bar-Or, Sergio Baranzini, Yves Lapierre, David G Haegert, Evelyn Zastepa, Leslie Fitz-Gerald, Michael Hallett, Jack Antel, Amit Bar-Or, Sergio Baranzini, Yves Lapierre, David G Haegert

Abstract

Objective: Our objective was to determine whether altered naive CD4 T-cell biology contributes to development of disease progression in secondary progressive multiple sclerosis (SPMS).

Methods: We compared the naive CD4 T-cell gene expression profiles of 19 patients with SPMS and 14 healthy controls (HCs) using a whole-genome microarray approach. We analyzed surface protein expression of critical genes by flow cytometry after T-cell receptor (TCR) stimulation of naive CD4 T cells isolated from HCs and patients with SPMS.

Results: Hierarchical clustering segregated patients with SPMS into 2 subgroups: SP-1, which had a short duration of relapsing-remitting multiple sclerosis (MS), and SP-2, which had a long duration of relapsing-remitting MS. SP-1 patients upregulated numerous immune genes, including genes within TCR and toll-like receptor (TLR) signaling pathways. SP-2 patients showed immune gene downregulation in comparison with HCs. We identified an SP-1-specific transcriptional signature of 3 genes (TLR4, TLR2, and chemokine receptor 1), and these genes had higher surface protein expression in SP-1 than in SP-2. After TCR stimulation for 48 hours, only SP-1 showed a progressive linear increase in TLR2 and TLR4 protein expression.

Conclusions: Differences in naive CD4 T-cell biology, notably of TCR and TLR signaling pathways, identified patients with MS with more rapid conversion to secondary progression, a critical determinant of long-term disability in MS.

Figures

Figure 1. Hierarchical clustering segregated HCs from…
Figure 1. Hierarchical clustering segregated HCs from patients with SPMS and identified 2 SPMS subgroups
Unsupervised hierarchical clustering (Ward clustering method and Euclidean distance metric) of 1,439 genes having the greatest variance across all samples segregated HCs from patients with SPMS and identified 2 SPMS subgroups (SP-1 and SP-2). Patients with SPMS clustering with HCs are indicated in gray. HC = healthy control; SPMS = secondary progressive multiple sclerosis.
Figure 2. Transcriptional signature of 3 genes…
Figure 2. Transcriptional signature of 3 genes distinguishes SP-1 patients from other patients with MS and HCs
(A) SP-1 patients have higher expression of TLR2, TLR4, and CCR1 genes than either HCs or SP-2 patients. Mean fold change values are shown for SP-1 vs SP-2. (B) Hierarchical clustering based on the 3-gene list distinguishes SP-1 patients. Hierarchical clustering segregated SP-1 from HCs, SP-2, patients with stable RRMS, and patients with PPMS. HC = healthy control; MS = multiple sclerosis; PPMS = primary progressive MS; RRMS = relapsing-remitting MS.
Figure 3. Surface expression of 3-gene signature…
Figure 3. Surface expression of 3-gene signature and CD28
(A) Increased protein expression on freshly isolated naive CD4 T cells from SP-1. SP-1 patients had significantly higher percentage of TLR2, TLR4, and CCR1 than HCs and SP-2 patients (95% confidence interval not shown) and higher TLR4 MFI than HCs and SP-2 patients (p < 0.05, both comparisons). (B) TCR stimulation increased TLR2 surface expression in SP-1. At 48 hours, SP-1 had increased percentage of TLR2 and MFI (p < 0.05, both comparisons) compared with unstimulated cells, with a positive linear increase in percentage of TLR2 from 0 to 48 hours (p = 0.02). At 48 hours, SP-1 had higher percentage of TLR2 expression than HCs and SP-2 (p < 0.001, both comparisons) and higher TLR2 MFI than HCs and SP-2 (p < 0.05, both comparisons). (C) TCR stimulation increased TLR4 surface expression in SP-1 patients. The percentage of TLR4 increased in HCs (p < 0.05) and SP-2 (p < 0.01) from 0 to 24 hours, then decreased from 24 to 48 hours. In SP-1, percentage TLR4 showed a positive linear increase from 0 to 48 hours (p = 0.0025). At 48 hours, SP-1 had higher percentage of TLR4 than HCs or SP-2 (p < 0.01, both comparisons) and higher TLR4 MFI than HCs or SP-2 (p < 0.05, both comparisons). (D) CD28 surface expression. TCR stimulation for 24 hours decreased percentage CD28 (p < 0.001) and CD28 MFI (p < 0.05, not shown) in all groups, but at 24 hours, SP-1 had higher percentage CD28 than HCs (p < 0.05) or SP-2 (p < 0.01) plus higher CD28 MFI than HCs (p < 0.05, not shown). HC = healthy control; MFI = mean or median fluorescence intensity; TCR = T-cell receptor.

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Source: PubMed

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