Cytometric profiling in multiple sclerosis uncovers patient population structure and a reduction of CD8

Harvard Medical School/Partners Center for Genomics and Genetics, Brigham & Women's Hospital, 77 Avenue Louis Pasteur, NRB 168C, Boston, MA 02115, USA.
Brain (Impact Factor: 9.2). 06/2008; 131(Pt 7):1701-11. DOI: 10.1093/brain/awn118
Source: PubMed


As part of a biomarker discovery effort in peripheral blood, we acquired an immunological profile of cell-surface markers from healthy control and untreated subjects with relapsing-remitting MS (RRMS). Fresh blood from each subject was screened ex vivo using a panel of 50 fluorescently labelled monoclonal antibodies distributed amongst 56 pools of four antibodies each. From these 56 pools, we derived an immunological profile consisting of 1018 'features' for each subject in our analysis using a systematic gating strategy. These profiles were interrogated in an analysis with a screening phase (23 patients) and an extension phase (15 patients) to identify cell populations in peripheral blood whose frequency is altered in untreated RRMS subjects. A population of CD8(low)CD4(-) cells was identified as being reduced in frequency in untreated RRMS subjects (P = 0.0002), and this observation was confirmed in an independent sample of subjects from the Comprehensive Longitudinal Investigation of MS at the Brigham & Women's Hospital (P = 0.002). This reduction in the frequency of CD8(low)CD4(-) cells is also observed in 38 untreated subjects with a clinically isolated demyelination syndrome (CIS) (P = 0.0006). We also show that these differences may be due to a reduction in the CD8(low)CD56(+)CD3(-)CD4(-) subset of CD8(low) cells, which have a natural killer cell profile. Similarities between untreated CIS and RRMS subjects extend to broader immunological profiles: consensus clustering of our data suggests that there are three distinct populations of untreated RRMS subjects and that these distinct phenotypic categories are already present in our sample of untreated CIS subjects. Thus, our large-scale immunophenotyping approach has yielded robust evidence for a reduction of CD8(low)CD4(-) cells in both CIS and RRMS in the absence of treatment as well as suggestive evidence for the existence of immunologically distinct subsets of subjects with a demyelinating disease.

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    • "CD8low T cells are less cytotoxic than CD8high T cells, and they express IL-4, IL-10, and interferon-gamma [21]. CD8low lymphocytes, and in particular CD8low NK cells have been shown to be reduced in untreated patients with clinically isolated syndrome and MS [22]. Interestingly, MX appears to restore the frequency of CD8low T cells to the levels observed in healthy individuals (Fig. 1D), suggesting that this regulation may contribute to the efficacy of the treatment in MS. "
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    • "Nevertheless, studies using untreated MS patients have mostly detected deficits in NK cells function rather than differences in overall numbers between MS and controls. For instance, a reduction of an NK cell subtype, CD8lowCD56+CD3-CD4-, was observed in untreated, clinical isolated demyelination syndrome (CIS) and in relapse remitting MS (RRMS) patients, suggesting that this decrease in CD8low NK cells is an early event in demyelinating diseases [24]. The action of daclizumab (anti-IL2Rα) may restore to normal levels the CD8lowCD56+CD3-CD4- subset, this expansion correlating with decreased brain inflammation and decreased survival of activated T cells [21]. "
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    • "Reports of reduction of NK cell numbers and function in MS patients and evidence from experimental autoimmune encephalomyelitis (EAE) suggest a protective role of this population (Zhang et al., 1997; Kastrukoff et al., 2003; De Jager et al., 2008). However, this subset can also exert a detrimental role in the inflammatory process within the CNS (Winkler-Pickett et al., 2008). "
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