Multiple Sclerosis Journal - October 2017 - 1480

Multiple Sclerosis Journal 23(11)
the Fc-fragment on antibodies opsonizing a target
cell. Only CD16-positive cells are capable of antibody-dependent cytotoxicity.5 CD56 dim NK cells
are less likely to proliferate after stimulation and are
thought to represent mature NK cells that have
evolved from CD56 bright cells.7-9 As mature cells,
they also express more classes and a higher amount of
inhibitory receptors on their surface to ensure that no
inappropriate lysis of target cells occurs.5 CD56 dim
numbers increase with age.8
CD56 bright NK cells
CD56 bright NK cells are thought to be more immune
regulatory due to being potent cytokine producers,
including the regulatory cytokines interleukin (IL)-10
and tumour necrosis factor (TNF)-beta.5,7,9 Due to
their homing receptors CCR7 and CD62L, they are
the predominant subset in lymph nodes.5 They are
also the predominant type of NK cells in the decidua
during pregnancy and are believed to be crucial in
preventing rejection of the unborn child.6 The CD56
bright NK cell population is increased in peripheral
blood during the last 3 months of pregnancy which
corresponds to the protective effect of pregnancy seen
on relapse frequency in MS, especially during the last
trimester.10,11
Initially believed to be less cytotoxic, the CD56 bright
subset can be equally cytolytic as the CD56 dim subset under the influence of IL2. Furthermore, CD56
bright NK cells as well as CD56 dim NK cells can kill
auto-reactive T cells in a contact-dependent manner in
response to stimulation of NK cells with IL2. 2,12,13
This interaction is particularly important for the new
treatment daclizumab, a monoclonal anti-CD25 antibody, where NK cells have been associated with a
highly favourable clinical outcome.1,2,12,14,15
As NK cells have been associated with superior clinical
outcome for daclizumab1,2,12,14,15 and beta-interferon
(IFN),16 we wanted to examine whether the role of NK
cells in disease control is independent of the therapeutic mode of action. In a retrospective study, we analysed NK cells and their subsets in peripheral blood of
relapsing-remitting multiple sclerosis (RRMS) patients
receiving the currently approved disease-modifying
treatments or no treatment and assessed the impact of
NK cell level on clinical outcomes.
Methods
Patients
We measured levels of lymphocyte subsets taken
from MS patients in our local MS centre between 14
1480

November 2008 and 14 March 2013. From a total of
148 patients, we selected 110 patients who were stable
with RRMS. If multiple suitable samples had been
taken, the sample representing the period of the longest
duration on treatment, or off treatment, was chosen.
Blood samples of subjects who were pregnant, breastfeeding, who had suffered a relapse or received steroids less than 28 days prior to sample collection were
excluded.
Patients were divided into active or non-active according to the No Evidence of Disease Activity (NEDA)
criteria17 over an observation period of up to 2.4 years.
Treatment response was defined according to NEDA
criteria in which the patient had to have no magnetic
resonance imaging (MRI) activity, no clinical relapses
and no Expanded Disability Status Scale (EDSS) progression. Secondary outcome parameters were MRI
activity, EDSS progression or relapse. MRI activity
was measured by new or enlarging lesions and/or
gadolinium enhancement. EDSS progression was
defined as increase in 1.0 or more on EDSS from a
baseline score of 1.0 or more, or an increase in 1.5 or
more from a baseline score of 0 that was sustained
over 12 weeks. Non-sustained progression at times of
relapse was not included.
Laboratory methods
Whole blood (ethylenediaminetetraacetic acid
(EDTA)) specimens were used for routine flow
cytometry analyses of lymphocyte subsets. Cells were
processed the same day; separation of CD56 into dim
and bright subsets was performed on on-list mode retrospectively. Specimens were stained with a Tetramer
multi-colour reagent against CD45-fluorescein isothiocyanate (FITC)/CD56-phycoerythrin (RD1)/CD19ECD/CD3-PC5. An aliquot (100 µL) of whole blood
was incubated with the Tetramer antibody (5 µL) for
10 minutes at room temperature and lysed using the
Q-Prep® system. A Navios flow cytometer was used
to acquire a minimum of 5000 lymphocytes using
CD45 versus side scatter for gating and then subgated using CD3 to obtain the CD3negative/CD56positive
subset (Figure 1). The absolute numbers for subsets
were calculated using the lymphocyte count obtained
from the same specimen on a LH780 Full Blood
Count analyser. Retrospective analyses were performed using Kaluza® analysis software to separate
the CD3negative/CD56 dim and CD56 bright subsets.
Kaluza analysis software is a Beckman Coulter
branded product for review of list-mode (or saved
data) data acquired from the Navios flow cytometer.
The person performing the analysis was blinded as to
the clinical parameters of patients.
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Table of Contents for the Digital Edition of Multiple Sclerosis Journal - October 2017

Contents
Multiple Sclerosis Journal - October 2017 - Cover1
Multiple Sclerosis Journal - October 2017 - Cover2
Multiple Sclerosis Journal - October 2017 - Contents
Multiple Sclerosis Journal - October 2017 - ii
Multiple Sclerosis Journal - October 2017 - iii
Multiple Sclerosis Journal - October 2017 - 1436
Multiple Sclerosis Journal - October 2017 - 1437
Multiple Sclerosis Journal - October 2017 - 1438
Multiple Sclerosis Journal - October 2017 - 1439
Multiple Sclerosis Journal - October 2017 - 1440
Multiple Sclerosis Journal - October 2017 - 1441
Multiple Sclerosis Journal - October 2017 - 1442
Multiple Sclerosis Journal - October 2017 - 1443
Multiple Sclerosis Journal - October 2017 - 1444
Multiple Sclerosis Journal - October 2017 - 1445
Multiple Sclerosis Journal - October 2017 - 1446
Multiple Sclerosis Journal - October 2017 - 1447
Multiple Sclerosis Journal - October 2017 - 1448
Multiple Sclerosis Journal - October 2017 - 1449
Multiple Sclerosis Journal - October 2017 - 1450
Multiple Sclerosis Journal - October 2017 - 1451
Multiple Sclerosis Journal - October 2017 - 1452
Multiple Sclerosis Journal - October 2017 - 1453
Multiple Sclerosis Journal - October 2017 - 1454
Multiple Sclerosis Journal - October 2017 - 1455
Multiple Sclerosis Journal - October 2017 - 1456
Multiple Sclerosis Journal - October 2017 - 1457
Multiple Sclerosis Journal - October 2017 - 1458
Multiple Sclerosis Journal - October 2017 - 1459
Multiple Sclerosis Journal - October 2017 - 1460
Multiple Sclerosis Journal - October 2017 - 1461
Multiple Sclerosis Journal - October 2017 - 1462
Multiple Sclerosis Journal - October 2017 - 1463
Multiple Sclerosis Journal - October 2017 - 1464
Multiple Sclerosis Journal - October 2017 - 1465
Multiple Sclerosis Journal - October 2017 - 1466
Multiple Sclerosis Journal - October 2017 - 1467
Multiple Sclerosis Journal - October 2017 - 1468
Multiple Sclerosis Journal - October 2017 - 1469
Multiple Sclerosis Journal - October 2017 - 1470
Multiple Sclerosis Journal - October 2017 - 1471
Multiple Sclerosis Journal - October 2017 - 1472
Multiple Sclerosis Journal - October 2017 - 1473
Multiple Sclerosis Journal - October 2017 - 1474
Multiple Sclerosis Journal - October 2017 - 1475
Multiple Sclerosis Journal - October 2017 - 1476
Multiple Sclerosis Journal - October 2017 - 1477
Multiple Sclerosis Journal - October 2017 - 1478
Multiple Sclerosis Journal - October 2017 - 1479
Multiple Sclerosis Journal - October 2017 - 1480
Multiple Sclerosis Journal - October 2017 - 1481
Multiple Sclerosis Journal - October 2017 - 1482
Multiple Sclerosis Journal - October 2017 - 1483
Multiple Sclerosis Journal - October 2017 - 1484
Multiple Sclerosis Journal - October 2017 - 1485
Multiple Sclerosis Journal - October 2017 - 1486
Multiple Sclerosis Journal - October 2017 - 1487
Multiple Sclerosis Journal - October 2017 - 1488
Multiple Sclerosis Journal - October 2017 - 1489
Multiple Sclerosis Journal - October 2017 - 1490
Multiple Sclerosis Journal - October 2017 - 1491
Multiple Sclerosis Journal - October 2017 - 1492
Multiple Sclerosis Journal - October 2017 - 1493
Multiple Sclerosis Journal - October 2017 - 1494
Multiple Sclerosis Journal - October 2017 - 1495
Multiple Sclerosis Journal - October 2017 - 1496
Multiple Sclerosis Journal - October 2017 - 1497
Multiple Sclerosis Journal - October 2017 - 1498
Multiple Sclerosis Journal - October 2017 - 1499
Multiple Sclerosis Journal - October 2017 - 1500
Multiple Sclerosis Journal - October 2017 - 1501
Multiple Sclerosis Journal - October 2017 - 1502
Multiple Sclerosis Journal - October 2017 - 1503
Multiple Sclerosis Journal - October 2017 - 1504
Multiple Sclerosis Journal - October 2017 - 1505
Multiple Sclerosis Journal - October 2017 - 1506
Multiple Sclerosis Journal - October 2017 - 1507
Multiple Sclerosis Journal - October 2017 - 1508
Multiple Sclerosis Journal - October 2017 - 1509
Multiple Sclerosis Journal - October 2017 - 1510
Multiple Sclerosis Journal - October 2017 - 1511
Multiple Sclerosis Journal - October 2017 - 1512
Multiple Sclerosis Journal - October 2017 - 1513
Multiple Sclerosis Journal - October 2017 - 1514
Multiple Sclerosis Journal - October 2017 - 1515
Multiple Sclerosis Journal - October 2017 - 1516
Multiple Sclerosis Journal - October 2017 - 1517
Multiple Sclerosis Journal - October 2017 - 1518
Multiple Sclerosis Journal - October 2017 - 1519
Multiple Sclerosis Journal - October 2017 - 1520
Multiple Sclerosis Journal - October 2017 - 1521
Multiple Sclerosis Journal - October 2017 - 1522
Multiple Sclerosis Journal - October 2017 - 1523
Multiple Sclerosis Journal - October 2017 - 1524
Multiple Sclerosis Journal - October 2017 - 1525
Multiple Sclerosis Journal - October 2017 - 1526
Multiple Sclerosis Journal - October 2017 - 1527
Multiple Sclerosis Journal - October 2017 - 1528
Multiple Sclerosis Journal - October 2017 - 1529
Multiple Sclerosis Journal - October 2017 - 1530
Multiple Sclerosis Journal - October 2017 - 1531
Multiple Sclerosis Journal - October 2017 - 1532
Multiple Sclerosis Journal - October 2017 - 1533
Multiple Sclerosis Journal - October 2017 - 1534
Multiple Sclerosis Journal - October 2017 - 1535
Multiple Sclerosis Journal - October 2017 - 1536
Multiple Sclerosis Journal - October 2017 - 1537
Multiple Sclerosis Journal - October 2017 - 1538
Multiple Sclerosis Journal - October 2017 - 1539
Multiple Sclerosis Journal - October 2017 - 1540
Multiple Sclerosis Journal - October 2017 - 1541
Multiple Sclerosis Journal - October 2017 - 1542
Multiple Sclerosis Journal - October 2017 - 1543
Multiple Sclerosis Journal - October 2017 - 1544
Multiple Sclerosis Journal - October 2017 - 1545
Multiple Sclerosis Journal - October 2017 - 1546
Multiple Sclerosis Journal - October 2017 - 1547
Multiple Sclerosis Journal - October 2017 - 1548
Multiple Sclerosis Journal - October 2017 - 1549
Multiple Sclerosis Journal - October 2017 - 1550
Multiple Sclerosis Journal - October 2017 - 1551
Multiple Sclerosis Journal - October 2017 - 1552
Multiple Sclerosis Journal - October 2017 - 1553
Multiple Sclerosis Journal - October 2017 - 1554
Multiple Sclerosis Journal - October 2017 - 1555
Multiple Sclerosis Journal - October 2017 - 1556
Multiple Sclerosis Journal - October 2017 - 1557
Multiple Sclerosis Journal - October 2017 - 1558
Multiple Sclerosis Journal - October 2017 - 1559
Multiple Sclerosis Journal - October 2017 - 1560
Multiple Sclerosis Journal - October 2017 - 1561
Multiple Sclerosis Journal - October 2017 - 1562
Multiple Sclerosis Journal - October 2017 - 1563
Multiple Sclerosis Journal - October 2017 - 1564
Multiple Sclerosis Journal - October 2017 - 1565
Multiple Sclerosis Journal - October 2017 - 1566
Multiple Sclerosis Journal - October 2017 - Cover3
Multiple Sclerosis Journal - October 2017 - Cover4
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