nn.neurology.org€¦ · web view2016/08/23 · comparable ifn-γ+ and il-17+ responses to...
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Supplemental Material for:
Distinct T Cell Signatures Define Subsets of Multiple Sclerosis Patients
Mark C. Johnson, Emily R. Pierson, Andrew J. Spieker, A. Scott Nielsen, Sylvia Posso,
Mariko Kita, Jane H. Buckner, and Joan M. Goverman
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figure e-1. Comparable IFN-γ+ and IL-17+ responses to stimulation with tetanus
toxoid and anti-CD3/28 antibodies are observed for HCs, brain-predominant and
spinal cord-predominant MS patients. PBMCs from HCs (n = 45), brain-predominant
(n = 29), and spinal cord-predominant (n = 16) MS patients were stimulated with either
(A) tetanus toxoid or (B) anti-CD3/CD28 antibodies and analyzed by ELISPOT.
ELISPOT = Enzyme-linked immunosorbent spot; HC = Healthy control subject; IFN =
interferon; IL = interleukin; MS = multiple sclerosis; PBMCs = Peripheral blood
mononuclear cells; SC = spinal cord
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figure e-2. MHC class II-, but not MHC class I-, blocking antibodies significantly
reduce MBP-specific IFN-γ+ frequencies. PBMCs from a subset of subjects were
analyzed for MBP-specific IFN-γ responses in samples that were pre-incubated with (A)
a MHC class I–blocking antibody or (B) MHC class II-blocking antibodies. Responses in
wells containing blocking antibodies and MBP (white bars) were directly compared to
wells incubated with MBP alone (black bars). *p < 0.05, ***p < 0.001, Two-way
ANOVA with Tukey’s multiple comparison test. Data represent mean ± SEM.
IFN = interferon; MBP = myelin basic protein; MHC = Major histocompatibility
complex; PBMCs = peripheral blood mononuclear cells
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figure e-3. Myelin antigen responses are not significantly different between HCs and
non-stratified MS patients. (A) PBMCs stimulated with MBP from HCs (n = 45) and
MS patients not segregated by neuroinflammatory pattern (n = 45) were analyzed for
IFN-γ+ and IL-17+ responses to MBP by ELISPOT. (B) MOG-specific IFN-γ+ and IL-17+
responses were calculated in a sub-set of HCs (n = 16) and combined MS patients (n =
29). ELISPOT = Enzyme-linked immunosorbent spot; HC = Healthy control subject; IFN
= interferon; IL = interleukin; MBP = myelin basic protein; MOG = myelin
oligodendrocyte glycoprotein; PBMCs = peripheral blood mononuclear cells; MS =
multiple sclerosis
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table e-1. HC and MS Patient Demographics
Group Healthy Controls(n = 45)
Brain-predominant(n =29)
Spinal cord-predominant
(n = 16)Female 35 (77.8%) 23 (79.3%) 13 (81.25%)
Male 10 (22.2%) 6 (20.7%) 3 (18.75%)
Age at draw
38.6 ± 9.5 40.6 ± 9.8 40.9 ± 8.2
EDSS score NA 0.7 ± 0.9 2.3 ± 2.5
EDSS = Expanded disability status scale; HC = healthy control subject; MS = multiple
sclerosis
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Glossary
AUC = Area under the curve
EDSS = Expanded disability status scale
ELISPOT = Enzyme-linked immunosorbent spot
EAE = Experimental autoimmune encephalomyelitis:
FPR = False positive rates
HCs = Healthy control subjects
IFN = Interferon
IL = Interleukin
MBP = Myelin basic protein
MHC = Major histocompatibility complex
MOG = Myelin oligodendrocyte glycoprotein
MS = Multiple sclerosis
PBMCs = Peripheral blood mononuclear cells
ROC = Receiver operating characteristic
RRMS = Relapsing-remitting multiple sclerosis:
TPR = True positive rates
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