Role of the Immune System(s) in Progressive MS · 2018-04-02 · Emerging roles of: (i) Glial influences on neuronal physiology (ii) Immune:Glial interactions impacting neurons Astrocyte

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Role of the Immune System(s) in Progressive MS

Amit Bar-Or,, Professor, Neuroimmunology

Director, Experimental Therapeutics ProgramScientific Director, Clinical Research Unit

Montreal Neurological Institute, McGill University

CMSC Annual MeetingIndianapolis, May 2015

I have participated as a speaker at meetings sponsored by, received consulting fees, and/or grant support from:

Biogen Idec, Diogenix, Genentech, Sanofi-Genzyme EMD/Merck Serono, GSK, Mitsubishi Pharma, Novartis, Receptos, Roche,, Teva Neuroscience

Investigational and off label agents may be discussed.

Disclosures

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Concepts of inflammation and degeneration in MS: [Inflammation throughout MS spectrum (lesion axon loss microglial activation); divide into periph infilt vs CNS compartm]

How soon might it start – PD MS evidence (imaging, recent pathology - Bruck)?

Mechanisms: Adaptive (T and B specificity; ? Target/ pediatric Axoglial?) vs Innate (bystander; chronic microglial activation – more later)

Direct inflamma damage vs. indirect (impacting degeneration –eg OPC failed repair, astrocyte support, of OPC, neurons eg NK; exacerbate metabolic stress/demand/mitochondria, channelopathies etc

Single vs multi-hit (eg Jack’s Oligo model, then Moore et al direct inflamm/OPC or via astrocytes)

CNS compartmentalized – evidence late (Renyolds); early (Lucchinetti): microglia – Nat Neurosci

January 2002 February 2003 November 2004

Multi-focal (and diffuse) injury evolving over time

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Outline

Concepts of inflammation and degeneration in MS

Multi-hit model

Direct and Indirect immune effects

How soon might it start?

Inflammation invoked in Neurodegenerative Diseases

Emerging roles of inflammation impacting:

(i) Glial influences on neuronal physiology

(ii) Immune:Glial interactions impacting neurons

Adaptive and Innate immune responses

‘Degenerative’ and/orCNS-compartmentalized

Inflammatory InjuryPeripherally-mediatedInflammatory Injury

Imaging Course

Clinical Course

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Degenerative injury

Inflammatory Injury

Imaging Course

Clinical Course

Underlying BiologiesAdaptive

Innate

Degenerative injury

Inflammatory Injury

Imaging Course

Clinical Course


Innate

T cellsB cells

MyeloidNK cellsTissue

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CNS-Compartmentalizeddegeneration and inflammation

(Innate > Adaptive)Inflammation (Adaptive + Innate)Peripherally-Mediated

Imaging Course

Clinical Course


Innate

T cellsB cells

MyeloidNK cellsTissue

Imaging Course

Clinical Course

Underlying Biologies

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3

1. Inflammatory/Degenerative? Different inflammatory ‘flavors’?

2. Relation of Red to Blue? What if we eliminate blue?

3. What starts it all (‘the ‘chicken or the Egg’)?

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Inflammation and Degeneration in MS

“An ongoing quandary in MS is what causes the ultimate loss of axons and neurons that underlies disability progression.

Is there an evolving profile of [CNS] inflammation?

Is there another process of neurodegeneration?

If both, are these processes independent or somehow inter‐related?”

After Wolfgang BrÜck

Mechanisms that may underlie Progressive (‘non relapsing’) CNS injury

Inflammation: Perivascular lesion (Adaptive)

Focal and diffuse (Glial activation)

Meningeal inflammation (‘B-cell rich’)

Degeneration: ‘Toxins’: glutamate; O2, Nitrogen, Fe/HemeMitochondrial (demand outstrips supply…)

‘Functional channelopathies’

Neural-glial uncoupling; functional networks

Mechanisms above also likely contribute to limited repair

Loss of compensatory mechanisms + Ageing

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Inflammation: Adaptive (perivascular lesion?)

Innate (glial activation; focal +diffuse)

Immune:glial interactions; meningeal






T cells: Th1, Th2, Th9, Th17; Treg…

CNS-antigen directed immune responses

B cells: Plasmablasts/plasma cells, Abs …

Quite strongly implicated in RRMS

May also be implicated in progressive MS(Immune cells persisting in CNS? Antibodies?)

Yet: Relevant antigenic-specificities not fully elucidated (multiple reasons)…

Adaptive immune responses in MS

Regulatory (Breg), effector (Beff)

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Adaptive immune responses in MS

Cao et al Sci Trans Med, 2015

Cao et al Sci Trans Med, 2015

Abnormal responses of myelin-reactive T cells in MS

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ALS: Disease origin and progression in amyotrophic lateral sclerosis: an immunology perspective. Malaspina A, et al. Int Immunol. 2015.

AzD: Neuronal NLRP1 inflammasome activation regulates inflammatory interleukin‐1‐beta production and axonal degeneration. Kaushal V, et al. Cell Death Differ. 2015.

Chronic Epilepsy: NLRP1 inflammasome is activated in patients with TLE and contributes to neuronal pyroptosis. Tan CC, et al. J Neuroinflammation. 2015

MPS: Neuroinflammation, mitochondrial defects and neurodegeneration in mucopolysaccharidosis III type C mouse model. Martins C, et al. Brain. 2015.

Ceroid lipofuscinosis: An anti‐neuroinflammatory that targets dysregulated glia enhances efficacy of CNS‐directed gene therapy in infantile neuronal ceroid lipofuscinosis. Macauley SL, et al. J Neurosci. 2014.

Innate inflammation in multiple neurodegenerative diseases

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A BrainNet Europe gene expression microarray study. Durrenberger PF, et al. J Neural Transm. 2014.

Comparative genome-wide expression data (Illumina H-Ref 8)

AzD, ALS, HD, MS, PD, MS and Schizophrenia (n = 113 well-characterized post-mortem brain tissues).

Results: no dysregulated gene (passing QC) found across all 61 dysregulated genes shared when comparing 4+5 diseases)

Hints for common neuronal homeostatic, survival and synaptic plasticity pathways.

All diseases exhibited changes in several inflammation-related genes …role of the innate immune system in the pathogenesis and/or response to neurodegeneration.

Are there common mechanisms across diseases?

Outline


Cellular and molecular mechanisms by which inflammation may contribute to CNS degeneration



Emerging roles of:



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Outline


Cellular and molecular mechanisms by which inflammation may contribute to CNS degeneration



Emerging roles of:



Astrocyte and Microglia Biology: Novel functions and distinct subsets impacting OPC and Neurons

Molecular and functional microglia signaturesButovsky O, et al. Nat Neurosci. 2014

Glia-glia interactions: astrocyte and microglia effects on OPC during development and demyelinating disease.Clemente D et al. Front Cell Neurosci. 2013

Astrocytes-neuron signaling in brain.Tang F, et al. Nat Commun. 2014

Glial cells in progressive CNS injury and repair

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Several important neuronal–glial interactions recognized

Synapse loss is striking in neurodegenerative disease and

Glia have intimate roles in synapse physiology:

Astrocytes, OL, and microglia: All crucial and multifaceted roles in maintenance of synaptic function and excitability.

Neuroinflammation contribution to synapse loss may be primarily mediated by altered glial functions.

Question: how does Inflammation (aging, neuronal stress) impact on glia (eg activation) neurons/synapses

Glia: guardians, gluttons, or guides for maintenanceof neuronal connectivity? Jebelli, et al; Ann NY Acad Sci. 2015.

Microglial p53 activation is detrimental to neuronal synapses during activation-induced inflammation: Implications for neurodegeneration. Jebelli J, et al. Neurosci Lett. 2014.

Neurodegeneration by activation of the microglial complement-phagosome pathway. Bodea LG, et al. J Neurosci. 2014.

System xC- is a mediator of microglial function; its deletion slows symptoms in ALS mice. Mesci P, et al Brain. 2015.

SIRT1 deficiency in microglia contributes to cognitive decline in aging and neurodegeneration via epigenetic regulation of IL-1β. Cho SH, et al. J Neurosci. 2015.

A DAP12-Dependent signal promotes pro-inflammatory polarization in microglia following nerve injury and exacerbates degeneration of injured neurons. Kobayashi M, et al. Glia. 2015

Microglia: Inflammation glial response Neurodegeneration

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Adaptive Immune: T-cell-mediated regulation of neuroinflammation involved in neurodegenerative diseases. González H and Pacheco R. J Neuroinflammation. 2014.

Possible Mechanisms of Inflammation Degeneration

Innate immune: Inflammasome, TLR, Complement, Vesicles:The impact of single and pairwise Toll-like receptor activation on neuroinflammation and neurodegeneration.Rosenberger K, et al. J Neuroinflammation. 2014.

Versatility of the complement system in neuroinflammation, neurodegeneration and brain homeostasis. Orsini F. et al. Front Cell Neurosci. 2014.

Emerging roles of extracellular vesicles in the nervous system. Rajendran L, et al. J Neurosci. 2014.

The role of the immune system in neurodegenerative disorders: Adaptive or maladaptive?Doty KR, et al. Brain Res. 2014.

Neuroinflammation: Take the Bad with the Good?

Astrocyte-targeted production of IL-10 induces changes in microglial reactivity and reduces motor neuron death after facial nerve axotomy. Villacampa N, et al. Glia. 2015.

Protection of TGF-β1 against Neuroinflammation and Neurodegeneration in Aβ1-42-Induced Alzheimer's Disease Model Rats. Chen Jh et al. PLoS One. 2015.

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Microglia signature (gene expression and quantitative mass spectrometry) suggests ‘Maintainers of CNS quiescence’

TREM2 deficiency eliminates TREM2+ inflammatory macrophages and ameliorates pathology in Alzheimer's disease mouse models. Jay TR, et al. J Exp Med. 2015.

However:

TREM2 Lipid Sensing Sustains the Microglial desired Response in an Alzheimer's Disease Model. Wang Y, et al Cell. 2015

Adapted from Hohlfeld R et al. J Neuroimmunol. 2000;107:161-166.

The Two Sides of Inflammation

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Outline


Multi-hit model








Outline


Multi-hit model








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‘Multi-hit’ Model of Oligodendrocyte Injury

P53

• Inflammation• Ischemia• Infection • Trauma

p53 upregulation Sublethal injury

Death receptor upregulation

Fas

DR4/5

Death receptor triggering and apoptotic signalling Lethal injury

IFNgTNFa

T cellMicroglia

ROS

From Antel J. Clin Neurol Neurosurg. 2005

P53




Fas

DR4/5


IFNgTNFa

T cellMicroglia

ROS



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P53




Fas

DR4/5


IFNgTNFa

T cellMicroglia

ROS



From: Darlington P, et al J Neuropathol Exp Neurol. 2008

Human neurons live on ‘bed’ of astrocytes

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Immune cells can injure astrocytes with secondary injury to neurons

From: Darlington P, et al J Neuropathol Exp Neurol. 2008

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Reproduced with permission from Kutzelnigg A et al. Brain. 2005;128:2705-2712.

Subpial cortical demyelination in progressive MS

Kutzelnigg, A. et al. Brain 128:2705-2712; 2005

Meningeal inflammation in MS can be ‘B cell rich’

Howell OW, et al Brain. 2011

Uccelli et al, Trends Immunol 2005;

Corcione et al Autoimmunity Rev 2005

CD20

CD35

B C

CD20

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Cortical lesions: correlation with disease course, including disability and cognitive deficits (Howell OW, et al Brain. 2011 Sep;134(Pt 9):2755-71)

Microglial activation – identical gradient graded inflammatory process from pial surface in (Magliozzi R, et al Ann Neurol. 2010 Oct;68(4):477-93)

Subpial Cortical Lesions Exhibit Gradient of neuronal loss

Cortical layers

Magliozzi et al, 2010 Ann Neurol

Human Glial Cells/Neuroglia

OligodendrocyteAstrocyteMicroglia OPC

Trypsin

DNase

Percoll toremove myelin

Wash

5% MEM/DMEMMicroglia adherentOligos, others float

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‘State’ of myeloid cell: Balance betweenActivation and Quiescence/Inhibitory molecules

Microglia or

Macrophage

CD80

CCR7

HLA-DR

CX3CR1

SIRP1α

TREM-2

CD200R

Csfr

Adapted from Kierdorf et al., Frontiers in Cell Neuroscience 2013Hanane Touil

+ Unp B sup Activation/Quiescence markers (FACS +

qPCR)Purified Human

Microglia orMacrophage

Methods: effects of Breg and Beff products on microglia and macrophage Activation/Quiescence molecules

Hanane Touil

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p p p0

5

10

15

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mR

NA

Fo

ld C

ha

nge

HLA-DR

CCR7

0

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mR

NA

Fo

ld c

ha

ng

e

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mR

NA

F

old

Ch

an

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CD80

Human Microglia

Effects of Breg & Beff products on activation molecule expression by human macrophage and microglia

0

1

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mR

NA

Fo

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hang

e

p p p0.0

0.5

1.0

1.5

2.0

2.5

mR

NA

Fold

Change

HLA-DR

CCR7Human Macrophages

p p p0.0

0.5

1.0

1.5

2.0

mR

NA

Fo

ld C

ha

ng

e

CD80

Human Macrophages

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mR

NA

Fo

ld C

ha

ng

e

TREM-2

0

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mR

NA

Fo

ld C

han

ge

mCsfrp g p

0

10

20

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mR

NA

Fo

ld C

ha

ng

e

SIRP1α

Human Microglia

0.0

0.5

1.0

1.5

2.0

2.5

mR

NA

Fo

ld C

ha

ng

e

TREM-2p

0

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2

3

mR

NA

Fo

ld C

ha

ng

e

SIRP1α

0

1

2

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mR

NA

Fo

ld C

ha

ng

e

mCsfr

Effects of Breg & Beff products on quiescence/inhibitory molecule expression by human macrophage and microglia

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Outline


Multi-hit model








January 2002 February 2003 November 2004

Multi-focal (and diffuse) injury evolving over time

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Neurology. 2012 Jan 17;78(3):194-201

Summary: Inflammation in Progressive MSInflammation appears to be involved throughout MS spectrum; contributes in different ways in different CNS sub-compartments

Perivascular, diffuse, meningeal

Roles for both adaptive and innate; likely Innate > Adaptive

Important interface between inflammatory and degenerative mechanisms

Glial cells (microglia, astrocytes) as mediators of inflammation

Both direct and indirect injury mechanisms; ‘multiple hit’ model

‘Progressive disease biology’ may start earlier than we would like to think!

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Thanks to:

CIHR, MSSC, ITN/NIH, Wadsworth, CIHR/IHRT, MSSC Scientific Research Foundation

anti-CNS AntibodiesKevin O’Connor – YaleDavid Hafler – YaleEdgar Meinl – MunichChris Linington - Glasgow

Canadian PD ADS/MSBrenda Banwell – CHOP/SickKidsDoug Arnold - MNILouis Collins - MNIDessa Sadovnick – UBCRuth-Ann Marrie – U Manitoba

Glial-Immune interactionJack Antel – MNICraig Moore – Memorial UAlyson Fournier – MNITim Kennedy – MNIBob Lisak – Wayne stateValeria Ramaglia - Amsterdam

Canadian BMT/MSCT Group Mark Freedman - OttawaHarry Atkins - Ottawa

LaboratoryRui LiHanane TouilAyman RezkScott BellIna MexhitajLaila Al AlwanMalwina MencelLuke HealyMukanthu NyirendaAnshul AwasthiLeslie FitzgeraldMarie-Noelle BoivinFarzaneh Jalili

Experimental TherapeuticsSandeep VanamalaTrina JohnsonAda VilalobosBoli FanGregoire MorisseChahrazed Belabani

CIHR NET in AutoimmunityCiro Piccirillo - McGill Brenda Banwell - SickKidsPhil Sherman - SickKidsC. Polychronakos- McGillMark Silverberg – MSHAnne Griffiths - SickKids

Canadian B cell Team in MSJennifer Gommerman - U of TAlexandre Prat - CHUMSimon Fillatreau - Berlin

Virology/Vit D Raymond Tellier – EdmontonCarmen Yea – U of TReinhold Vieth – U of THeather Hanwell - U of T

‘Deep’ Immune MonitoringHeinz Wiendl – MunsterSven Meuth - Munster

Summary: Inflammation and Neurodegeneration

Valuable insights from comparing across diseases(Similarities and important differences)

Emerging roles of glial cells: neuronal integrity/function

Underscores importance of understanding both Neuronal:glial and Glial:Glial interactions

Neuroinflammation Glial state Neurodegeneration

Mechanisms may be starting at the very beginning…

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InflammatoryCNS Injury

A. DegenerativeCNS Injury

Bar-Or A. Advances in Neurology.;23:149-175, 2006

B. DegenerativeCNS Injury

DysregulatedImmune Response


DysregulatedImmune

Response

Two Views on Neuro-Immunology of MS

Two Views on Neuro-Immunology of MS


A. DegenerativeCNS Injury

B. DegenerativeCNS Injury

DysregulatedImmune Response


Bar-Or A. Advances in Neurology.;23:149-175, 2006

DysregulatedImmune

Response

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Approach: Blinded ‘unbiased’ proteomics of CSF obtained at time of initial episode of pediatric CNS inflammatory demyelination*

Subsequent ascertainment as MS (vs monophasic)

Hypothesis: Compact myelin antigens (MBP, PLP), traditionally considered disease-initiating, will be over-represented in CSF of children with MS than controls

Pediatric MS CSF may better reflect Early MS targets

* Canadian Pediatric Demyelinating Disease Study

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Less known features:

? Impact of ‘peripheral/relapsing inflammation’ on ‘CNS-compartmentalized inflammation’

Eg: peripheral immune cell responses impacting:- microglial activation- glia-glia interactions

? Impact of ‘CNS inflammation’ (whether peripheral or compartmentalized) on ‘non-inflammatory’ (‘degen’) aspects of disease:

- inflammation impacting mitochondrial function- Klaus Nave presentation at ISNI/Mahad

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Meningeal inflammation in MS is ‘B cell rich’ *

* Staining with aCD20 (B cell marker)(Howell OW, et al Brain. 2011 Sep;134(Pt 9):2755-7).

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Biologies Contributing to Central Nervous System Injury in MS

65RMS = relapsing multiple sclerosis; SPMS = secondary progressive MS; PPMS = primary progressive MS.Bar-Or A. Semin Neurol. 2008;28(1):29–45.

Typical clinical course of RMS and SPMS

Imaging course

Peripherally-initiatedinflammatory injury

CNS-compartmentalizeddegenerative and/orinflammatory injury

PPMS

B cells B cells?

Reprinted from Bar-Or A. Semin Neurol. 2008;28(1):29–45;Copyright 2008,Thieme Publishing Group.

Identification of a microglia signature by gene expression and quantitative mass spectrometry (AffyExon1 & MG400 chip)

Nat. Neurosci. 2014 Jan;17(1):131-43

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PRE-HSCT POST-HSCT

RE

LAP

SE

S D

UR

ING

2

YE

AR

PE

RIO

D

0

1

2

3

4

n=14

Immune ablation and autologous stem cell reconstitution*(BMT) essentially halts measures of new focal disease activity

* With M. Freedman, H. Atkins, D. Arnold, J. Chen, and the Canadian Collaborative BMT in MS Study Group

NU

MB

ER

OF

GD

+

LE

SIO

NS

PE

R S

CA

N

MONTHS POST-HSCT

BL

1

1 2 4 6 9 12 15 18 24

BL

2

0

5

10

15

20

25

MONTHS POST-HSCT

NU

MB

ER

OF

NE

W T

2 LE

SIO

NS

BL1-BL2 1 2 4 6 9 12 15 18 24

0

5

10

20

Darlington et al, Ann Neurol, 2013

Hawker K, et al. Ann Neurol. 2009;66:460–71.

Effects of B cell Depletion in PPMS

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Preplanned subgroup analyses

Effects of B cell Depletion in PPMS

Hawker K, et al. Ann Neurol. 2009;66:460–71.

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Presence of IgM OCB (M+) was associated strongly with presence of Gd+ lesions in PPMS

Villar et al Ann Neurol 2014

Gd+ lesions continued in M+ treated with placebo ..

No new Gd+ lesions in M+ treated with rituximab.

B cell depletion decreased IgM OCB but not IgG OCB:

Villar et al Ann Neurol 2014

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Peripheral Inflammation in PPMS (vs RRMS, SPMS)

Several studies – no clear convergence/conclusion

“We found unique autoantibody patterns that distinguished RRMS, secondary progressive (SPMS), and primary progressive (PPMS) MS from both healthy controls and other neurologic or autoimmune driven diseases including Alzheimer's disease, adrenoleukodystropy, and lupus erythematosus.”

Antigen microarrays identify unique serum autoantibody signatures in clinical and pathologic subtypes of multiple sclerosis. Quintana FJ; PNAS U S A. 2008

Lack of Replication; technology platforms; Cohort matching;(eg disease duration; Immune senescence)

Not convinced that relevant differences have been established between peripheral immune responses in PPMS and SPMS

Peripheral Inflammation in PPMS (vs RRMS, SPMS)

Several studies – no clear convergence/conclusion

“We found unique autoantibody patterns that distinguished RRMS, secondary progressive (SPMS), and primary progressive (PPMS) MS from both healthy controls and other neurologic or autoimmune driven diseases including Alzheimer's disease, adrenoleukodystropy, and lupus erythematosus.”

Antigen microarrays identify unique serum autoantibody signatures in clinical and pathologic subtypes of multiple sclerosis. Quintana FJ; PNAS U S A. 2008

Lack of Replication; technology platforms; Cohort matching;(eg disease duration; Immune senescence)

Not convinced that relevant differences have been established between peripheral immune responses in PPMS and SPMS

Documents

Role of the Immune System(s) in Progressive MS · 2018-04-02 · Emerging roles of: (i) Glial influences on neuronal physiology (ii) Immune:Glial interactions impacting neurons Astrocyte