Multiple Sclerosis: From Bedside to

Bench and Back Again

Jeri-Anne Lyons, Ph.D.

Department of Health Sciences
University of Wisconsin-Milwaukee
 Central Nervous System Diseases
& Neurodegeneration

 Multiple Sclerosis  Hallmarks of disease

 Immune-mediated  Periventricular lesions
 Destruction of the  Mononuclear
myelin sheath & axons inflammatory cells +/-
in the central nervous antibodies
system  Demyelination
 Signs and symptoms  Relative axon sparing
depend on the
location, size, and
 Astrocyte hypertrophy
number of lesions  Cervical spinal cord
(“plaques”) disproportionately
 Four main clinical involved
subtypes
 Demographics of MS

 Average age of onset 15 to 45 years1

 Gender ~70% women2
 Geography Incidence increases with distance from
equator in both directions 3

 Incidence 8,500 to 10,000 new cases per year 2

 Prevalence 0.1% of US population (350,000) 2

 Race Caucasians > other ethnic groups
1. Anderson DW et al. Ann Neurol. 1992;31:333-336.
2. Jacobsen DL et al. Clin Immunol Immunopathol. 1997;84:223-243.
3. Hauser SL. Harrison’s Principles of Internal Medicine. 1994.
 Etiology of Multiple Sclerosis

Environmental Factors

Familial tendency

Susceptibility
genes

infectious agent (s)

Autoimmune
reaction

Adams, Victor. Principles of Neurology. 5th ed. 1993;776-798

MS Lesions
 Disease Course in MS
(55%)* (10%)*

(30%)* (5%)*

Adapted from Lublin FD, Reingold SC. Neurology. 1996;46:907-911; * Jacobs et al. Mult Scler. 1999;5:369-376.
Clinical Course vs. Pathology
Disease Severity

Time (months/years)
Diagnostic Tests

 Magnetic resonance imaging (Brain and Spinal cord-

especially cervical cord)
 Analysis of spinal fluid
 Evoked potentials (visual, brainstem/auditory,
somatosensory) – used less often
MRI of normal vs. MS Brains
 Evidence for Immune System
Involvement in MS
 Inflammation
 CSF humoral dysimmune phenomena
 Peripheral blood dysimmune phenomena
 HLA (major histocompatibility) types associated with
susceptibility
 Female predominance (3:2)
 Response to immunomodulatory agents
 Animal models

Neuroimmunology Section, Washington University

 Antibody Abnormalities in CSF with
MS
 Increased CSF Index
IgG CSF/albumin CSF
IgG serum/albumin serum

 Increased IgM and IgG

synthesis rate
 Oligoclonal bands

**http://library.med.utah.edu/kw/ms/mml/ms_oligoclonal.html
John Rose, M.D., Maria Houtchens
Send in the murines

Research in an animal model

Experimental Autoimmune
Encephalomyelitis (EAE)
 Inflammatory demyelinating disease of the CNS
 Primary animal model for MS
 Autoimmune response to myelin proteins mediated by
CD4+ Th1 T cells
 Induced by active immunization with myelin proteins
 Myelin Basic Protein (MBP)
 Proteolipid Protein (PLP)
 Myelin Oligodendrocyte Glycoprotein (MOG)
 Disease characterized by ascending paralysis beginning
approximately 14dpi or 10dpt
 Pathology predominantly observed in lumbar region of
the spinal cord
EAE in Various Species and Strains
Lewis Rats MBP
B10.PL mice (MBPAc1-11)

SJL mice PLP (PLP139-151)

MBP (MBP89-106)
MOG (MOG92-
Marmoset 106)
(Callithrix MOG (MOG24-46)
jacchus)
C57BL/6 mice MOG (MOG35-55)
BALB/c mice PLP (PLP180-199)
EAE Grading Scale

0 Healthy
1 Loss of tail tone
2 Hind limb weakness
3 Single hind limb paralysis
4 Both hind limbs paralyzed
5 Dead or moribund
 rMOG vs. MOG35-55 induced EAE in WT vs. B
cell-/- mice

MOG(35-55) immunization rMOG immunization

5 WT
5

Clinical Score
4 B cell-/- 4
Clinical Score

3 3
2 2
1
1
0
0
0

27

35
17
24

32

39
42
46

0 10 20 30 40
Day Post Immunization
Day Post Immunization
Reconstitution of EAE in B cell-/- mice

14-16d later,
Immunize collect serum (Ab)
w/ Ag and remove
spleens (for B cells)
WT B6 mouse

Immunize
w/ Ag
Observe for EAE B cell-/- B6 mouse
Clinical EAE in B cell-/- mice: rMOG B cells

5
Mean Clinical Score

WT Controls
4
rMOG CD19+ B cell recipients
3
rMOG CD19+ CD80+
2 B cell recipients

1 B cell-/- controls

0
0 10 20 30 40 50
day post immunization
Clinical EAE in B cell-/- mice: rMOG serum Ab

5
mean clinical score

4 WT B6
B cell-/-
3 rMOG serum recipients
2 B cell-/- control
received serum 31dpi
1

0
0 10 20 30 40 50
received
serum:
day post immunization
 BcR

Bind
B cell Ag
BcR Plasma “Other
Cell Functions”
Antibody
B cell
“Activated”
other immune functions

Potential Roles for B cells in EAE/MS

Advantageous Harmful
B cells • Alter T cell • Initiate T cell
response response
(Th1 Th2) (Ag proc./presentation;
costimulation)
Antibody •Remyelination • Recruit Cells to CNS
• Demyelination
• Opsonization
• C’ activation
Taking it back to the clinic
Rituximab
 Mouse/human chimeric antibody vs. CD20
 Targets pre-B cells & mature B cells
 Spares stem cells, plasma cells
 Complement-mediated lysis

 FDA-approved for treatment

 1997: B cell lymphoma

 2006: Rheumatoid Arthritis

 Early studies suggest successful as monotherapy in

Relapsing-Remitting MS
Patient Population
Treatment Protocol

Rituximab Study Protocol

-8w -4w 0w 12w 16w 20w 24w 28w 32w 52w
4w 4w 12w 4w 4w 4w 4w 4w 20w

Rituximab
MRI
MSFC
CSF
Antibodies
Results
 Prolonged depletion of B cells
 Decreased MRI activity
 Some clinical effect observed
 No effect on CSF immunoglobulins
 Synthesis or oligoclonal bands
Surprising Results
Summary
 Multiple sclerosis is an autoimmune
demyelinating disease
 Disease is initiated by myelin-reactive T cells
 B cells and antibody are important to disease
pathology
 Targeting B cells may be an effective therapy
in those patients not responding to current
therapy
Acknowledgements
 Washington University
 Dr. Anne Cross
 Dr. Rob Naismuth

 Dr. Laura Piccio

 Michael Ramsbottom