Mechanism of Action of Ocrelizumab in Multiple Sclerosis
Conditions: Multiple Sclerosis, Immune System Diseases
Sponsor: University of Chicago

Jan 12, 2010  
 Immune Function in MS  AT Reder 
   
10681A 
NCT03344094 
 
Study Design/Rationale: 
Multiple sclerosis is an inflammatory disease that causes demyelination of the 
brain and spinal cord.  In MS, 20-60 year-old patients, women more than men, develop 
episodes of neurologic dysfunction that last days to weeks and then partially resolve.  
Damage is cumulative and eventually becomes progressive.  The immune system also 
evolves over time and becomes less sensitive to immune suppression.  Demyelinating 
variants of MS such as neuromyelitis optica and CNS Sjogren’s disease, have similar, but 
often more severe symptoms.  
 
We will be studying proteins and RNA in white blood cells, and how these 
change during therapy of MS and demyelinating variants.  White blood cells control 
some immune responses and may control the attacks, progression, and severity of MS.  
We will also analyze DNA polymorphisms of relevant genes identified in these studies.  
By studying these white blood cells, we hope to determine what causes MS and use this 
knowledge to design potential therapies for MS patients in the future. 
 
More specifically, assays will evaluate the following:  
 
Immune function: 
We will study immune function in multiple sclerosis, with 
assays of expression of costimulatory molecules and interferon receptors on the cell 
surface, also cytokine secretion, intracellular cytokines and signal transduction proteins, 
expression of mRNA for these proteins, NK function, and lymphocyte proliferation.  This 
information is intended to determine the mechanism of action of various therapeutic 
agents in MS (unknown at this time). 
 
Some examples of the cytokines to be investigated include interleukins, 
monokines, neuropeptides, interferons, TGF, and TNF; signal transduction molecules 
include JAK/STAT proteins, NF-kB, kinases, and phosphatases; costimulatory molecules 
include HLA-DR, B7.1, and B7.2. 
 
The particular assay is dependent on the mechanism of action of the agent being 
studied in MS.  All are variations on the same limited theme; all require a modest amount 
of blood; and all are subject to the same confidentiality arrangements.  As examples, we 
will use ELISA and RT-PCR to measure lymphocyte production of cytokines (such as 
IL-10 and IFN-gamma) protein and mRNA because of its relevance to immune function 
in MS patients before and after interferon therapy; use flow cytometry to measure 
lymphocyte B7.1 membrane expression for comparison to patients receiving Copaxone 
therapy, use Western blots and intracellular flow cytometry to measure STAT-l levels for 
comparison to patients receiving interferon therapy, and use RT-PCR, micro arrays and 
next generation RNA sequencing to measure RNA levels and exon use. 
 
Assays are done on blood from clinic population of MS patients and healthy age-
matched normal controls, and from clinical trial patients.  Blood from patients who are 
not part of trials, and also from patients who are in trials--before and after drug therapy, 
will be analyzed during the course of the study and compared to untreated patients and 
controls 

 
Clinic visits typically occur at 3-12 month intervals.  Trials of experimental or 
FDA-approved drugs last 1-3 years and typically require visits every 2-6 months. Blood 
will be drawn at regular clinic visits before therapy, at 1 & 3 mo, and at 1, 2, and 3 years 
after starting treatment, depending on duration of study.   Research will take place in J218 
and J220, SBRP. 
 
Kinetics: 
Some patients with stable and active and progressive MS 
will be tested while on drug treatment.  Here we will measure kinetic effects of 
immunomodulatory drugs.  In the latter patients, blood will be drawn before, and at 4, 18, 
and 42 hours or 5 days after drug administration.   
 
DNA: 
Blood from some patients with each form of MS and 
family members will be analyzed for genes linked to MS and controlling immune 
function.  In some cases, DNA may be obtained from buccal scraping or other body 
fluids. Multiple sclerosis is caused by interactions of multiple genes.  This is a broad 
screen of multiple genes to determine linkage to MS, and to determine responses to 
interferons.  Separate data on immune function, also without identifiers, will be compared 
to genes of interest.  These genes include, but aren't limited to, those regulating all 
aspects of immunity, cell survival, matrix metalloproteases and migration, kinases and 
phosphatases, and cell cycling, and pH regulation. 
 
In addition, we will assay methylation of DNA (circulating in plasma).  Some 
genes are methylated, a marker of their ability to be turned on that reflects cell regulation 
and death.  When cells die, they release DNA.  Sensitive assays can detect methylated 
forms of DNA from relevant genes (i.e., methylation-sensitive restriction enzyme 
digestion, then PCR). 
 
Al of these assays are directed at finding the immunologic cause of MS, how to predict 
exacerbations, and how to optimize MS therapy.