IL-7 and IL-7R Expression in Peripheral Blood Mononuclear Cells, Peripheral Blood Monocytes or Differentiated Macrophages of Rheumatoid Arthritis Patients With Active vs. Inactive Disease Treated With DMARD and/or CIMZIA

Blood samples will be collected from patients that have been diagnosed with RA based on ACR
classification criteria. The study will include 200 donors. The total number of subjects are
divided into two groups to yield a power of 95% at a type I error 5% level [determined based
on the preliminary data]. In the first group, 200 donors will be treated with methotrexate,
plaquenil and/or prednisone (DMARDs) that either achieve remission (DAS28<2.6) or do not
achieve remission (DAS28>2.6). 50 donor will be utilized as they respond to DMARDs and
achieve remission (DAS28<2.6) and 150 donors that do not respond to DMARDs will be
transferred to second group. In the second group, 150 donors will be treated with
methotrexate, plaquenil and/or prednisone and Cimzia® (provided to us by UCB).

In the first group of patients, blood samples will be obtained from RA patients treated with
Disease modifying anti rheumatic drugs (DMARDs) such as methotrexate, plaquenil and/or
prednisone that achieve remission (DAS28<2.6). The patients that achieve remission
(DAS28<2.6), blood will only be taken once at the patients routine visit.

The second group will consist of RA patients that did not respond to "DMARDs". These patients
will further receive (DMARDs) such as methotrexate, plaquenil and/or prednisone as well as
Cimzia® (provided to us by UCB) free of charge. Cimzia® is a FDA approved drug and is a
standard of care. Blood samples will be obtained from the patients treated with DMARDs
including methotrexate, plaquenil, and/or prednisone and Cimzia® (provided to us by UCB) that
have inactive remission (DAS28<2.6). In this group, blood samples will be collected onset of
the study as well as 3 and 6 months after treatment with Cimzia at patient's visit through
our collaboration with the aforementioned rheumatologists. Patients receiving intra-articular
steroid injections will be excluded from the study.

PB mononuclear cells will be isolated from RA whole blood and drawn into EDTA or CPT tubes
and isolated by Histopaque gradient centrifugation. Monocytes will be isolated from RA PB
mononuclear cells by negative selection (as shown in the preliminary data) and half of the
monocytes will be differentiated to macrophages for 7 days. The expression levels for IL-7
and IL-7R will be determined by real-time RT-PCR and FACS analysis.

In our statistical analysis, we will first perform a stratified analysis to evaluate the
differential expression levels in RA patients with active and inactive disease, controlling
for the type of treatment. Data analysis will be performed in collaboration with an UIC
Center for Clinical and Translational Science statistician. Specifically, we will perform the
comparison of IL-7 and IL-7R expression among RA patients with active (DAS28>2.6) vs.
inactive disease (DAS28<2.6) for DMARDs group (group 1). We will then perform a similar
comparison for the DMARDs and Cimzia® therapy group (group 2). The stratified analysis can
adjust for the potential confounding effect of treatment received and allows for the
detection of the potential differential relationships between expression levels of IL-7 or
IL-7R and disease status. We will also perform a pooled regression analysis in which the
expression logarithm of IL-7 or IL-7R from patients is regressed on the treatment group
indicator [DMARDs (group 1) versus on DMARDs plus Cimzia® therapy (group 2)] and disease
status (active or inactive disease) which would demonstrate the interaction between treatment
groups and disease activity. Such an analysis pools subjects from two treatment groups
together and can therefore increase the sample size, and hence potentially the power of
detecting the relationships between biomarkers and disease status. The RA samples will be
collected over a 2 year period and the data will be analyzed in the last year of the
proposal.

Inclusion Criteria:

1. Must meet 1987 Revised Criteria for the Classification of Rheumatoid Arthritis defined
as the diagnosis of the referring physician.

2. Persistent knee swelling (>ARA grade 2) for 2 weeks, and no recent intra-articular
corticosteroid injection.

3. Age 18 years and older.

4. Must be on Disease modifying anti rheumatic drugs (DMARDs) such as methotrexate,
plaquenil and/or prednisone.

Exclusion Criteria:

1. Patients having received intra-articular corticosteroid joint injection within the
last 2-4 weeks.

2. Patients with active systemic or joint infections.

3. Women who are pregnant (pregnancy status will be self-reported)

4. Patients under 18 years of age

5. Non-English speakers