Rheumatoid arthritis (RA) is a potentially sever and crippling disease with chronic smoldering immune responses causing tissue damage. Based on 2 immunological facts appear in RA patients, the persistent T lymphocytes immuno-activity & the imbalanced cytokine production by the inflammatory cells. It is likely that activated lymphocyte and Tumor Necrosis Factor-alpha (TNF-α) not only of major importance in RA pathogenesis but also they could be important in outcome prediction and possibly as therapeutic targets.
Hence this study tries to investigate the expression of activation markers on PBL, like the expression of CD45RA, CD45RO, CD54, and CD71 in RA patients, estimation of TNF-α serum levels in RA patients, Correlate the results of PBL-activation marker expression and serological estimation of TNF-α with different disease activity patterns, explore the possible role of activation markers and serum TNF-α level in prediction of the disease outcome. Initially Fourty six RA patients and 33 RA patients were reanalyzed after mean time 2 months, 7 patients with Osteo-arthritis (an attendant of Rheumatology clinic in Al-Kadhimyia hospital), and 10 apparently healthy individuals. The patients diagnosed by a physician and the questionnaire for each patient based on routine clinical and laboratory examination. Blood sample was taken from each subject and divided into 2 parts. The heparinized blood used for lymphocyte separation, smears were prepared, fixed
on charged slides, wrapped, and kept at -20°C until assayed. While, unheparinized blood allowed clotting, serum collected, and dispensed in aliquots and kept at -20°C until assayed. CD3, CD54, CD45RA, and CD45RO expression detected using Immunocytochemistry staining technique, CD71 detected using direct immunoflouresence staining technique, where as, the serum concentration of TNF-α was determined using Sandwich Enzyme-Linked Immunosorbent Assay (ELISA).The studied patients consist of 42 females and 4 males; there was a significant statistical difference in the frequency of patient in each sex group. The mean age of the patient was 48±12 and range from 25-66 years, most frequent age group was in 5th decade group (43%) from total patients. Most of the patients (80.4%) were presented with high disease activity. Rheumatoid factor (RF) was detected in 34 (74%) patient and all patients have high level of both rythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) serum level. Immunocytochemistry staining of CD3, CD54, CD45RO and immunoflourescence staining of CD71 revealed a highly statistical significant differences with higher mean percentage of expression in RA patients when compare with that of the healthy and patient (OA) control groups. While, CD45RA showed a lower percentage of cells expression among RA patients group with highly statistical significant difference when compared with that of the control groups. There was no statistical significant difference in mean percentage of cells that express CD3 and CD45RA between active and moderate disease activity groups. While, CD45RO showed a marginally statistical significant difference with lower percentage and CD54 and CD71 showed statistically significant
difference with higher expression in moderate disease activity group. There was also no statistically significant difference between initial and follow up group in expression of CD71, CD54, CD45RA, CD45RO and CD3. The sera of RA patients group revealed a statistically significant difference with highe concentration of TNF-α when compared with that of the control groups, and in active disease when compared with that of the moderate disease activity group, where as, no statistically significant difference between the initial an the follow-up groups. Serum TNF-α have a quite consistence and strong positive correlations with most of the clinical and laboratory disease activity indices, and unfortunately, there were no such correlations observed regarding PBL activation markers. Thus, we can depend on the serum TNF-α concentration as a laboratory parameter to define the degree of general disease activity (valid biomarker) for RA patients and using of it's blocker in treatment of patients especially those suffering from active disease.