الفهرس 
Acknowledgement
Rheumatoid arthritisOnly 14 pages are availabe for public view
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Abstract
This study involved 109 individuals divided to 3 groups, (group I) included 35 apparently healthy controls, (group II) included 35 patients diagnosed as diseased with Early Rheumatoid Arthritis and (group III): included 39 patients with late/chronic Rheumatoid Arthritis. A total number of 74 patients with RA were taken after approving of protocol by Local ethical committee (IBR 17200278) from November 2019 till November 2020. Patients recruited from clinic of Rheumatology and rehabilitation department, Assiut university Hospital as volunteers.
All patients in this study were subjected to the following:
1) Full history taking.
2) Clinical examination.
3) X-ray for affected joints and ultrasound examination if required.
4) Laboratory investigation:
• Complete blood counting
• Erythrocyte sedimentation rate (ESR)
• Liver function tests (Aspartate and Alanine transaminases (AST, ALT)) and Kidney function tests (serum creatinine & blood urea)
• Autoantibodies: for detection the values of rheumatoid factor
• Specific investigation (done for patients and Healthy control groups):
Detection of (CD19, CD24, CD25, CD38 & CD27) markers in peripheral blood samples by Flow Cytometer.
Determination of the serum level of IL 10 using Enzyme-Linked Immunosorbent Assay plate.
Determination of the serum level of anti-CP using ELISA plate.
History and clinical data demonstrate the following in the 3 studied groups; the age was significant older in the late RA patients group than in the control healthy group. The sex of the patients and healthy persons was highly significant different in-between the groups. The most of early and late RA patients were females. BMI mean was significant lower in early RA group than in late RA group and than in control group.
When compared between early and late RA concerning also the history and clinical data, we found that; deformity was significantly higher in late RA group than in early RA group. Grade of muscle weakness was significantly difference between early RA group and Late RA group. where the majority of early RA group was grade five and majority of late RA group was grade three. DAS-28 score was significantly higher in early RA group than in late RA group. Extra-articular manifestation was highly significantly higher in late RA group than in the early RA group. Hypertension and diabetes mellitus were significantly higher in late RA group than in early RA group. Hospitalization time mean was higher in late RA (3.8 times) than in early RA (0 times). Treatment modality was highly significant different between the early RA group and late RA group.
In this study we found that serum Anti-CP level was significantly lower in the control group than in late and in early RA groups.
Serum IL10 level reduces high significantly in late and early RA patients when compared with its level in healthy control group. Also, its level was moderate significantly higher in early RA than in late RA.
Breg cells or Transitional B cells% (particular CD25-Breg %) from B cells % was significantly higher in the early RA group than in the late RA group and in the control group. CD27-mature B cells % (particularly CD27-CD25-mature B cells%) from B cells % showed significantly lower percent in the late RA group than in the control group. CD27+mature B cells % (particularly CD27+CD25-mature B cells %) from B cells % was high significantly lower in early RA group and in late RA group than in the control group. But CD27+CD25+mature B cells % from B cells % significantly lower in early RA group and in late RA group than in the control group, while the % from CD27+mature B cells % was 37.3%in HC, 41.5% in early RA & 44.4%in late RA.
In treatment modalities (without treatment group, monotherapy, biological treatment, combined with MTX and without MTX) showed that IL10 level was moderate significantly lower in the group of patients under combined therapy without MTX than in the group under combined therapy with MTX, under biological treatment and without treatment.
The Breg cells % particularly CD25- Breg cells% not the CD25+ Breg% was significantly higher in the group without treatment than in the group under biological treatment and in the group under combined treatment without MTX.
The memory B cells % particularly CD25+memory B cells% not the CD25-memory B cells %, the CD27-CD25+ mature B cells %, the CD27+mature B cells % (both CD27+CD25+mature B cells % and CD27+CD25-mature B cells %) and CD25+PB cells % in the group under biological treatment was high significantly higher than in other groups.
The CD27-CD25-mature B cells% in the group under monotherapy was significantly higher than in the group under biological treatment and group under combined treatment without MTX. Beside that CD27+mature B cells % in the group under combined treatment without MTX was significantly higher than in the group without treatment.
PB cells % was significant higher in the group under combined therapy with MTX than in group without treatment and group under combined therapy without MTX. CD25-PB cells % was significantly higher in the group under combined therapy with MTX than in the group without therapy, in the group under combined therapy without MTX and in the group under biological treatment.
In early RA group, IL10 correlated positively with CD25-PB cells %. The CD25-Breg cells% and CD25+Breg cells % correlated positively with B cells %. But CD25+Breg cells % correlated highly significantly positive with CD27-CD25+mature B cells %.
The CD25+memory B cells % correlated significantly positive with RF level, DAS-28, Anti-CP level and significantly high with CD25-memory B cells %. That suggested that CD25+memory B cells % considers as marker of early RA only. It correlated significantly negative with CD25+PB cells%. CD25-memory B cells % correlated significantly positive with RF level and significantly negative with CD27-CD25+ mature B cells and B cells %.
The CD27-CD25-mature B cells % correlated significantly negative with RF, lymphocytes % and CD27+CD25+mature B cells %, but correlated positively with CD27+CD25- mature B cells %. CD27-CD25+mature B cells correlated significantly positive with B cells %.
The CD27+CD25-mature B cells % correlated significantly negative with DAS-28, Lymphocytes% and CD27-CD25+mature B cells %.
The CD25-PB cells % correlated significantly positive with CD25+PB cells %. B cells % correlated significantly negative with DAS-28.
In late RA IL10 correlated negatively with CD25-Breg cells % and DAS-28, positively with lymphocyte%, CD25-Memory B cell%, CD25+Memory B cell % and CD27+CD25+Mature B cell %.
The CD25-Breg cells% correlated significantly positive with DAS-28 and lymphocytes %. But it correlated significantly negative with the CD27+CD25- mature B cells%. This may relate to pathogenesis of the disease and the found of TNF. CD25+Breg cells% correlated significantly positive with CD25+memory B cells %, highly significantly with CD27-CD25+mature B cells %, CD27+CD25+mature B cells % and CD25-PB cells %. This may be because that this type of cells is the active phenotype that able to differentiate to other phenotypes of B cells.
CD25+memory B cells % correlated significantly positive with IL10 level, CD25-memory B cells %, CD27-CD25+mature B cells %, CD27+CD25-mature B cells and highly significantly with CD27+CD25+ mature B cells%. It correlated significantly negative with DAS-28. CD25-memory B cells % correlated significantly positive with IL10 level, Anti-CP level and CD27+CD25- mature B cells %.
CD27-CD25+mature B cells % correlated negatively with DAS-28. CD27+CD25+mature B cells % correlated significantly positive with CD27-CD25+mature B cells % and RF but significantly negative with DAS-28.CD27+CD25-mature B cells % correlated significantly negative with DAS-28 and significantly positive with IL10.
CD25-PB cells % correlated significantly positive with RF, CD27-CD25+mature B cells %, CD27+CD25+mature B cells % and CD25+PB cells %. CD25-PB cells % correlated significantly negative with B cells.