OBJECTIVETo explore the pathogenesis of rheumatoid arthritis (RA) by studying the expression of T cell receptors (TCRs).

METHODST cell receptor Vbeta (TCR Vbeta) gene usage and expression were analyzed from synovial membrane and peripheral blood of 8 RA patients, 2 osteoarthritis patients and 2 accident amputees. The complementary determining region 3 (CDR3) of 25 TCR Vbeta subfamily genes in unselected T cell populations were amplified semi-quantitatively by reverse transcription-polymerase chain reaction (RT-PCR). The products were further studied by genescan for frequency of Vbeta usage.

RESULTSThe numbers of Vbeta subfamilies expressed by T cells from RA peripheral blood and synovial membrane were not significantly restricted. More importantly, biased Vbeta gene expression in RA synovium was observed and Vbeta6, Vbeta17, and Vbeta22 genes were the predominant subfamilies. It was noteworthy that the expression of Vbeta17 in RA synovium was significantly increased.

CONCLUSIONOur data were consistent with the hypothesis that several antigen or superantigen-driven processes may be involved in the pathogenesis of RA.

Arthritis, Rheumatoid ; genetics ; immunology ; Genes, T-Cell Receptor beta ; Humans ; Synovial Membrane ; metabolism ; T-Lymphocytes ; immunology

Objective To identify the potential long non-coding RNA (lncRNA) expressed in rheumatoid arthritis (RA) synovium key to RA onset and investigate its association with immune cell infiltration. Methods RA synovium data were downloaded from the GEO database and normalized. The lncRNAs key to RA onset were identified using multiple machine learning methods. Infiltration of 22 immune cell populations in RA synovium was measured by cell-type identification by estimating relative subsets of RNA transcripts (CIBER-SORT). The relationship between the key lncRNA and infiltrating immune cells was analyzed. Finally, real-time quantitative PCR was applied to validate the expression of the key lncRNA in RA synovial cells. Results lncRNA human leukocyte antigen complex P5(HCP5) was identified as the key lncRNA associated with RA onset. Infiltration analysis revealed increased abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages while decreased abundance of M2 macrophages in RA synovial tissue. Correlation analysis demonstrated that the lncRNA HCP5 expression was positively associated with the infiltration abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages in RA synovial tissue. Furthermore,the expression of lncRNA HCP5 in RA synovial cells was up-regulated. Conclusion lncRNA HCP5 expression is up-regulated in RA synovial tissue and potentially associated with immune cells infiltration.
Humans ; Arthritis, Rheumatoid ; CD8-Positive T-Lymphocytes ; HLA Antigens/metabolism* ; RNA, Long Noncoding/metabolism* ; Synovial Membrane/metabolism*

Rheumatoid arthritis is a chronic inflammatory systemic disease of young or middle aged adults, characterized by destructive and proliferative changes in synovial membrane, periarticular structure, skeletal muscle and perineural sheaths. Eventually, joints are destroied, ankylosed and deformed. Moderate anemia is occured frequently in rheumatoid arthritis patients, and these patients show abnormalities of iron metabolism such as lower serum iron concentration and occurance and distribution of iron in the synovial membrane. It has been suggested that the mechanism of iron deposits in rheumatoid arthritis is continuous oozing of blood from vascular granulation tissue into the synovial cavity and this lead the patient to anemia. We collected samples from serum and knee joint fluid in 21 cases of rheumatoid arthritis for chemical estimation of the ferritin concentration by radioimmunoassay, and compared with that of osteoarthritis. The following results were obtained: 1. The mean hemoglobin concentration(11.4gm %) in rheumatoid arthritis was lower than osteoarthritis(13.4gm %). 2. The mean serum ferritin concentration(118. 4ng/ml) in rheumatoid arthritis was lower than osteoarthritis(135. 6ng/ml), on the contrary in synovial fluid rheumatoid arthritis(279.8ng/ml) showed higher than osteoarthritis(190. 4ng/ml). 3. The mean ratio of synovial fluid ferritin on serum ferritin was 2. 36 in rheumatoid arthritis, in contrast with l. 4 in osteoarthritis. There was significant correlation between the ferritin concentration in synovial fluid and serum. 4. Serial check of ferritin concentration in synovial fluid during treatment would be thought meaningful criteria for determination of progress.and effectiveness of treatment.
Adult ; Anemia ; Arthritis, Rheumatoid ; Ferritins ; Granulation Tissue ; Humans ; Iron ; Joints ; Knee Joint ; Metabolism ; Middle Aged ; Muscle, Skeletal ; Osteoarthritis ; Radioimmunoassay ; Synovial Fluid ; Synovial Membrane

OBJECTIVE: To examine the expression of chemokine receptor CCR10 on monocytes/macrophages in the joints of patients with rheumatoid arthritis (RA), and to investigate the role of chemokine CCL28 and its receptor CCR10 in the migration of RA monocytes and its mechanism. METHODS: The expression of CCR10 in synovial tissues from 8 RA patients, 4 osteoarthritis (OA) patients, and 4 normal controls was analyzed by immunohistochemistry, and cell staining was scored on a 0-5 scales. Flow cytometry was used to measure the percentage of CCR10 positive cells in CD14⁺ monocytes from peripheral blood of 26 RA patients and 20 healthy controls, as well as from synovial fluid of 15 RA patients. The chemotactic migration of monocytes from RA patients and healthy controls in response to CCL28 was evaluated using an in vitro Transwell system. Western blotting was conducted to assess phosphorylation of the extracellular signal-regulated kinase (ERK) and protein kinase B (Akt) pathways in RA monocytes upon CCL28 treatment. RESULTS: CCR10 was predominantly expressed in RA synovial lining cells and sublining macrophages, endothelial cells, and lymphocytes. CCR10 expression was significantly increased on lining cells and sublining macrophages in RA synovial tissue compared with OA and normal synovial tissue (both P < 0.01). The patients with RA had markedly elevated expression of CCR10 on peripheral blood CD14⁺ monocytes compared with the healthy controls [(15.6±3.0)% vs. (7.7±3.8)%, P < 0.01]. CCR10 expression on synovial fluid monocytes from the RA patients was (32.0±15.0)%, which was significantly higher than that on RA peripheral blood monocytes (P < 0.01). In vitro, CCL28 caused significant migration of CD14⁺ monocytes from peripheral blood of the RA patients and the healthy controls at concentrations ranging from 10-100 μg/L (all P < 0.01). The presence of neutralizing antibody to CCR10 greatly suppressed CCL28-driven chemotaxis of RA monocytes (P < 0.01). Stimulation of RA monocytes with CCL28 induced a remarkable increase in phosphorylation of ERK and Akt (both P < 0.05). ERK inhibitor (U0126) and phosphatidylinositol 3-kinase (PI3K) inhibitor (LY294002) strongly reduced the migration of RA monocytes in response to CCL28 (both P < 0.01). CONCLUSION RA patients had increased CCR10 expression on peripheral blood, synovial fluid, and synovial tissue monocytes/macrophages. CCL28 ligation to CCR10 promoted RA monocyte migration through activation of the ERK and PI3K/Akt signaling pathways. The CCL28-CCR10 pathway could participate in monocyte recruitment into RA joints, thereby contributing to synovial inflammation and bone destruction.
Humans ; Monocytes/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Endothelial Cells/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Arthritis, Rheumatoid ; Synovial Membrane ; Chemokines, CC/metabolism* ; Synovial Fluid ; Osteoarthritis ; Receptors, CCR10/metabolism*

OBJECTIVETo investigate the role of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in the pathogenesis and progression of Kashin-Beck disease (KBD) and primary osteoarthritis (OA).

METHODSThe synovium and synovial fluid of the knee joint were collected from 20 adult patients with KBD, 18 with OA and 19 with meniscus injury (controls). The expression of IL-1beta and TNF-alpha in the synovium were analyzed by immunohistochemistry staining, and the levels of IL-1beta and TNF-alpha in the synovial fluid were determined by enzyme-linked immunosorbent assay.

RESULTIL-1beta and TNF-alpha expressions in the synovium and their levels in the synovial fluid were significantly higher in patients with KBD and OA than in patients with meniscus injury (P<0.05), but comparable between KBD and OA groups (P>0.05).

CONCLUSIONIL-1beta and TNF-alpha may play an important role in the pathogenesis and progression of KBD and OA.

Adult ; Aged ; Cartilage, Articular ; metabolism ; Endemic Diseases ; Female ; Humans ; Interleukin-1beta ; metabolism ; Male ; Middle Aged ; Osteoarthritis ; etiology ; metabolism ; Synovial Fluid ; metabolism ; Synovial Membrane ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Tatrate resistant acid phosphatase (TRAP) has been widely used as histochemical marker to identify osteoclast in studies of bone metabolism. However, the value of TRAP as an osteoclast marker is still in discussion. Authors isolated and characterized the cells from synovium of 6 patients of sero-positive rheumatoid arthritis and 4 patients of osteoarthritis, and observed the activity of acid phosphatase (AP) and TRAP. The activity of TRAP was negative in cell cultures in early phase, but the activity of TRAP was increased with time, and at one week the activity of TRAP was as strong as that of AP. If the cultured tissue contained bone, there were observed TRAP positive mononuclear cells and giant cells even in early phase of cultures (1 day, 3 day), and the phenotype of these cells were same as that of osteoclasts and osteoclast precursors by immunocytochemistry. In conclusion, the activity of TRAP was positive in cultured macrophage. TRAP is not a specific marker for osteoclast, and its use for the identification of osteoclast seems meaningful only in the early stage of cell culture.
Acid Phosphatase ; Arthritis, Rheumatoid ; Cell Culture Techniques ; Giant Cells ; Humans ; Immunohistochemistry ; Macrophages ; Metabolism ; Osteoarthritis ; Osteoclasts ; Phenotype ; Synovial Membrane

OBJECTIVETo investigate the role of natural killer-22 (NK-22) cells in the synovial fluid in the proliferation of fibroblast-like synoviocytes (FLS) in patients with rheumatoid arthritis (RA) and explore the possible signal pathway involved.

METHODSNK-22 cells in the SF of RA patients were sorted by flow cytometry. NK-22 cells were cultured for two weeks and the purity was detected by flow cytometry before stimulation with 20 ng/ml phorbol 12-myristate 13-acetate and 0.5 µmol/L ionomycin for 4 h. The level of interleukin-22 (IL-22) in the culture medium supernatant was then measured with ELISA. The proliferation of FLS in the presence of the culture supernatant of NK-22 cells was assessed with MTT assay at 24, 48 and 72 h, and the effect of IL-22 antibody on FLS proliferation was also observed. Real-time PCR and Western blotting were used to detect Stat3 mRNA and p-Stat3 protein levels, respectively, in the FLS exposed to rhIL-22 and AG490.

RESULTSNK-22 cells were successfully sorted by flow cytometry with a purity exceeding 90%. The levels of IL-22 in the supernatant of NKp44(+)NK cell culture averaged 1273.42∓254.48 pg/ml. The FLS proliferated rapidly 24, 48, and 72 h after the addition of culture supernatant of NK-22 cells (P<0.05). IL-22 antibody obviously inhibited the proliferation of FLS induced by NK-22 cell culture supernatant (P<0.05). Exposure of the FLS to rhIL-22 obviously increased cellular Stat3 expression levels, which were significantly lowered by the addition of AG490 (P<0.05).

CONCLUSIONNK-22 cells in the SF of RA patients can produce high concentrations of IL-22 to promote the proliferation of FLS through the STAT3 signal pathway.

Arthritis, Rheumatoid ; metabolism ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; cytology ; Humans ; Interleukins ; metabolism ; Killer Cells, Natural ; cytology ; metabolism ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Synovial Fluid ; cytology ; Synovial Membrane ; cytology

OBJECTIVETo explore the effect of Bushen Gujin Recipe (BGR) on serum and synovial expression of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α) in knee osteoarthritis (KOA) model rabbits.

METHODSTotally 36 8-month-old healthy male New Zealand white rabbits were randomly divided into 4 groups, i.e., the normal control group, the model group, the Western medicine group (Meloxicam, at the daily dose of 6 mg/kg), and the TCM group (BGR, at the daily dose of 53 g/kg), 9 in each group. Modeling was performed in all rabbits except those in the normal control group by using Hulth A method. All medication was performed for 8 consecutive weeks. Contents of IL-1 and TNF-α were detected using ELISA from serum, partial synovial tissue of the front knee joint, cartilage and subchondral bone of the medial femoral condyle.

RESULTSThe joint space became narrowed in the Western medicine group, ranging between the model group and the TCM group. The articular surface was rough with obvious osteophytes. The joint space was slightly narrower in the TCM group; the articular surface was slightly rough with mild osteophytes. Compared with the normal control group, contents of IL-1 and TNF-α in serum and synovial increased in the model group (P < 0.01). Compared with the model group, contents of IL-1 and TNF-α in serum and the synovial fluid decreased in the two treatment groups (P < 0.01). There was no statistical difference in contents of IL-1 and TNF-α between the Western medicine group and the TCM group.

CONCLUSIONBGR promoted the synthesis of cartilage matrix and carti- lage repair through inhibiting the secretion of IL-1 and TNF-α, and prolonging cartilage degeneration.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Interleukin-1 ; metabolism ; Knee Joint ; Male ; Osteoarthritis, Knee ; drug therapy ; metabolism ; Rabbits ; Synovial Fluid ; Synovial Membrane ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVES: Mutation of p53 may play a role in manifestation of rheumatoid arthritis synovium, but several studies on p53 expression in synovial tissues of rheumatoid arthritis showed conflicting results. We investigated the amount and pattern of p53 positive cells in rheumatoid arthritis synovium, in comparison with osteoarthritis synovium, by using immunohistochemistry with two other monoclonal antibodies for p53. METHODS: Synovial tissues from 9 patients with rheumatoid arthritis and 5 patients with osteoarthritis were examined for p53 expression by immunohistochemistry with 2 monoclonal antibodies for p53, DO-1 and DO-7. Histologic features of inflammation were also scored and compared with p53 expression. RESULTS: There was no significant difference between inflammatory scores in both groups. In the synovial tissues of rheumatoid arthritis patients, p53 positive cells were detected in 3 out of 9 samples(33%) and p53 expressions were restricted to inflammatory mononuclear cells, but synovial lining cells, subsynovial fibroblast-like cells and vascular endothelial cells were p53 negative. p53 expressions in osteoarthritis synovial tissues as control were observed in 2 out of 5 samples(40%) and the amount and pattern of p53 positive cells were comparable to those seen in rheumatoid arthritis synovial tissues. There was no demonstrable correlation between the synovial tissues of both groups with respect to inflammation scores and expression of p53 protein. CONCLUSION: Our findings suggest that altered p53 expression may not play a significant role in the manifestation of rheumatoid arthritis synovium. However these data need to be strengthened by increasing the number of samples and molecular biology approaches.
Arthritis, Rheumatoid/metabolism* ; Arthritis, Rheumatoid/genetics ; Comparative Study ; Gene Expression ; Genes, p53 ; Human ; Immunohistochemistry ; Osteoarthritis/metabolism ; Osteoarthritis/genetics ; Protein p53/metabolism* ; Protein p53/genetics ; Synovial Membrane/metabolism

OBJECTIVETo explore the expression of interleukin 18 (IL-18) and prostaglandin E2 (PGE2) and their relationship in the synoviocytes of patients with osteoarthritis (OA).

METHODSThe synovial tissues were obtained from 30 OA patients to isolate the synoviocytes for primary culture. The concentrations of IL-18 and PGE2 in the supernatants of synoviocyte culture were measured by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe concentration of IL-18 averaged 51.559-/+27.614 pg/ml and PGE2 327.036-/+333.561 pg/ml in the supernatant of the synoviocytes. A significant positive correlation was noted between their expressions (r=0.863, P<0.01).

CONCLUSIONIL-18 may induce the production of PGE2, and their interactions they may play an important role in the pathogenesis of OA.

Adult ; Aged ; Aged, 80 and over ; Cells, Cultured ; Dinoprostone ; metabolism ; Female ; Humans ; Interleukin-18 ; metabolism ; Male ; Middle Aged ; Osteoarthritis ; metabolism ; pathology ; Synovial Membrane ; metabolism ; pathology