中性粒细胞增强类风湿关节炎滑膜细胞产生基质金属蛋白酶及其细胞侵袭力

目的：研究中性粒细胞表面CD147分子对类风湿关节炎（RA）成纤维样滑膜细胞（FLS）产生基质金属蛋白酶（MMP）及细胞侵袭力的作用。方法：取自关节镜或骨科手术治疗的RA、骨关节炎（OA）患者的滑膜组织，体外分离培养FLS，通过流式细胞术（FCM）检测细胞表面分子及形态学方法鉴定。采用全反式维甲酸（ATRA）诱导HL-60细胞（人早幼粒白血病细胞株）诱导中性粒细胞分化，并用硝基蓝四氮唑（NBT）还原反应测定其分化程度。进而以FCM测定分化前后的HL-60细胞及FLS表面CD147的表达，并通过明胶酶谱和重组基质侵袭实验检测分化前后HL-60细胞对FLS MMP表达和细胞侵袭能力的作用以及CD147拮抗肽（AP-9）对这种MMP表达和细胞侵袭能力的抑制作用。结果：成功建立了FLS分离培养和HL-60细胞分化的实验体系。培养的FLS具有典型成纤维细胞的形态，均一的高度表达CD90（〉98％）而不表达CD14。分化后的HL-60细胞CD147表达上调，RAFLS细胞CD147表达水平低于分化前及分化后HL-60细胞（P〈0．05）。明胶酶谱试验显示：①与OAFLS相比，单培养的RAFLS表达较高水平的酶原及活化形式的MMP-2（P〈0．05）；②分化前／后的HL-60细胞分别与FLS共培养，酶原及活化形式的MMP-2和MMP-9的表达水平较这些细胞单独培养均显著升高（P〈0．05），且分化后的HL-60细胞与FLS共培养酶原及活化形式MMP-2表达水平较分化前HL-60细胞与FLS共培养明显升高（P〈0．05）；③AP-9显著抑制两共培养组中MMP-2和MMP-9的上调作用（P〈0．05）。侵袭实验显示：RAFLS侵袭能力高于OA FLS（P〈0．05）；分化前后的HL-60均可增加RAFLS的侵袭能力（P〈0．05），且分化后强于分化前（P〈0．05）；AP-9可抑制这一促进作用，降低FLS的侵袭能力（P〈0．05）。结论：中性粒细胞可能通过CD147促进RA患者滑膜FLS MMP-2、MMP-9的表达和活化，增强FLS的侵袭能力，这种促进作用可被CD147拈抗肽所抑制，将为RA关节软骨和骨损伤机制和治疗的进一步研究提供了重要的线索。     

Identification of a population of large granular lymphocytes obtained from the rheumatoid joint coexpressing the CD3 and CD16 antigens.

In this study, we phenotypically characterized large granular lymphocytes (LGL) among the synovial fluid mononuclear cells obtained from rheumatoid arthritis (RA) patients. Cytochemical and flow cytometric studies revealed an increased percentage of LGL in the synovial fluid mononuclear cells obtained from patients with RA compared to those without RA. Flow cytometric analysis revealed an expanded population of cells expressing a rare phenotype: CD3 +/CD16+. While these cells are seen in very low percentages in normal individuals (less than 2%) and have been reported in T cell lymphoproliferative disorders. In 20/30 RA patients studied, these cells constituted from 20 to 80% of the total synovial fluid mononuclear cells. Furthermore, studies of synovial fluids with greater than 20% CD3 +/CD16+ cells failed to show significant cytolytic activity even after incubation with recombinant interleukin-2, while fluids with less than 20% CD3 +/CD16+ cells possessed normal cytolytic activity. Studies of matched blood and fluid in eight patients with RA demonstrated a significantly increased percentage of CD3 +/CD16+ cells in synovial fluid compared to peripheral blood. Modulation and adsorption studies did not provide evidence that the CD16 antigen present on these CD3 cells was due to passive adsorption of soluble CD16 antigens. Thus, while the relevance of these cells in the pathogenesis of RA is not clear, this report identifies CD3 +/CD16+ cells in a disease state other than T cell lymphoproliferative disorders.     

Detection of T-Lymphocyte Subpopulations in the Peripheral Blood and the Synovium of Patients with Rheumatoid Arthritis and Juvenile Rheumatoid Arthritis Using Monoclonal Antibodies

Monoclonal antibodies with specificities for various human T-cell antigens were used in direct immunofluorescence to quantify the proportions of T lymphocytes with suppressor/cytotoxic-cell markers and with helper/inducer-cell markers and of T lymphocytes with HLA-DR antigens. Normal percentages of lymphocytes with suppressor/cytotoxic-cell markers were detected in the peripheral blood synovial fluid and synovial tissue lymphocytes from patients with juvenile rheumatoid arthritis (JRA) and rheumatoid arthritis (RA), respectively. Normal percentages of T lymphocytes with helper/inducer-cell markers were seen in the peripheral blood of RA and JRA patients and in the synovial tissues of RA patients. Slightly decreased percentages of cells with the helper/inducer-cell marker were detected in the synovial fluids of JRA patients. The proportions of HLA-DR-positive T lymphocytes were highly increased in the synovial fluid and synovial tissue, whereas the numbers of these cells in the peripheral blood were normal. No significant differences in T gamma cells were detected between peripheral blood, synovial fluid and synovial tissue of JRA patients or between peripheral blood and synovial tissue of RA patients.     

Excessive production of interleukin 6/B cell stimulatory factor-2 in rheumatoid arthritis

High levels of interleukin 6 (IL 6/B cell stimulatory factor-2) were detected in synovial fluids from the joints of patients with active rheumatoid arthritis (RA). The cells found in freshly isolated synovial fluid constitutively expressed IL 6 mRNA. The synovial tissues obtained by joint biopsy were also found to produce IL 6 in vitro. Immunohistochemical analysis demonstrated that CD2+ T cells as well as CD20+ blastoid B cells in the synovial tissues produce IL 6. The data indicate that IL 6 is generated constitutively in RA and its overproduction may explain the local as well as the generalized symptoms of RA, since IL 6 can function as B cell growth and differentiation factor as well as hepatocyte-stimulating factor.     

类风湿关节炎患者外周血和滑液中CD4~+CD25~+调节性T细胞的水平变化

了解具有抑制功能的CD4+CD25+调节性T细胞(Treg)在类风湿关节炎(RA)中的水平变化。分离32例RA患者及35例正常对照者外周血和15例RA关节滑液中的单个核细胞,用荧光抗体标记细胞膜表面CD4、CD25分子和细胞内Foxp3转录因子,进行流式细胞分析,同时用RT-PCR方法测定单个核细胞中Foxp3 mRNA水平。实验发现RA外周血中CD4+CD25hT细胞比例(1.90±1.68)与健康人(1.81±1.79)无明显差异,而RA关节滑液中CD4+CD25+和CD4+CD25hT细胞含量却明显增高(14.98±12.52,8.94±9.67,P<0.01)。RA患者外周血单个核细胞中Foxp3+/CD4+T细胞比值(2.35±2.06)较正常人(7.25±3.98)明显降低(P<0.01),RA外周血中Foxp3 mRNA含量较正常人Treg减少,而RA关节液中Foxp3 mRNA含量较RA外周血更为低下(P<0.01)。RA患者存在CD4+CD25+Treg的异常改变,其外周血和关节液中具有抑制作用的Treg含量明显降低提示RA患者Treg数量减少及抑制功能下降可能是RA自身免疫反应亢强不能控制的原因之一。RA关节液中CD4+CD25hT细胞增高考虑与RA炎症反应造成T细胞过度活化有关。     

Analysis of T cell receptors in rheumatoid arthritis: the increased expression of HLA-DR antigen on circulating gamma delta+ T cells is correlated with disease activity.

The phenotypic characteristics of peripheral blood T cells, isolated from 37 rheumatoid arthritis (RA) patients and 17 healthy controls were determined with special emphasis on gamma delta+ T cells and CD4-CD8- alpha beta+ T cells. Two- and three-colour automated flow cytometry analyses were performed using a panel of MoAbs directed against differentiation antigens and T cell receptor molecules. The results demonstrated: (i) no significant difference between the percentages of CD4-CD8- alpha beta+ T cells in patients and controls; (ii) a significant decrease of the gamma delta+ T cell level in the peripheral blood of RA patients relative to controls; (iii) phenotypic abnormalities of circulating gamma delta+ T cells in RA patients suggestive of an activation status in vivo. These abnormalities included a significant reduction in the density of the T cell differentiation antigen CD3 and an increase in the expression of HLA-DR antigen. The level of circulating HLA-DR+/gamma delta+ T cells was significantly higher in patients with active disease. HLA-DR+/gamma delta+ T cells were also present in the synovial fluid obtained from three patients with an active disease. In addition, preliminary experiments showed that the activated gamma delta+ T cells were predominantly V delta 1. Taken together, these data support the involvement of gamma delta+ T cells in the pathogenesis of RA.     

Expression of CD40 and CD40 ligand among cell populations within rheumatoid synovial compartment

Augmented and prolonged expression of CD40 ligand (CD40L) was recently reported in lymphoid cells from human lupus patients, suggesting that CD40/CD40L pathway was involved in the pathogenesis of systemic autoimmune diseases. This study was thus designed to study the expression of CD40 and CD40L among cell populations within inflammatory joints of patients with rheumatoid arthritis (RA). The result showed that most B cells and monocytes in synovial fluids (SF) expressed CD40. Cultured synovial fibroblasts also stained positive for CD40. Regarding CD40L, we found that T cells as well as B cells could express CD40L. Compared with normal controls, RA patients had higher levels of CD40L+ T cells (8.71 +/- 17.69% vs 1.74 +/- 2.30%, P > 0.05) and CD40L+ B cells (7.71 +/- 7.64% vs 1.12 +/- 1.59% P < 0.05). After in vitro stimulation, T cells from RA patients had higher and longer CD40L expression than T cells from normal peripheral blood. For investigating the effect of CD40 expressed on synovial fibroblasts on TNF-alpha production in joint compartment, we used anti-CD40 antibody to bind CD40 on fibroblasts for one hour and then co-cultured with synovial fluid mononuclear cells. We found that the levels of TNF-alpha decreased in the presence of anti-CD40 antibody. We concluded that there was an intrinsic hyperexpression of CD40L on lymphoid cells within rheumatoid joints, and synovial fibroblasts could contribute to articular inflammation through surface CD40 molecule.     

A high proportion of the V delta 1+ synovial fluid gamma delta T cells in juvenile rheumatoid arthritis patients express the very early activation marker CD69, but carry the high molecular weight isoform of the leucocyte common antigen (CD45RA).

We have previously shown that gamma delta T cells in the synovial compartment of patients with juvenile rheumatoid arthritis (JRA) express activation antigens (CD69 and HLA-DR) and that they are predominantly of the V delta 1 subset. In this study we have analysed the expression of activation antigens (CD69 and HLA-DR) and different isoforms of the leucocyte common antigen (CD45RO and CD45RA) on the V delta 1 and the V delta 2 subsets of gamma delta T cells in paired samples of synovial fluid and peripheral blood of nine patients with JRA, and in the peripheral blood of five children with idiopathic scoliosis. In the synovial fluid of children with JRA, there were significantly more V delta 1+CD69+ and V delta 2+CD69+ cells compared with the peripheral blood of the same patients. In contrast, however, in the synovial fluid the V delta 1 and the V delta 2 subsets differed with respect to the expression of the two isoforms of the leucocyte common antigen. The majority of the V delta 1+ cells expressed the high molecular weight isoform (CD45RA+) while most of the V delta 2+ cells carried the low molecular weight variant (CD45RO+) of this molecule.     

Synovial fluid macrophages are capable of osteoclast formation and resorption

To determine whether synovial fluid (SF) macrophages isolated from the SF of osteoarthritis (OA), rheumatoid arthritis (RA) and pyrophosphate arthropathy (PPA) joints are capable of osteoclast formation, and to investigate the cellular and humoral factors required for this to occur, SF macrophages (CD14+) were isolated from the knee joint SF from patients with OA, RA and PPA and cultured for up to 14 days with macrophage-colony stimulating factor (M-CSF) and soluble receptor activator for nuclear factor-kappaB ligand (RANKL) or tumour-necrosis factor-alpha (TNFalpha) and interleukin-1alpha (IL-1alpha). Osteoclast differentiation was assessed by expression of tartrate-resistant acid phosphatase (TRAP) and vitronectin receptor (VNR), F-actin ring formation and lacunar resorption. Osteoclast formation and lacunar resorption was seen in RANKL-treated cultures of SF macrophages isolated from OA, RA and PPA joints with the largest amount of resorption noted in RA and PPA SF macrophage cultures. In TNFalpha/IL-1alpha-treated RA and PPA SF macrophage cultures, osteoclasts capable of lacunar resorption were also formed. Lacunar resorption was more extensive in RANKL than TNFalpha/IL-1alpha-treated cultures. These findings indicate that SF macrophages are capable of differentiating into mature osteoclasts capable of lacunar resorption. M-CSF in combination with RANKL or TNFalpha/IL-1alpha was required for osteoclast formation. As inflammatory synovial fluids contain an increase in the number of macrophages and an increase in the amounts of RANKL, TNFalpha and IL-1alpha, these findings suggest that one means whereby bone erosions may form in rheumatoid or crystal arthritis is by differentiation of synovial fluid macrophages into osteoclasts.     

Suppressive influences of methotrexate on the generation of CD14(+) monocyte-lineage cells from bone marrow of patients with rheumatoid arthritis

An adequate supply of peripheral blood monocytes, granulocytes, and platelets is necessary for an optimal inflammatory process. We have previously demonstrated that the generation of CD14(+) monocyte lineage cells from the bone marrow is accelerated in patients with rheumatoid arthritis (RA). The current studies examined the influences of methotrexate (MTX), a potent disease modifying antirheumatic drug (DMARD), on the capacity of bone marrow progenitor cells to generate CD14(+) cells in patients with RA, in order to delineate its mechanism of action. CD14(-) cells purified from bone marrow specimens of 14 patients with active RA were cultured in the presence or the absence of pharmacologically attainable concentrations of MTX (0.2 microM). After incubation for 14 days, the cells were analyzed by flow cytometry for expression of CD14 and HLA-DR. The generation of CD14(+) cells from RA bone marrow CD14(-) progenitor cells was significantly suppressed by MTX. However, the expression of HLA-DR on bone marrow-derived CD14(+) cells was not significantly influenced by MTX. There was no significant difference in the effect of MTX on the generation of CD14(+) cells between patients with prednisolone and those without prednisolone. The production of IL-12 in bone marrow cell cultures was not inhibited, but was rather enhanced, by MTX, suggesting that the suppression of the generation of CD14(+) cells might not be due to the inhibition of cytokine production. The results are consistent with the hypothesis that one of the effects of DMARDs may involve the interference with monocyte differentiation in the bone marrow. Moreover, the data suggest that the generation of CD14(+) cells and the expression of HLA-DR on such marrow-derived CD14(+) cells are regulated by different mechanisms.     

Expression of extra trinucleotide in CD44 variant of rheumatoid arthritis patients allows generation of disease-specific monoclonal antibody

Selective targeting of cells engaged in pathological activities is a major challenge for medical research. We generated monoclonal antibodies (mAbs) that exclusively bind, at concentrations ranging from 2 to 100 microg/ml, to a modified CD44 variant (designated CD44vRA) expressed on synovial fluid cells from joints of rheumatoid arthritis (RA) patients. These mAbs cross-reacted with keratinocytes expressing wild type CD44vRA (CD44v3-v10) only at a relatively high concentration (200 microg/ml). Sequence analysis of CD44vRA cDNA revealed, in 33 out of 43 RA and psoriatic arthritis patients, an extra intron-derived trinucleotide, CAG, which allows translation of an extra alanine. This insertion imposes a configurational change on the cell surface CD44 of RA synovial fluid cells, creating an immunogenic epitope and potentiating the ability to produce disease-specific antibodies. Indeed, the anti-CD44vRA mAbs (designated F8:33) were able to induce apoptosis in synovial fluid cells from RA patients, but not in peripheral blood leukocytes from the same patients, in keratinocytes from normal donors or in synovial fluid cells from osteoarthritis patients. Furthermore, injection of anti-CD44vRA mAbs reduced joint inflammation in DBA/1 mice with collagen-induced arthritis. These findings show that anti-CD44vRA mAbs are both bioactive and RA-specific.     

Studies on CD11a and CD18 molecules with two new monoclonal antibodies: differential myelomonocytic antigen expression of PMA treated HL60 and U937 cell lines

Biochemical and functional aspects as well as features of cellular distribution of the differentiation groups CD11a and CD18 were studied in the course of a detailed characterization of two new monoclonal antibodies which recognize the alpha (GRS3) and beta (GRF1) chains of the LFA-1 antigen. Both MAbs inhibited homotypic aggregation of an EBV cell line. In contrast, only GRF1 (anti-beta chain) was able to inhibit granulocyte aggregation as well. Different myeloid-monocyte antigen modulation was noted in PMA induced macrophage differentiation of the U937 and HL60 cell lines. PMA treated HL60 cells showed increased expression of alpha M (CD11b) and alpha X (CD11c) antigens but no change in HLA-DR or CD14 antigen expression. No variation in the expression of LFA complex antigen (CD11a, b and c, or CD18) was observed on U937 cells, which on the other hand presented de novo expression of HLA-DR and CD14 antigens.     

Synovial fluid cells in juvenile arthritis: evidence of selective T cell migration to inflamed tissue.

The perpetuation of chronic synovitis in juvenile arthritis (JA) is a complex interaction of local and systemic regulatory mechanism. We examined the cell surface phenotype of synovial fluid cells and peripheral blood lymphocytes from 15 patients with JA to better understand the mechanism of local inflammation. Synovial fluid and peripheral blood mononuclear cells were analysed for cell surface expression of CD2, CD3, CD4, CD8, CD19, CD25, CD29, CD45R and Ia using flow cytometry. We found a very low percentage of B cells with a concomitant increase of T cells in synovial fluid as compared with peripheral blood. A large percentage of the synovial fluid T cells were HLA-DR+, or activated T cells, and there was a relative decrease in CD4+ cells in synovial fluid as compared with peripheral blood. There was only a minimal increase in CD25+ synovial fluid cells. The synovial fluid CD4+ cells were mainly of the CD2high, CD29+, CD45RO phenotype. This CD4 phenotype found on synovial fluid cells from patients with JA and in particular the CD29 cell surface marker, which recognizes a common beta-chain of adhesion molecules, is associated with binding to extracellular matrix proteins and is also associated with 'primed' T cells. Our results demonstrated the presence of T cells which either selectively migrate to synovium and synovial fluid or are activated in situ in the joint.     

Generation of interleukin-2-dependent T cell lines from synovial fluids in rheumatoid arthritis.

Synovial fluids from rheumatoid arthritis (RA) patients were found to contain activated T lymphocytes that could be maintained as continuous T cell lines (CTCL) in the presence of the T cell growth factor, interleukin (IL)-2. The CTCL predominantly expressed the OKT8 phenotype and were Ia antigen positive. IL-2-dependent RA CTCL could be maintained in an active dividing state by the presence of RA synovial fluids, whereas IL-2-dependent CTCL from mitogen stimulated PBL failed to respond to the fluids, which were shown to contain IL-2. This suggested that RA CTCL exhibit unique properties not possessed by normal PBL CTCL. The CTCL generated from activated synovial T lymphocyte populations in RA may be used to assess the functions of these cells and their responses to regulatory factors.     

1,25-Dihydroxyvitamin D3 and phorbol esters (TPA) may induce select in vitro differentiation pathways in the HL60 promyelocytic cell line

Monocytic features can be induced in the myeloid cell line HL60 in order to provide a suitable in vitro model for the investigation of in vitro activity in mononuclear phagocytes. 1,25-Dihydroxyvitamin D3 (calcitriol) induced the HL60 cell line to express the monocytic differentiation antigen Leu M3 in about 30-50% of the cells along with an increase (up to 20%) in the expression of HLA-DR but not HLA-DQ class II antigen. Functional investigation showed that calcitriol-treated cells formed rosettes with sheep erythrocytes coated with an anti-sheep erythrocyte-specific IgG2a mouse MoAb and readily ingested them. In addition, these same sensitized erythrocytes were lysed in an 18-hr antibody-dependent cellular cytotoxicity (ADCC) assay. All together these data indicate the presence of functionally active Fc-IgG receptors (FcR). Sorting experiments demonstrated that only Leu M3+ HLA-DR+ cells contained the effector cell population; such was also the case for blood monocytes. This phenotypic profile was, however, not predictive per se of FcR presence and function, as 12,O-tetradecanoylphorbol-13-acetate (TPA)-induced HL60 cells neither formed rosettes nor phagocytosed nor exhibited ADCC activity, although they express Leu M3 and HLA-DR (as well as HLA-DQ) antigens. These results suggest that calcitriol and TPA cause the differentiation of HL60 cells along distinct pathways. On the other hand, different subpopulations with given predetermined differentiation capabilities may coexist in HL60 cell line. This hypothesis gains support by the observation that when TPA and calcitriol were added together to the undifferentiated cells, the resulting phenotypic pattern was representative of the different activities of both of the inducers as they were used separately.     

人关节液促进骨髓间充质干细胞的定向分化

背景：目前的众多研究都是通过体外加入生长因子诱导骨髓间充质干细胞分化,但该方法细胞因子用量大,成本高,随着培养次数的增加,其成软骨潜能明显降低。 目的：混合培养人关节液与人骨髓间充质干细胞,观察共培养系统对人骨髓间充质干细胞定向诱导分化的影响。 方法：采用全骨髓、贴壁培养法培养和分离人骨髓间充质干细胞,无菌操作下抽取健康志愿者的膝关节液,将其与P3代细胞混合用于实验,分为3组,关节液+完全培养基,关节液+人骨髓间充质干细胞+完全培养基,人骨髓间充质干细胞+完全培养基。每天在倒置显微镜下观察细胞形态变化和生长情况,分别于诱导后第7,14,21天行甲苯胺蓝染色检测和Ⅱ型胶原免疫组化染色检测。 结果与结论：人关节液与人骨髓间充质干细胞混合培养后,细胞增殖速度减慢,由长梭形变为多角形、椭圆形,细胞外基质呈甲苯胺蓝异染性阳性,Ⅱ型胶原免疫组化染色阳性。说明关节液对人骨髓间充质干细胞向软骨细胞分化有正性促进作用,关节液中可能含有促人骨髓间充质干细胞向软骨细胞方向分化的物质。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

Polyreactivity of human IgG Fc-binding phage antibodies constructed from synovial fluid CD38+ B cells of patients with rheumatoid arthritis

Recent data indicate that rheumatoid factors (RFs) that occur in patients with rheumatoid arthritis (RA) are derived from Ig-producing terminally differentiated CD20-, CD38+ plasma cells present in synovial fluids (SFs). Phage antibody display libraries were constructed using CD38+ plasma cells isolated from SFs of two RF-seropositive RA patients. The libraries were enriched for phage antibodies (Phabs) binding to human IgG (HuIgG) Fc fragments and the sequences of their V genes were analysed. These data provided further evidence for an Ag-driven immune response in patients with RA, including expansion of clonally related B cells, selection and isotype switching, all hallmarks of a germinal center reaction. In the present study, the functional characteristics of these HuIgG Fc-binding monoclonal (mo) Phabs were further analysed in order to provide more insight into the specificity of HuIgG Fc-binding Phabs. Remarkably, all HuIgG Fc-binding moPhabs tested (n=48; derived from four different libraries) displayed polyreactivity. Structural analysis of the CDR3 regions revealed characteristic features of polyreactive Igs. Most H chain CDR3 regions harboured tryptophan/tyrosine-rich parts and approximately 60% of the L chain CDR3 regions of both RA patients displayed an identical stretch of amino acids (W/Y-D-S-S). Supportive for a dominant role of VH in specificity, exchange of VL regions with a single VH region yielded moPhabs with similar specificities. All together, the data suggest the presence of an Ag-driven process in the joints of patients with RA, including somatic mutation and clonal selection entailing isotype switching, resulting in the differentiation of B cells into polyreactive RF-secreting plasma cells.     

Soluble Forms of P-Selectin and Intercellular Adhesion Molecule-3 in Synovial Fluids

A number of adhesion molecules have been identified in synovial tissues of patients with rheumatoid arthritis (RA). Some of them are upregulated and may play an important role in the inflammatory processes of the diseased joint. In addition to synovial tissue cell surface expression, synovial fluids contain soluble forms of many adhesion molecules, such as intercellular adhesion molecule-1, E-selectin (sE-selectin), and L-selectin. In this study, we investigated the expression of soluble P-selectin (sP-selectin) and intercellular adhesion molecule-3 (sICAM-3) in synovial fluids from patients with RA, osteoarthritis (OA), and other forms of arthritis. sP-selectin and sICAM-3 levels in RA synovial fluids were significantly increased compared to those in OA. The levels of sP-selectin in synovial fluids correlated with sICAM-3 and sE-selectin in synovial fluids. The levels of sICAM-3 in synovial fluids correlated with synovial fluid leukocyte counts and erythrocyte sedimentation rate.In vitro,synovial fluid mononuclear cells produced sICAM-3 spontaneously. Elevated levels of sP-selectin and sICAM-3 in RA synovial fluids compared to OA may indicate inflammatory interactions between endothelial cells, leukocytes, and other synovial cells in the diseased joint.     

二十碳五烯酸联合视黄酸对HL-60细胞增殖与分化功能的影响

目的研究二十碳五烯酸 ( eicosapentaenoic acid,EPA)及其联合视黄酸 ( retinoic acid,RA)对白血病细胞增殖与分化功能的影响。方法采用 MTT法测定细胞增殖功能 ,NBT还原实验鉴定细胞分化 ,高效液相色谱法分析细胞内 RA代谢。结果与对照组相比 ,EPA组细胞增殖抑制率为 2 4 .3 8% ,RA组为 3 5 .74 % ,EPA+ RA组为 4 2 .75 % ;NBT还原实验 ,EPA+ RA组 OD值约为对照组的 5 .9倍 ,而 RA组为 2 .6倍 ,EPA组为 1.3倍 ;分析 HL- 60细胞及其培养介质中 RA含量 ,发现 EPA+ RA组高于 RA组。结论 EPA可抑制 HL- 60细胞增殖和诱导其分化 ,与 RA联合应用能明显增强 HL- 60细胞的增殖抑制及诱导分化效应 ,这种联合效应可能与 EPA减缓 HL - 60细胞内 RA的代谢相关。