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Levels of Soluble CD27 in Sera and Synovial Fluid and Its Expression On Clinical and Experimental Rheumatology 2002; 20: 863-866. PEDIATRIC RHEUMATOLOGY Levels of soluble CD27 in ABSTRACT man inflammatory arthritides, includ- sera and synovial fluid O b j e c t ive. CD27 is a member of ing juvenile idiopathic arthritides (JIA) tumour necrosis factor receptor family. (1-3). and its expression on Its ex p ression is pre d o m i n a n t ly con - CD27 is a member of the tumour ne- memory T cells in fined to mat u re ly m p h o cytes and is crosis factor receptor (TNFR) family. patients with juvenile strongly enhanced after cell activation. Its ex p ression is pre d o m i n a n t ly con- idiopathic arthritides Shedding of the CD27 from the surface fined to mat u re ly m p h o cytes and is of activated cells is re l ated to their strongly enhanced after cell activation effector phase.The aim of the present (4). This molecule has been shown to 1 2 M. Gattorno , I. Prigione , study was to evaluate the levels of solu - stimulate T cell proliferation and B cell S. Vignola3, F. Falcini4, ble CD27 in sera and synovial fluids, differentiation through the interaction S. Chiesa2, F. Morandi2, together with its expression on circulat - with its specific ligand, namely CD70. P. Picco1, A. Buoncompagni1, ing and synovial fluid (SF) memory T Pe rsistent antigenic stimu l ation in- cells, in children with JIA. duces the proteolytic cleavage of the 1 2 A. Martini , V. Pistoia Methods. Sera from 40 patients with soluble 32 kDa form of CD27 from the a c t ive JIA we re studied for solubl e cell surface (4). According to previous 1 Second Department of Pediatrics CD27. Paired SF samples were avail - studies such process correlates with the (Rheumatology Unit), 2Laboratory of Oncology and 3Third Department of Pedi- able in 20 patients. Sera from 12 age- degree of T cell activation, both at the atrics (Gastroenterology Unit); “G. Gasli- matched patients affected with various systemic and the local level. In fact, ni” Institute for Children, Genoa; 4Clinics acute infectious diseases and 12 age- increased levels of soluble CD27 have of Pediatrics, Florence, Italy. matched healthy subjects were used as been found both in sera of renal trans- Marco Gattorno, MD; Ignazia Prigione, controls. In 8 JIA patients freshly iso - plant recipients (5) and in cerebrospi- PhD; Silvia Vignola, MD; Fernanda Falci- lated circulating and SF lymphocytes nal fluid of patients affected with mul- ni, Associate Professor of Pediatrics; Sab- were stained for CD27 in CD4+CD45 tiple sclerosis (6). rina Chiesa, PhD; Fabio Morandi; PhD; Paolo Picco, MD; Antonella Buoncompag- RO+ T cell subpopulation and ana - According to the model proposed by ni, MD; Alberto Martini, Professor of Pedi- lyzed by cytometry. Van Lier et al., naïve T cells constitu- atrics; Vito Pistoia, MD. R e s u l t s . S o l u ble CD27 serum leve l s tively express CD27, which is up-regu- Please address correspondence and reprint we re signifi c a n t ly higher in pat i e n t s l ated fo l l owing antigen pre s e n t at i o n requests to: Marco Gattorno, MD, 2nd with polyarticular JIA and acute sys - and T cell activation. Interactions be- Department of Pediatrics, “G. Gaslini” temic infectious diseases than in tween the T cell co-stimulatory mole- Scientific Institute for Children and Uni- patients with active oligoarticular or cule CD28 and its ligands (CD80/CD versity of Genoa, Largo G. Gaslini 5, h e a l t hy controls. Both polya rt i c u l a r 86) on antigen presenting cells (APC) 16147, Genoa, Italy. and oligoarticular JIA patients showed is followed by the release of pro-in- E-mail: marcogattorno@ospedale-gasli- ni.ge.it increased levels of soluble CD27 in SF flammatory cytokine from APC, which when compared with paired seru m eventually results to the expression of Received on February 28, 2002; accepted in revised form June 4, 2002. samples (p = 0.01). In all the patients CD70 on activated T cells. The CD27- tested a significant enri chment of CD70 interaction will further support © Copyright CLINICAL AND EXPERIMENTAL RHEUMATOLOGY 2002. CD27– T cells was seen in the SF clonal expansion of antigen specific T (median 39.5%, range 18-56%) when cells, the majority of which will subse- Key words: Juvenile idiopathic compared to paired CD4+CD45RO+ quently differentiate into effector cells, arthritides, memory T cells, activation, pe ri p h e r al lym p h o c ytes (median 19.5%, shedding the CD27 from their surface CD27. range 5-43%; p = 0.01). in fluid phase. C o n cl u s i o n s . A clear enri chment of Conversely, a subset of antigen-specific CD4+ memory SF T cells with a CD27- T cells that does not differentiate into phenotype is observed when compared effector cells circulate in the blood as to correspondent circulating T lympho - CD27+ “resting” memory T cells, rea- cytes. This issue is conceivably related dy to undergo to re-activation upon sec- to re - a c t ivation and re c ruitment of o n d a ry ex p o s u re to antigen (“early ” memory T cells to the site of inflamma - m e m o ry) and subsequent diffe re n t i a- tion, and to the subsequent expansion tion into “ e ffe c t o r ” C D 2 7- m e m o ry of a subpopulation of “effector” memo - cells (7). ry T cells. The concept the CD4+CD45RO+CD27- cell subset associates with the effector Introduction phase of memory T cells differentiation T lymphocytes are thought to play a is supported by the following observa- pivotal role in the initiation and perpet- tions: i) it is a potent inducer of B cell uation of chronic synovitis in the hu- differentiation (and Ig production) (8); 863 PEDIATRIC RHEUMATOLOGY CD27 in juvenile idiopathic arthritides / M. Gattorno et al. ii) it is enriched for cells producing IL- Pa i red synovial fluid (SF) samples weeks before the examination, together 4 or IFN-g ( 9 , 10); iii) it ex p re s s e s were collected after therapeutic arthro- with clinical or laboratory (i.e. elevated organ-specific homing receptors, such centesis in 20 patients (7 with poly- ESR or CRP) signs of inflammation at as CLA and the a Eb7 integrin (11, 12). a rticular and 13 with oligo a rt i c u l a r the moment of the study, were consid- Soluble CD27 has been found to be sig- course). ered as criteria of exclusion. n i fi c a n t ly increased in the synov i a l A number of clinical (no. of active S o l u ble CD27 in sera and SF wa s fluid of patients affected with rheuma- joints, no. of joints with limited range determined using a commercial ELISA toid art h ritis (RA) in comparison to of motion and physician global assess- kit (CLB,The Netherlands), according patients with osteoarthritis (13). More- ment of overall disease activity) and to the manufacturer’s instructions. over, CD27 has been shown to be ex- laboratory parameters (erythrocyte sed- Expression of CD27 on peripheral and pressed in different proportion and dis- imentation rate, ESR; C-reactive pro- SF memory T cells. Peripheral blood tribution, particularly in the subset of tein, CRP; white blood cell, WBC and (PB) and synovial fluid (SF) mononu- me m o r y T cells (CD4+CD45RO+) both platelet, PLT, counts; hemoglobin ser- clear cells were isolated from heparin- in the synovial fluid and in the inflam- um concentration, Hb) of disease activ- ized blood and synovial samples from 8 ed synovial tissue from RA pat i e n t s ity we re re c o rd e d, t ogether with the JIA patients (2 poly and 6 oligo) by (13, 14). ongoing treatment, at the time of the Ficoll (Biochrom KG-Germany) densi- Aim of the present study was to evalu- study (Table I). ty gradient centrifugation. PB mononu- ate the levels of soluble CD27 in sera Disease activity was defined by the pre- clear cells were also isolated from two and synovial fluids, together with its sence of active arthritis (swelling or, if age - m at ched healthy controls. Cells expression on circulating and synovial swelling was ab s e n t , l i m i t ation of were washed and a three-color staining fluid memory T cells, f rom ch i l d re n motion with tenderness) at least in one was performed with CD45RO-tricolor affected with JIA. joint at the moment of clinical exami- mAb (Caltag - B u rl i n ga m e - C a ) , C D 4 - nation and a physician global estimate FITC or -PE mAb (BD Biosciences- Patients and methods of disease activity (measured on a 0-10 Mountain View-Ca) and CD27-PE or - Soluble CD27 serum and SF cm visual analog scale) higher than 1 FITC mAb (PharMingen, San Diego, concentrations (16,17). CA). Cells were incubated with saturat- Sera from 40 consecutive patients af- Se r a from 12 previ o u s l y healthy age- ing amounts of mAbs for 30’ at 4°C, fected with active JIA according to mat c hed patients affected with vari o u s then cells we re washed in PBS Durban criteria were studied between acute feb rile infectious conditions (4 (Biowhittaker-Walkersville-USA) with (15).
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