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Medical Inflammation Research, Lund University, Lund, Sweden
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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T cells, which target peripheral joints, transfer PIA. The pristane-primed T cells are of oligo or polyclonal origin as determined by their arthritogenicity after stimulation with several mitogenic anti-TCRV and anti-TCRV mAbs. Arthritogenic cells secreted IFN-
and TNF- (but not IL-4) when stimulated with Con A in vitro, and pretreatments of recipient rats with either anti-IFN- or a recombinant TNF- receptor before transfer ameliorated arthritis development. Most importantly, we show that these T cells are MHC class II restricted, because treatment with Abs against either DQ or DR molecules ameliorates arthritis development. The MHC class II restriction was confirmed by transferring donor T cells to irradiated recipients that were syngenic, semiallogenic, or allogenic to MHC class II molecules, in which only syngenic and semiallogenic recipients developed arthritis. These data suggest that the in vivo administration of a non-antigenic adjuvant, like pristane, activates CD4+ T cells that are MHC class II restricted and arthritogenic. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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-chain (2). The RA-associated peptide binding pocket in DR molecules may favor presentation of certain self-peptides to autoreactive T cells. Consequently, autoreactive CD4+ T cells, which recognize peptides on MHC class II molecules, may play an important role in RA. Moreover, the vast majority of lymphocytes infiltrating RA synovia are CD4+ T cells (3, 4). The joint-restricted CD4+ T cell clonotypes that persist over time are preferentially found in the CD25+ subpopulation (5). Skewing of the TCRV repertoire (e.g., V3, V14 
V17) has been shown to occur frequently in CD25+ T cells in the synovial fluid, indicating a local activation by a common antigenic stimuli (6). However, it has been difficult to identify clones of arthritogenic T cells or a common Ag in the joint that could explain the development of arthritis, and the pathogenic function of infiltrating T cells into the joint synovia is still uncertain. Several reports show a significant IFN- secretion from synovial fluid mononuclear cells of RA patients, indicating a Th1 response in RA (7, 8).
A major problem in the investigations of the role of T cells in arthritis is the lack of animal models that can efficiently transfer disease with specific T cells. In accordance with this, the most commonly used animal model, collagen-induced arthritis (CIA), cannot be as efficiently transferred compared with the corresponding model for multiple sclerosis, the experimental autoimmune encephalomyelitis (9). Surprisingly, it is well known that adjuvant-induced arthritides can be transferred with T cells (10, 11, 12, 13). There are no evidences that adjuvants, such as pristane, bind MHC class II molecules, and there is no direct link to adaptive autoimmunity. Thus, the specificity or MHC restriction of the arthritogenic T cells is not known. We therefore set out to determine whether pristane-activated T cells are MHC class II restricted and could transfer arthritis. Similar to RA, pristane-induced arthritis (PIA) shows symmetry of inflammation, chronic relapsing disease course, infiltration of T cells, and erosive destruction of cartilagenous peripheral joints (14). Furthermore, the PIA model is characterized genetically, and one of the identified loci contain the MHC region (PIA locus 1 (Pia1)) and another locus contains the TCR locus (PIA locus 5 (Pia5)) (15). In the current study, we show that pristane injection in rats leads to activation of CD4+ T cells that transfer arthritis and occurs in the arthritic synovia. The T cell-induced arthritis was depending on IFN- and TNF-, as shown by Ab-mediated cytokine blockage in vivo. Finally, we show that the arthritogenic T cells are MHC class II restricted, which argues for Ag involvement in this model, although the pristane itself is non-antigenic.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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The DA and DA.1I (originating from Zentralinstitut Fur Versuchstierzucht, Hannover, Germany) rat strains were kept in the animal facility of Medical Inflammation Research in a climate-controlled environment with 12-h light/dark cycles, housed in polystyrene cages containing wood shavings, and fed standard rodent chow and water ad libitum. The congenic DA.1I (D20Rat42 to D20Rat49; n = 10) and the DA.1U (D20Rat47 to AA858870; n = 6) rats produced by speed congenic technique (16) express i and u alleles, respectively, for the rat MHC class I (RT1-A) and class II (RT1-B/D) molecules. Female DA and male DA.1I rats were intercrossed to produce (DAxDA.1I)F1 offspring, and a DA.1Ua/uxDA.1Ua/u intercross was set up to produce littermate offspring that were MHC syngenic (a/a), semiallogenic (a/u), or MHC allogenic (u/u) to DA (a/a). The experiments were approved by a local (Malmö/Lund, Sweden) ethical committee (license M7-01).
PIA induction and evaluation of arthritis
Arthritis was induced at the age of 8–12 wk in age-matched female rats by an intradermal injection at the base of the tail with 500 µl of pristane oil (2,6,10,14-tetramethylpentadecane; Sigma-Aldrich). Arthritis development was monitored in all four limbs using a macroscopic scoring system. Briefly, 1 point was given for each swollen or red toe; 1 point was given for each swollen midfoot, digit, or knuckle; and 5 points were given for a swollen ankle (maximum score per limb and rat was 15 and 60, respectively).
Histological analysis
Twelve days after adoptive transfer of 20 x 106 pristane-primed spleen cells from (DAxDA.1I)F1 donor rats, paws from DA.1I recipient rats were collected and decalcified with EDTA. Cryostat sections were stained with hematoxylin and subjected to immunohistochemical staining with haplotype-specific (RT1-Aa) biotinylated anti-RT1-A mAb (R3/13, clone C3) (17), followed by streptavidin-peroxidase, and visualized using a DAB substrate kit (ChemMate; DakoCytomation).
Isolation of synovial cells
At 14 days after pristane injection and 12 days after T cell transfer, rats were killed, and synovia from swollen talo-crural joints were harvested. This was done by isolating the joint by cutting the bone and removing the skin. The remaining joint capsule was excised. The tissue was homogenized and cultured for 2 h in DMEM supplemented with heat-inactivated FCS (10%), collagenase D (0.60 U/ml; Roche), and DNase 1 (5 U/ml; Sigma-Aldrich). A single-cell suspension was made by passing cells through a 40-µm filter, followed by a 40% Percoll (Amersham Biosciences) density gradient separation to obtain large lymphocytes (18). The cells were then washed and resuspended in PBS-D (Dulbecco’s PBS not containing CaCl2 or MgCl2; Invitrogen Life Technologies), supplemented with 0.5% BSA (Sigma-Aldrich) for Ab staining.
Abs and immunosuppressive reagents
The following anti-rat mAbs used for flow cytometry analysis were purchased from Pharmingen (San Diego, CA) as FITC, PE, or biotinylated conjugates: anti-CD4 (clone OX-35), anti-CD8a (clone OX-8), anti-CD25 (clone OX-39), anti-LCA (leukocyte common Ag) for staining of leukocytes (clone OX-1), anti-CD45RA for staining of B cells (clone OX-33), anti-TCR (clone R73), and anti-RT1Aa,b (clone C3). mAbs used for in vivo blocking experiments were grown from hybridomas purchased from American Type Culture Collection: OX-6 (anti-RT1B) (19), OX-17 (anti-RT1D) (19), DB-1 (anti-IFN-) (20), and Hy2.15 (mouse anti-rat TNP, IgG1 isotype control). Etanercept (Enbrel; Wyeth-Ayerst Pharmaceuticals), a generous gift from Dr. H. Burkhardt (Department of Internal Medicine III and Institute of Clinical Immunology, University of Erlangen-Nuremberg, Erlangen, Germany), was used for in vivo blocking of TNF- (21), using PBS-D as control.
Flow cytometry
Cells to be stained were collected from spleen, peripheral blood, or swollen synovia at different time points after pristane injection or T cell transfer, as indicated in the text. Single cells were prepared and stained with FITC-, PE-, and biotin-conjugated mouse anti-rat mAbs for 20 min at 4°C. Streptavidin-APC was used as a secondary reagent, and propidium iodide was added before acquisition. Lymphocyte expansion in draining lymph nodes and spleen was determined by comparing the number of live cells in each population with the total number of cells in the lymph nodes and spleen, respectively. We acquired a minimum of 104 to 105 cells on a FACScan (BD Biosciences), with gates set to include all viable cells that were further analyzed by CellQuest (BD Biosciences) software.
Cytokine detection
Quantitative measurements of IFN-, TNF-, and IL-4 in cell supernatants were performed using the eBioscience rat IFN- and the rat IL-4 BD OptEIA ELISA Set protocol (BD Pharmingen). The lower limits of detection sensitivity of these assays were 1.6 pg/ml for IL-4 and 15 pg/ml for IFN-. For the rat TNF- ELISA (22), coating anti-rat TNF- (SB/230499/GR), detecting biotin-conjugated anti-rat TNF-, (S54/250499/GR), and TNF- standard (190499) were kindly provided by Dr. S. Poole (Division of Endocrinology, National Institute for Biological Standards and Control, Hertfordshire, U.K.). Briefly, one million spleen cells from five pristane-primed and naive DA rats (day 14 after injection) were stimulated for 48 h at 37°C with or without Con A (3 µg/ml; Sigma-Aldrich) in triplicate, in 200 µl of DMEM supplemented with FCS (5%), HEPES (2.4 mg/ml), 2-ME (3.9 µg/ml), and penicillin-streptamycin (104 IU/ml penicillin, 10 mg/ml streptomycin; Invitrogen Life Technologies). Cytokines were detected by a heterogeneous time-resolved fluorometric assay (Victor/1420 Multilabel counter; Wallac Sweden AB) using Eu3+-conjugated streptavidin (Delfia; PerkinElmer) and Delfia® Enhancement Solution for quantitative determination of Eu3+, reflecting quantities of IFN-, IL-4, and TNF-. All cytokine assays were verified using recombinant proteins as positive controls.
T cell transfer
At 14 days after pristane injection, rats were killed, and inguinal lymph nodes and spleens were removed. Cells were washed and passed through 40-µm filters and reactivated in vitro with Con A (3 µg/ml; Sigma-Aldrich); IL-2 (800 pg/ml in final concentration from an IL-2-transfected X63 cell line from our hybridoma collection); plate-bound anti-rat TCR mAbs anti-TCR (clone R73) (23), anti-TCRV4 (clone G99) (24), anti-TCRV8.5 (clone B73) (25), anti-TCRV10 (clone G101) (25), and anti-TCRV16 (clone HIS42) (26) (5 µg/ml); or an anti-TNP IgG1 isotype control (clone Hy2.15; 5 µg/ml) in DMEM supplemented with FCS (5%), HEPES (2.4 mg/ml), 2-ME (3.9 µg/ml), and penicillin-streptomycin (104 IU/ml penicillin, 10 mg/ml streptamycin; Invitrogen Life Technologies). After 48 h of incubation at 37°C, cells were washed and resuspended in PBS-D. Naive recipient rats were then injected i.v. (through the tail vein) with different numbers of cells as indicated in the text and tables. All cell-sorting experiments were performed according to the Dynabiotech protocol (Dynal Biotech). Anti-CD4 (clone OX-35), anti-CD8a (clone OX-8), and anti-CD25 (clone OX-39) were coated on CELLection Pan Mouse IgG Dynal beads. Naive recipient rats (DA) were in a set of experiments in which they were given i.p. injections of 1.0 mg of the anti-rat mAbs (anti-IFN- (clone DB-1), anti-RT1B (clone OX-6), anti-RT1D (clone OX-17), and anti-TNP (clone Hy2.15)) in 1.0 ml of PBS-D 3 h before transfer or given s.c. injections of 0.2 mg of Etanercept (Enbrel; Wyeth Ayerst Pharmaceuticals) in 1.0 ml of PBS-D. Additionally, 20 x 106 pristane-primed spleen cells from DA rats were transferred to irradiated (600 rad) DA rats congenic for the MHC region (i.e., syngenic a/a, semiallogenic a/u, and allogenic u/u MHC alleles).
Statistics
Comparisons of incidences were evaluated by Fisher’s exact test, and the arthritis score and cytokine measurements were evaluated with the Mann-Whitney U test or Kruskal Wallis test. In all experiments, p < 0.05 was considered significant.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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T cells in PIA synovia
After an intradermal injection of pristane in DA rats, both CD4+, CD8+ T cells and B cells significantly expand in draining lymph nodes until day 6 after injection, whereafter the CD4+ T cells continue to expand until day 12 after injection, as shown in Table I. In the spleen, however, the expansion of B cells ceases at day 6 after injection, followed by a significant reduction in both T and B cells at day 12 after injection (Table I). To identify the cells infiltrating the arthritic joints, we collected arthritic synovia and peripheral blood from rats 14 days after pristane injection. Phenotypic analysis of synovial infiltrating cells and peripheral blood were performed by flow cytometry, with gates set to include total leukocytes (LCA+ cells), TCR+ cells, and CD4+ TCR+ cells. As shown in Fig. 1A, the arthritic snyovia is characterized by infiltrating lymphocytes, most of which are mainly highly activated CD4+ T cells, expressing ICAM-1, IL-2R (CD 25), RT-1D, and RT-1B (Fig. 1, A and B). This does not reflect the overall cell distribution after pristane injection, in which CD4+ T cells are only slightly increased in peripheral blood. After demonstrating the selective accumulation of highly activated CD4+ T cells in arthritic joints, we next investigated whether T cells from pristane-treated rats could transfer arthritis.
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T cellsThe arthritogenic capacity of cells from peripheral lymphoid tissue was investigated by adoptive transfer of spleen and inguinal lymph node cells collected from pristane-injected donor rats. Initially, an optimal transfer protocol was developed in which cell number, donor organ, and transfer day were taken into account (Table II). Both cells from the spleen and inguinal lymph nodes transferred PIA after reactivation by Con A in vitro. At day 7 after pristane injection, both spleen and lymph node cells could transfer a mild form of arthritis. Fourteen days after pristane injection, however, and despite the fewer lymphocytes in the spleen, these cells were more arthritogenic than cells from the inguinal lymph nodes. Moreover, recipient rats receiving 20 x 106 pristane-primed spleen cells developed more severe arthritis than rats receiving 5 x 106 spleen cells (Table II).
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3–4 wk, depending on the number of cells injected.
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TCR mAb (Table III). Spleen cells that were only pristane primed in vivo or only stimulated with Con A in vitro could not transfer arthritis, whereas cells that were both primed with pristane in vivo and reactivated in vitro, either by Con A or with plate-bound mitogenic anti-TCR mAb, could transfer arthritis. Arthritogenic cells also needed to be viable to transfer arthritis, because irradiated (1500 rad) pristane-primed and Con A-activated spleen cells could not transfer the disease. To identify the cell subset responsible for arthritis development, we proceeded by sorting subfractions of pristane-primed spleen cells before adoptive transfer.
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T cells alone transfer PIA
Cell-dose titration showed that only 5 x 106 total spleen cells, delivered i.v., were enough to provoke arthritis with 100% incidence. Spleen cells collected 14 days after injection were further subdivided into CD4+ and CD8+ T cells or CD4–, CD8–, CD25–, and CD134– fractions before Con A stimulation using magnetic selection. It was shown that CD4+, but not CD8+, T cells as well as CD8–, but not CD4–, spleen cells could transfer arthritis with an incidence of 100%. Furthermore, it was observed that both CD25– and CD134– fractions could become arthritogenic after Con A stimulation (Table IV). In conclusion, only CD4+T cells transfer PIA, and these cells do not have to be CD25+ or CD134+ before reactivation in vitro to transfer disease. We then proceeded by investigating the clonality of the arthritogenic cells and whether they were restricted to MHC class II.
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T cells that transfer PIA are not monoclonal
Pristane-primed spleen cells were reactivated in vitro with plate-bound mitogenic anti-rat TCR mAbs with specificities for various TCR alleles (i.e., anti-TCRV4, anti-TCRV8.5, anti-TCRV10, and anti-TCRV16), using Hy2.15 (anti-TNP) as an IgG1 isotype control. As shown in Table V, anti-TCRV4, anti-TCRV10, and anti-TCRV16 mAbs, but not anti-TCRV8.5 mAb, were able to activate cells to become arthritogenic. These data show that several, but not all, clonal specificities of T cells are arthritogenic.
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The role of MHC class II was examined in adoptively transferred arthritis by blocking MHC class II DQ (formerly denoted RT1B in the rat) and DR (formerly denoted RT1D in the rat) molecules with anti-RT1B and anti-RT1D mAbs, respectively (Fig. 3, A and B). Anti-RT1B (OX-6, 1.0 mg) and anti-RT1D (OX-17, 1.0 mg) mAbs were given i.p. 3 h before adoptive transfer of 20 x 106 Con A-stimulated spleen cells, using Hy2.15 (anti-TNP) as an IgG1 isotype control. Both anti-RT1B (Fig. 3A) and anti-RT1D (Fig. 3B) significantly reduced arthritis severity, whereas anti-RT1D treatment significantly reduced both arthritis incidence and severity. To exclude the possibility that the anti-RT1B and anti-RT1D mAbs could result in depletion of adoptively transferred class II-expressing T cells, we pretreated pristane-primed and Con A-stimulated spleen cells before transfer with either anti-RT1B or anti-RT1D, using anti-RT1A-stained cells as a positive control. No differences were observed between groups given arthritogenic cells pretreated with anti-RT1B (OX-6), anti-RT1D (OX-17), or anti-RT1A (C3) mAbs (Fig. 3C).
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T cells. Equal volumes of collected blood from each individual were stained with anti-TCR mAb and aquired for 60 s at constant pressure by flow cytometry. As shown in Fig. 4B, allogenic cell transfer did not affect the number of viable T cells indicating no host-vs-graft activity. Together, these data show that the arthritogenic T cells are MHC class II restricted and that both DQ (RT1B) and DR (RT1D) molecules are involved.
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To investigate the fate of the CD4+ TCR+ cells in adoptively transferred arthritis, we used (DAxDA.1I)F1 as donor rats, expressing both a and i alleles of the MHC class I RT1A and the MHC class II RT1B and RT1D molecules, and irradiated DA.1I as recipient rats, only expressing the i allele of these molecules, to follow cells in vivo. By doing so, we obtained a cell-target system in which we avoid both graft-vs-host as well as host-vs-graft rejection, and at the same time, we could use a mAb against the a haplotype of the RT1A molecule to detect donor cells. As depicted in Fig. 5A, arthritic DA.1I recipient rats show a severe pannus formation in ankle joints at day 12 after transfer, in which RT1A a-haplotype donor cells are located among RT1A i-haplotype nucleated recipient cells. Synovial and lymphoid organ cells from the recipient rats were isolated for flow cytometry, the gate was set to include RT1A a-haplotype donor cells (Fig. 5B), and the tissues were analyzed for CD4 expression (Fig. 5C). Fig. 5D shows that the donor cells are located more frequently in the recipient synovia than in blood, inguinal lymph nodes, and spleen. In all tissues examined, >96% of the donor cells were T cells. However, in the synovia, 99% of these T cells were CD4+, in contrast to lymphoid tissues that had a significantly lower percentage of CD4+ cells. We conclude that the transferred CD4+ T cells reached the joints and directly caused arthritis. The next question we asked was how these cells mediated arthritis.
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and TNF- production has a pivotal role for arthritis development
To investigate the functional properties of the arthritogenic T cells, we first determined their cytokine profile. Spleen cells primed in vivo with pristane and restimulated in vitro with or without Con A were examined for the production of IFN-, TNF-, and IL-4 (Table VI). Splenocytes from both normal and pristane-primed rats produced high amounts of IFN- and TNF-, but not IL-4, when restimulated in vitro with Con A. No cytokine production was detected in medium from non-restimulated cells.
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and TNF- in adoptively transferred arthritis, we pretreated naive recipient DA rats with blocking amounts of anti-IFN- (DB-1) or with Etanercept (recombinant TNF- receptor) before cell transfer. Recipients were either given i.p. injections of 1.0 mg of anti-IFN-, using 1.0 mg of Hy2.15 (anti-TNP) as an IgG1 isotype control, or s.c. injections of 0.2 mg of Etanercept (in 1.0 ml of PBS-D), using 1.0 ml of PBS-D as a control, 3 h before transfer. As shown in Table VII, both the anti-IFN- and the Etanercept treatments significantly ameliorated arthritis development both regarding incidence and arthritis severity compared with control rats.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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and TNF-. This makes the PIA model useful for investigating mechanisms of the T cell-mediated arthritis. It is well known that T cells are able to transfer various adjuvant-induced arthritides. It was, in fact, first shown using the mycobacteria adjuvant arthritis model with thoracic duct lymphocytes (27), and it has later been reproduced in models such as avridine-induced arthritis (10) and also in PIA (28). Interestingly, as in RA, all of these models also show a genetic association with the MHC region (14, 29, 30), implicating a role for MHC-restricted T cells. Additional studies to reveal the MHC specificity/restriction, T cell specificity, and effector function have been hampered with a multitude of difficulties. Nevertheless, it has been reported that T cells specific for heat shock protein can transfer arthritis; the mechanism was believed to be due to cross-reactivity of the transferred T cells with various joint Ags (31). However, these studies have not been reproduced, and histological evidence showed no or possibly only very mild synovitis (32).
In arthritis models induced by an immunization with an Ag, such as CIA, the development of arthritis is dependent on a T cell response to the immunogen, often involving only one MHC class II molecule and one or a few Ag-derived peptides (33). In PIA, there is, however, no reason to assume that only one Ag or one immunodominant peptide causes the disease, and it is, in fact, not known whether MHC class II is involved at all. Therefore, the present observation that the arthritogenic T cells in PIA are MHC class II restricted makes the model more useful for studies of a MHC class II-associated but complex disease as RA. It shows that classical MHC class II-restricted T cells are of pathogenic importance and that their specificity is related to the disease process in PIA. This indicates a role for MHC class II in a model that is not induced with an exogenous immunogen. The peptides bound by the MHC class II molecule are presumably derived from endogenous Ags, although we have no clue of the nature of this Ag(s). The arthritogenic T cells were initially activated in vivo through pristane, which does not bind MHC class II but could possibly bind other cell-surface receptors (e.g., CD1) (34). However, because the arthritogenicity was dependent on MHC class II, a direct pathogenic role of CD1-restricted T cells is unlikely. The finding that arthritogenic T cells use several different TCR V-genes and, in addition, both interact with both of the major MHC class II isotypes argues that it is likely that there is not a single self-Ag or antigenic peptide recognized. Still, at some stage, the inflammatory response, and possibly also immune recognition, is likely to be joint specific because only cartilagenous peripheral joints, with a similar distribution as in RA, are attacked. A joint-specific inflammatory attack may not, however, necessarily need a joint-specific T cell recognition. This has been shown from the observations that glucose-6-isomerase, a systemically occurring protein, is involved in arthritis in mice (35). In fact, also in CIA, the arthritis distribution is more limited than the distribution of the target self-Ag, type II collagen (36). However, in both the G6PI-induced arthritis and in the CIA model, immune recognition has been shown to occur in the lymph nodes draining the joints (37, 38), suggesting that the arthritogenic immune recognition is related to joints. Thus, for the pristane-injected donor rat, it is possible that the initial Ag/MHC recognition occurs in draining lymph nodes shortly after injection where we can observe an expansion of lymphocytes. However, the situation might be more complicated because T cells derived from the spleens are more arthritogenic than those from the lymph nodes. The reason for this remains to be investigated, but it is possible that the first pristane-mediated activation of the T cells is not MHC class II restricted/Ag dependent, implying a second step of Ag activation. An alternative explanation could be that both aggressive and regulatory T cells are activated and that regulatory T cells are more dominating in lymph nodes draining the joints.
The arthritis in both PIA and the arthritis transferred by T cells from pristine-injected rats occurs only in peripheral cartilagenous joints; in fact, they are more specific than CIA (39). This is not related to where pristane or the T cells are injected but could be related to the source of the endogenous peptides recognized. We know from studies in CIA that the nature of such endogenous proteins could be quite complex because the T cell recognition in this case is dependent on various forms of glycosylation of type II collagen (40). In addition, this glycosylation is dependent on the status of chondrocytes in the joints that could vary depending on inflammatory conditions (41). In the present experiments, we could show that the T cell-induced inflammatory cascade in the joints is dependent on cytokines such as IFN- and TNF-, which are believed to be secreted by T cells and macrophages. Infiltration of mononuclear cells into the joint synovium characterizes the chronic inflammatory process in RA and results from the migration of lymphocytes and monocytes through the endothelium of postcapillary venules (42). Both IFN- and TNF- are able to activate endothelial cells allowing traffic of arthritogenic cells into the joint synovia. Blocking of TNF- in RA patients is clearly therapeutic (43), whereas blocking of IFN- gave some initial effects but had a less promising long-term outcome (44). There are certainly many different mechanisms of the arthritogenic actions of IFN- and TNF- in both RA and PIA that need to be investigated. Therefore, the present T cell-mediated arthritis model could be a useful tool to study in detail downstream effector mechanisms of TNF- and other cytokines in a T cell/self-peptide/MHC class II-mediated disease.
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Acknowledgments |
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Disclosures |
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Footnotes |
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1 This work was supported by the Crafoord, Lundberg, Kock, and Österlund Foundations, the Swedish Association against Rheumatism, the Swedish Medical Research Council, the Swedish Foundation for Strategic Research, Arexis AB, and the European Union framework 5 program (C5RD-CT-200-00267) Van ae B.T. och aer det nodvaendigt. 
2 Current address: Pharmexa A/S, Kogle Alle 6, DK-2970 Hørsholm, Denmark.
3 Current address: Arexis AB, Arvid Wallgrensbacke 20, SE-413 46 Göteborg, Sweden.
4 Address correspondence and reprint requests to Dr. Rikard Holmdahl, Section for Medical Inflammation Research, Sölvegatan 19, I11 Biomedical Center, Lund University, S-221 84 Lund, Sweden. E-mail address: rikard.holmdahl{at}inflam.lu.se
5 Abbreviations used in this paper: RA, rheumatoid arthritis; CIA, collagen-induced arthritis; PIA, pristane-induced arthritis.
Received for publication October 13, 2004. Accepted for publication October 24, 2005.
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-chain heterogeneity among interleukin 2 receptor-positive synovial T cells suggests a role for superantigen in rheumatoid arthritis. Proc. Natl. Acad. Sci. USA 88: 10921-10925. : expression in recombinant Pichia pastoris, purification, characterization and development of a novel ELISA. Eur. Cytokine Netw. 10: 383-392. [Medline]
hrlund-Richter, S. Pettersson, R. Mattsson, R. Holmdahl. 1994. Expression of a transgenic class II Ab gene confers susceptibility to collagen-induced arthritis. Eur. J. Immunol. 24: 1698-1702. [Medline] in rheumatoid arthritis. J. Rheumatol. 20: 1867-1873. [Medline]This article has been cited by other articles:
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  A. Sattler, U. Wagner, M. Rossol, J. Sieper, P. Wu, A. Krause, W. A. Schmidt, S. Radmer, S. Kohler, C. Romagnani, et al. Cytokine-induced human IFN-{gamma}-secreting effector-memory Th cells in chronic autoimmune inflammation Blood, February 26, 2009; 113(9): 1948 - 1956. [Abstract] [Full Text] [PDF]
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 C. Rintisch, J. Ameri, P. Olofsson, H. Luthman, and R. Holmdahl Positional cloning of the Igl genes controlling rheumatoid factor production and allergic bronchitis in rats PNAS, September 16, 2008; 105(37): 14005 - 14010. [Abstract] [Full Text] [PDF]
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 M. H. Hoffmann, J. Tuncel, K. Skriner, M. Tohidast-Akrad, B. Turk, S. Pinol-Roma, G. Serre, G. Schett, J. S. Smolen, R. Holmdahl, et al. The Rheumatoid Arthritis-Associated Autoantigen hnRNP-A2 (RA33) Is a Major Stimulator of Autoimmunity in Rats with Pristane-Induced Arthritis J. Immunol., December 1, 2007; 179(11): 7568 - 7576. [Abstract] [Full Text] [PDF]
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C. Beeton, H. Wulff, N. E. Standifer, P. Azam, K. M. Mullen, M. W. Pennington, A. Kolski-Andreaco, E. Wei, A. Grino, D. R. Counts, et al. Kv1.3 channels are a therapeutic target for T cell-mediated autoimmune diseases PNAS, November 14, 2006; 103(46): 17414 - 17419. [Abstract] [Full Text] [PDF]
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K. A. Gelderman, M. Hultqvist, J. Holmberg, P. Olofsson, and R. Holmdahl T cell surface redox levels determine T cell reactivity and arthritis susceptibility PNAS, August 22, 2006; 103(34): 12831 - 12836. [Abstract] [Full Text] [PDF]
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), and PIA transfer was induced in naive DA rats (n = 6) by i.v. injection of 20 x 106 pristane-sensitized and Con A reactivated spleen cells (
).
) mAbs. B, DA recipients (n = 8) were given injections of either 1.0 mg of anti-RT1D (black and white circles) or isotype control (
), anti-RT1D (
, p < 0.05 (Fisher’s exact test).
), or MHC-allogenic (n = 8; 
) recipients. B, Total number of 