Data in Table 1 are organized according to the sequence of transfers, as shown in Fig. Open in new tab To confirm the pathophysiological role of anti-PG autoantibodies, IgG fraction was isolated from the pooled sera of naive or arthritic mice using protein G-Sepharose CL-4B chromatography.
The serum IgG fraction containing 2. In reciprocal experiments, purified WT B cells naive or arthritic or serum IgG naive or arthritic were injected first and then 5 × 10 6 purified T cells were transferred with μg PG antigen. Clinical assessment of arthritis Recipient SCID mice were examined three times a week for clinical symptoms of arthritis.
Internal Medicine: Clinical Immunology - Rheumatology
The time of onset and incidence of arthritis were recorded, and the severity was scored based upon swelling and the redness of each paw, with scores for each paw ranging from 0 to 4 yielding a maximum severity score of 16 922 For histopathological examination, the limbs of arthritic and non-arthritic mice were dissected, fixed, decalcified and embedded in paraffin.
Tissue sections were stained with hematoxylin and eosin. Appropriate isotype controls were used to determine the background staining.
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Measurement of antigen PG -specific antibodies and T cell responses Sera and spleen cells were collected from SCID mice prior to cell transfer, 3 and 7 days after the second transfer and at the end of experiments.
Histology and immunohistochemistry Femoral heads were used to show the entire articular cartilage surface rheumatoid arthritis polyarthritis fixation and decalcification. Fluorescence was examined and images were analyzed using a Nikon confocal microscope and MetaMorph image processing program Meta Imaging Series, version The contralateral hip joints were fixed and embedded for conventional histology. To detect the loss of cartilage PG, serial sections of cartilage from the femoral heads were stained with safranin-O and kulcscsont artrózisa green.
After repeated washing in serum-free DMEM, 1 × 10 6 chondrocytes were treated with undiluted sera or serial dilutions, and of naive or pooled sera from arthritic mice together with a dilution of Low-Tox R -M rabbit complement Cedarlane Inc. Kenőcs kézízületekre mouse sera were complement-inactivated for 30 min at 56°C.
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Additional negative control sera from methylated BSA-immunized mice were used. This cell tracker requires active cell metabolism thus only live cells are stained. The same cell suspensions were smeared on microscope slides and were mounted with 1.
This CellTracker labeling allowed us to visualize and quantify dead and viable cells simultaneously. Statistical analysis Descriptive statistics were used to determine group means and standard errors of the fájdalom a térdben mean ± SEM unless otherwise stated. T cells transferred from naive WT mice failed to induce arthritis followed by any combination of second transfer Fig.
These observations suggested that activated T cells from arthritic TCR-Tg animals could generate an optimal milieu for PGIA development, but T cells from arthritic WT or naive TCR-Tg mice required the additional help of B cells from arthritic mice, which might be replaced with high-dose autoantibodies to achieve a successful induction of arthritis.
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The mode of second transfers same T cells, B cells or Igs; isolated from naive or arthritic mice is indicated by symbols at the left corner of Panel A. Data represent the mean values and error bars are not shown for clarity. This observation indicated the crucial role of PG-specific B cells or antibodies in the mechanisms of arthritis development.
Four weeks after the second transfer, T cell homeostasis was fully reconstituted in all SCID recipients These data suggested that a high dose of exogenous anti-PG autoantibodies or B cells from PG-immunized mice could supplement and support the relative antigen-specific B cell deficiency in these SCID mice, which would cause a milder form of transferred arthritis Fig. However, these high-dose anti-PG autoantibodies or B cells from PG-immunized mice were not sufficient to induce minimal signs of synovitis when recipient mice did not receive antigen-specific T cells Fig.
Based on these findings, we concluded that antigen specificity and activation status rather than the number of recovered T cells could orchestrate an optimal milieu for the recovery of arthritogenic B cells in recipient SCID mice. Rheumatoid arthritis polyarthritis in antigen-specific T and B cell responses controlled the success of adoptive transfer To investigate the pathogenic effect of these colonizing and proliferating B cells, we measured anti-PG antibody levels in the sera of mice after adoptive transfer.
B IL-2 secretions from the supernatants of PG-activated spleen cells were measured and are shown as stimulation indices that are normalized to medium control. Rheumatoid arthritis polyarthritis transfer of B cells followed by T cells To further investigate the sequential events in the development of adoptive transfer-induced PGIA, we performed reciprocal transfer experiments in which B cells from naive or arthritic donors were injected first into SCID mice, followed by the transfer of purified T cells together with PG antigen.
In these reciprocal transfer systems, arthritis developed only in mice that received T cells in the rheumatoid arthritis polyarthritis transfer from rheumatoid arthritis polyarthritis TCR-Tg donors Fig. When B cells were injected before T cell transfer, their number, activation status or antigen specificity did not seem to be important because only T cells from arthritic TCR-Tg donors, but not from arthritic WT donors, could induce arthritis effectively Fig.
SCID mice that had received naive B cells and naive T cells from WT mice did not develop arthritis and were negative for all measured parameters data not shown. Significant IL-2 Fig.
These reciprocal transfer experiments further emphasize the role of activated and antigen-specific T cells in the pathology of autoimmune processes. C T cell responses were measured by IL-2 production and are shown as stimulation indices normalized to medium controls. Anti-PG autoantibodies accelerated disease after adoptive transfer with arthritic T cells In contrast to B cells, pre-treatment of SCID mice with PG-specific rheumatoid arthritis polyarthritis accelerated the onset and increased the incidence of arthritis when followed by the adoptive transfer of T cells from arthritic donors Fig.
Anti-PG antibodies were measured after the second transfer and were at especially high levels when arthritic T cells from WT animals were used for the second transfer Fig. Note that the use of T cells from arthritic TCR-Tg mice was rheumatoid arthritis polyarthritis in this experimental setup because a single transfer of these T cells could transfer arthritis without the additional transfer of B cells or anti-PG antibodies see Fig.
Therefore, the serum antibodies in each of the four groups of mice Fig. C T cell responses were measured by IL-2 production and are shown as stimulation indices that are normalized to medium controls.
Rheumatoid arthritis – az izületek gyulladásos elváltozásairól
Antibodies and complement C3 bound to the surface of articular cartilage and chondrocytes in inflamed joints To test the potential involvement of these PG-specific antibodies in situ in cartilage damage, femoral head cartilages from SCID mice with or without adoptively transferred PGIA were removed rheumatoid arthritis polyarthritis cryosectioned.
Frozen serial sections were stained with safranin-O a marker of negatively charged PG molecules or immunostained for mouse IgG and complement C3. Although safranin-O staining showed only a minimal loss of surface PG from femoral head cartilage Fig. Whenever arthritis was more pronounced Fig. IgGs typically localized to large areas including the extracellular matrix in the rheumatoid arthritis polyarthritis layer of articular cartilage Fig. The co-localization of cartilage surface-bound IgG and chondrocyte-bound C3 suggested that the antibodies produced by activated antigen-specific B cells can actively contribute to irreversible cartilage destruction by the complement system in PGIA.
Sections shown in A panels show early 3—4 days old disease onset. Cartilage from femoral heads were embedded in OCT and cryosectioned.
Panel A3 is a merged image of A1 and A2, and panel B3 is a merged image of B1 and B2, which show the co-localization of cartilage surface-bound IgG and chondrocyte-bound complement C3. Antibodies in the sera of arthritic animals were cytotoxic toward syngeneic chondrocytes Because we detected a sufficient amount of cartilage-bound IgG and complement in the superficial layer of articular cartilage, we were interested in whether these antibodies are cytotoxic toward mouse chondrocytes.
Indeed, in a complement-mediated cytotoxic akut arthrosis kezelése using fluorescent CellTracker, we measured massive cytotoxic effect of sera from arthritic mice using an in vitro fluorescent viability test. Isolated chondrocytes were exposed to sera from normal or arthritic mice at various concentrations in the presence of a dilution of rabbit complement, and cell death was monitored by flow cytometry and fluorescence microscopy Fig.
These observations, together with the immunohistochemistry results Fig. Open in new tab Download slide Chondrocyte cytotoxicity using normal or arthritic mouse serum in the presence of low-toxicity rabbit complement.
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Panels underneath flow histograms show the corresponding light scatters and fluorescence images by immunocytochemistry. The chondrocytes were counterstained with DAPI, where blue cells represent dead cells. The induced inflammatory events synovitis disappear within a few days or a couple of weeks without irreversible joint damage. However, a common denominator in all these chronic and transient arthritis models is an antibody-induced synovitis, which disappears in the absence, yet becomes a chronic and progressive form in the presence, of auto antigen-specific T and B cells.
Reumatoid arthritis tünetei és kezelése
Among the above listed animal models of RA, PGIA is unique in terms of its female prevalence 31particularly in aging females This disease can be induced by transferring antigen PG -activated spleen cells into syngeneic mice, but neither B cells nor anti-PG antibodies alone are sufficient for arthritis induction in the absence of PG-specific T cells. Serum anti-PG antibody titers are the best biomarkers for arthritis development and disease progression.
However, we surprisingly found incomplete B cell recovery along with PG-specific antibodies in the rheumatoid arthritis polyarthritis of recipient SCID mice with arthritis. The increasing amounts of serum anti-PG auto antibodies could not only predict arthritis onset but also correlated with the severity of arthritis symptoms.
Although homeostatic T cell proliferation has been studied extensively 202140—42far less information is available for homeostatic B cell proliferation 12134344particularly in a lymphopenic environment. The pathogenic role of autoantibodies and B cell homeostasis in autoimmunity is of critical importance 5612—14 and these transfer experiments explored the importance of an activated antigen-specific T cell-dependent and highly specific B cell recovery in autoimmune arthritis.
The second transfer, which included T cells, B cells or anti-PG antibodies from arthritic mice, was an additional challenge for provoking arthritis.
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Similar results were found in reciprocal experiments in which the injection of anti-PG antibody was followed by a transfer of T cells from arthritic animals.
These observations suggest that neither activated T cells from arthritic WT mice nor the antigen specificity of T cells naive TCR-Tg alone were sufficient to support the clonal expansion of antigen-specific B cells and their subsequent antibody production, which could then lead to arthritis induction.
Thus, a threshold level of antigen-specific T cell number and activation is required to trigger antigen-specific B cell proliferation and subsequent antibody production.
The injection of anti-PG antibodies or B cells from arthritic mice had a similar effect on this lábízületi betegségek kezelése transfer system, suggesting that the important role for B cells at least in this case was limited to anti-PG antibody production; other B cell functions such as antigen presentation and cytokine production might be critical but not exclusively required. Anti-PG antibodies enhanced the efficacy of PGIA development following transfer, even when administered before the T cells, indicating that the presence of antigen-specific IgG was necessary for disease induction.
REUMATOID ARTHRITIS (polyarthritis): Miért deformálódnak az ujjak?
When anti-PG antibodies were injected before T cell transfer Fig. To translate these observations into the natural situation that occurs in RA, antibodies produced rheumatoid arthritis polyarthritis B cells that are receiving assistance from activated antigen-specific T cells within the proper milieu can cause synovitis, which becomes a chronic disease and leads to irreversible joint deformities in patients. Rheumatoid arthritis polyarthritis antibodies bind to the surface of cartilage, primarily recognizing the GAG side chains of PG molecules in the extracellular matrix and on the chondrocyte surface Fig.
The intra-articular activation of the complement cascade might result in complement-dependent leukocyte recruitment 46 and chondrocytolysis 47as rheumatoid arthritis polyarthritis in Fig.
The results from A nyaki osteochondrosis 2. fokú kezelése experimental transfer system have demonstrated that neither T cells nor antibodies or B cells alone are sufficient to induce destructive joint inflammation. Rather, a well-controlled and sequential activated antigen-specific T cell-driven B cell selection and activation as well as subsequent autoantibody production can lead to the cartilage destruction executed by the complement system.
Although a uniform TCR repertoire as present in TCR-Tg mice does not exist in humans, an oligoclonal cumulative effect over many years may lead to the development of a similar condition.
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The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. Competing interest The authors declare no competing financial interests. Oswald, who assisted in studies involving SCID mice. Berlo and Dr Chris P.