Immunisation of certain rodent strains, of particular MHC types with heterologous type I1 collagen (CII) results in the development of an acute arthritic disease called collagen-induced arthritis (CIA) [l]. The disease is thought to result from anticollagen antibody immune complex-mediated development of synovitis and macrophage-mediated erosive cartilage destruction. The disease has also been shown to be T cell dependent since T cell depleted animals are not susceptible to arthritis [2] and the disease can be successfully treated with anti-CD4 [3] and anti-TCR monoclonal antibodies [4]. Significantly, T cell proliferative responses have been shown to immunodominant epitopes of CII, located particularly in the cyanogen bromide (CB) fragment CB 1 1 [5,6]. Intradennal immunisation of Lewis rats (MHC RT1-1) with bovine CII (KII) leads to an acute inflammatory response from day 14 post immunisation. In order to determine the phenotype and functional properties of T cells reactive with epitopes of CII implicated in the pathology and regulation of CIA, the generation of CII specific T cell lines and clones in Lewis rats was embarked upon.Draining lymph node cells (LNC) from bCII immunised rats were harvested at day 14 and set up in bulk culture with bCII (2Opglml) in RPMI supplemented with 2% normal rat serum. At day 4 massive proliferative responses (CPM=90,358) of whole LNCs in the presence of bCII were observed (SI=26). Characterisation of these responding cells by FACscan analysis showed an expanding population of CD4+. MHC II+, IL-2 receptor+ and CD45RC-T cells. In order to determine suitable antigen presenting cells ( A X ) for presentation of bCII at restimulation a proliferation assay was also set up with separated T cells and different sources of Apc. T cell proliferation and SI was greatest with a ratio of 1 T cell : 20 thymocytes, normal LNC gave a poor SI and splenocytes were found to be inhibitory. At day 12 of bulk culture, when the cells were rested, viable cells were harvested and restimulated with thymocytes pulsed overnight with or without bCII. Following restimulation a large proliferative response to the addition of thymocytes alone was observed within 24 hours, peaking at 48 hours (Fig. 1). . . Fig. 1 u s AFC is Responder cells were harvested from bCII stimulated cultures at day 12 and restimulated with bCII-pulsed or non-pulsed, mitomycin-C treated thymocytes. Responder cells were used at 5x104/well and were plated out in triplicate. Tritiated thymidine was added for the final 6 hours of culture and the assay was harvested at 48 hours.Abbreviations used CIA-collagen-induced arthritis, CII-type I1 collagen, LCII-bovine type I1 collagen, LNC-lymph node cell, APC-antigen presenting cell, OVA-ovalbumin.This autoreactive response increased with increasing numbers of thymocytes added and was also seen with splenocytes and normal LNCs as AFC. Numerous changes to the protocol e.g. the omission of IL-2 to reduce bystander expansion in order to try and generate CII specificity were unsuccessful; autoreactiv...
The structure of alpha beta T cell receptors (TCR) is restricted in a number of rodent and human antigen responses. In several rodent EAE models of multiple sclerosis a limited range of T cell receptors are expressed by T cells which respond to the inciting antigen and are capable of transferring the disease to naive animals. These observations have raised the question of whether in rheumatoid arthritis (RA), a particular T cell receptor structural signature can be identified among T cells derived from the synovium compared to autologous peripheral blood. The parameters which are usually measured are TCR variable region usage, oligoclonality and/or limited junctional region usage. A large number of studies have been carried out and results are variable with some authors claiming evidence for the effect of uncharacterised superantigens expanding or deleting T cells with particular V beta regions while others have suggested that observations of restricted V region usage and limited junctional regions imply that clones of cells have been expanded by antigen. So far none of these studies have led to the identification of an antigen or superantigen which plays a role in RA pathogenesis.
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