T cell activation in the characteristic synovial lesions of rheumatoid arthritis may play a major role in the pathogenesis of this autoimmune disease. Analysis of T cell clonal diversity in these sites remains equivocal. Using the PCR and subsequent single-strand conformation polymorphism analysis it is possible to assess the degree of junctional diversity in the TCR with minimal selection bias. Concentrating on the beta-chain of the TCR, a paucity of clonotypic T cell expansion is demonstrated in the peripheral blood of healthy individuals. After polyclonal stimulation in vitro (with concanavalin A or phytohemagglutinin) this pattern does not change. In contrast, some T cell clonotypes appear following in vitro stimulation with purified protein derivative. Analysis of the peripheral blood, synovial fluid, and synovial tissue of patients with rheumatoid arthritis indicated many dominant T cell clonotypes. These data argue for a clonally diverse T cell response in the affected tissues of rheumatoid arthritic subjects.

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In an attempt to identify autoantigens of synovium in rheumatoid arthritis (RA), we constructed lambda phage expression cDNA libraries from synovium and screened them by IgG purified from synovial fluids, both of which were derived from RA patients. As a result of this unique combination of the libraries and probes, we cloned follistatin-related protein (FRP) as a novel autoantigen in systemic rheumatic diseases. FRP is a secreted protein containing a similar amino acid sequence to follistatin, an inhibitor of activin. FRP was first cloned as a transforming growth factor-beta1-inducible protein (called TSC-36) from a mouse osteoblastic cell line and was suggested to have some roles in the negative regulation of cellular growth. Immunoblotting analyses detected synovial fluid and serum anti-FRP antibodies of IgG class more frequently in RA than any other systemic rheumatic diseases and controls. Synovial fluid anti-FRP antibodies appeared in 44% of RA (n = 18) and none of osteoarthritis (OA) (n = 15) patients. Serum antibodies were detected in 30% of RA (n = 67), 17% of systemic sclerosis (n = 18), 10% of systemic lupus erythematosus (n = 51) and Sjögren's syndrome (n = 10), and none of polymyositis/dermatomyositis (n = 13) patients and healthy subjects (n = 30). These antibodies recognized an EC domain, an extracellular Ca2+ binding module. In anti-FRP antibody-positive RA patients, serum C-reactive protein level and erythrocyte sedimentation rate were more elevated than negative patients (P < 0.05 and P < 0.01, respectively). FRP gene expression was higher in RA than OA synovium (P < 0.05). However, there was no difference between these groups in the amount of synovial FRP, suggesting its elevated turnover in RA. As follistatin inhibits activin, FRP might inhibit some growth factor-like molecule. Detection of anti-FRP antibodies, possibly having disease-promoting effects as the blocking antibodies, could be one of the markers for clinical evaluation of systemic rheumatic diseases.

The enhanced oncogenicity of particular human papillomavirus type 16 (HPV16) E6 variants is population-dependent, implying the involvement of additional genetic cofactors. This study was designed to investigate the association between E6 variants and human leukocyte antigen (HLA) polymorphism within a Japanese population. Fifty-seven women with HPV16-positive cervical cancer were analyzed for E6 sequence variation and its relationship to HLA class II alleles. Compared with local controls (n ‫؍‬ 138) and published controls (n ‫؍‬ 916), DRB1*1501 and DQB1*0602 frequencies were significantly increased among patients with HPV16 E6 prototype (n ‫؍‬ 11). Additionally, DRB1*1502 was positively associated with a particular E6 variant designated D25E (n ‫؍‬ 25), although we could not find a significant association between HLA class II alleles and L83V variants (n ‫؍‬ 16). Infections by oncogenic human papillomaviruses (HPVs) are established as a major risk factor for cervical cancer. 1 HPV16 is the most frequently detected papillomavirus in invasive cervical cancer worldwide. HPV16 E6 and E7 proteins are known to inactivate the function of oncosuppressor proteins p53 and pRb, respectively, and immortalize human keratinocytes in vitro. 2 Recently, certain HPV16 E6 variants having Ͻ 2% sequence variation compared with the HPV16 prototype strain have been reported to confer an increased risk for cervical cancer in a given population. [3][4][5][6] Interestingly, the oncogenicity of specific E6 variants differs between various ethnic populations. 7 The E6 L83V variant, which has valine (V) substituted for leucine (L) at residue 83, represents a higher risk for cervical cancer in a Swedish population. However, the L83V variant does not carry a significant risk in either Italian or Czech populations. Population-related oncogenicity of HPV16 carrying specific E6 variations suggests the involvement of additional genetic cofactors, for instance human leukocyte antigen (HLA) types. Additionally, there are data suggesting that HLA association with cervical cancer shows HPV type specificity. 8 However, little is known about a possible link between HPV intratypic variations and HLA types.To investigate the role of HLA polymorphism among HPV16 E6 variants in cervical carcinogenesis, we analyzed HPV16 E6 sequence variation and its relationship to HLA class II alleles in 57 Japanese women with HPV16-positive cervical cancer. Since there is little evidence suggesting a possible association between HLA class I alleles and cervical cancer, the present study focused on HLA class II alleles. Although the sample size of the present study was small, we found some significant associations between E6 variants and HLA class II alleles. MATERIAL AND METHODS Study subjectsAmong 161 women who underwent treatment for primary cervical cancer at University of Tokyo Hospital and Saitama Cancer Center between 1997 and 2001, 57 Japanese women with HPV16-positive cervical cancer [6 carcinoma in situ (CIS) and 51 invasive cervical cancer (ICC)] were tested f...