The clinical and immunogenetic features of patients with autoantibodies to the nucleolar antigen PM-Scl.

The clinical and laboratory features of 32 patients with anti-PM-Scl were studied. Patients with this rare autoantibody suffered from a homogenous overlap connective tissue disease defined by Raynaud phenomenon (32/32), features of scleroderma (31/32), arthritis (31/32, erosive in 9/32), myositis (28/32), lung restriction (25/32), calcinosis (15/32), and sicca (11/32). Significant renal and neurologic involvement was uncommon. All patients examined (22/22) had HLA-DR3, and 50% of these patients were homozygous. Our patients responded favorably to moderate immunosuppression and, with therapy, the disease generally has a good prognosis; over 50% of our series (17/32) remained well on minimal or no immunosuppression after a median follow-up of 8 years.

Co-expression of human T cell receptor chains with mouse CD3 on the cell surface of a mouse T cell hybridoma.

In this study we demonstrate that human T cell receptor (TcR) chains can be co-expressed with murine CD3 on the cell surface of a murine T cell hybridoma. Human TcR alpha and beta genes from the Jurkat leukaemic cell line were transfected into a TcR-negative mouse T cell hybridoma, TG40. The Jurkat TcR was successfully co-expressed at the cell surface with mouse CD3 components. Brightly staining transfectants were selected by fluorescence-activated cell sorting, and levels of expression comparable to a normal T cell line were achieved suggesting that the human TcR dimer assembled efficiently with the mouse CD3 complex. Northern blot analysis demonstrated similar levels of TcR messenger RNA to those found in the parental Jurkat line. Although we have not formally demonstrated surface expression of the Jurkat TcR alpha chain, the residual alpha gene transcript which is present in murine TG40 line is non-expressible. In order to test the signalling capacity of this human/mouse complex, the transfectants were stimulated with an anti-V beta 8 monoclonal antibody. This stimulus led to interleukin-2 production by the transfectants, demonstrating the delivery of a transmembrane signal. In addition, B10.A mice were immunised with the transfectants, and the antisera from these mice stained the transfectant and the Jurkat cell line, but not the parental T cell hybridoma. This interspecies transfection approach should now permit us to explore the requirements for T cell activation, the constraints on TcR alpha beta chain pairing, and creates ideal reagents for inducing a mouse anti-human TcR-specific response with a view to producing panels of anti-human TcR monoclonal antibodies.

Anti-PL 4 in patients with systemic lupus erythematosus with severe renal and haematological disease.

Anti-PL 4 is an autoantibody which binds to a 150 kDa polypeptide and is found in approximately 1% of SLE sera. The clinical and laboratory features of 16 patients who have had anti-PL 4 detected in their serum are presented. Anti-PL 4 is highly specific for SLE (100%) and identifies a population of patients who typically develop severe renal (75%) and haematological disease (100%).

[Occupational eosinophilic lung in a grape grower: role of sulfites]. / Poumon éosinophile d'origine professionnelle chez un viticulteur: rôle des sulfites.

Observation of a patient with a history of asthma who presented a lung pathology in a context linked with the general condition, radiology of a biological inflammatory syndrome and hypereosinophilia, suggested a diagnosis of Chronic Pneumopathy of Eosinophils (PCE). The chronology of the symptomatology favours an occupational role of sulphites, to which the patient is exposed by the respiratory pathway and the reproduction of the symptoms in an identical exposure confirmed the hypothesis.

PCR-based analysis of the TCR repertoire in human autoimmune diseases.

Characterization of T cells at sites of autoimmune damage has been difficult. Now, however, polymerase chain reaction (PCR)-based methods are being used to analyse T-cell receptor (TCR) gene expression in such lesions. Here, Christopher Marguerie, Claudio Lunardi and Alex So summarize and interpret the most recent findings and describe the current understanding of TCR usage in autoimmune diseases.

Identification of novel human T-cell receptor V beta gene segments by the anchored-polymerase chain reaction.

We have studied the diversity of the expressed human T-cell receptor (TCR) beta-chain repertoire by analysis of mRNA from unstimulated peripheral blood T-cells. The anchored-polymerase chain reaction (PCR) was used to isolate TCRB transcripts. Of 20 full or near full-length functional transcripts sequenced, two were novel TCRVB gene segments. They have strong sequence similarities to the known TCRBV5, and 8 subfamilies. Southern blot analysis and sequence-specific oligonucleotide hybridization confirmed: a) that these sequences are present in genomic DNA; b) their relationship to the known TCRBV families. A TCRBV sequence similar to a recently identified novel TCRBV24 subfamily was also found. We show by southern blotting that this sequence forms a single member subfamily, and by deletion analysis of T-cell lines, we have mapped this sequence to lie between the genes which encode the TCRBV8.1 and TCRBV5.3 gene segments. The results show that the anchored PCR is a powerful tool in the analysis of the TCR repertoire, which may contain more V gene segments than previously defined.

An altered repertoire of T cell receptor V gene expression by rheumatoid synovial fluid T lymphocytes.

The pattern of T cell receptor V gene expression by lymphocytes from rheumatoid synovial fluid and paired peripheral blood samples was compared using a polymerase chain reaction (PCR)-based assay. Eight rheumatoid arthritis (RA) patients who had varying durations of disease (from 2 to 20 years) were studied. In all patients there was evidence of a different pattern of V gene expression between the two compartments. Significantly increased expression of at least one V alpha or V beta gene family by synovial fluid T cells was observed in all the patients studied. Three different V alpha (V alpha 10, 15 and 18) and three V beta (V beta 4, 5 and 13) families were commonly elevated. Sequencing of synovial V beta transcripts demonstrated that the basis of increased expression of selected V gene families in the synovial fluid was due to the presence of dominant clonotypes within those families, which constituted up to 53% of the sequences isolated from one particular synovial V gene family. There were considerable differences in the NDJ sequences found in synovial and peripheral blood T cell receptor (TCR) transcripts of the same V beta gene family. These data suggest that the TCR repertoire in the two compartments differs, and that antigen-driven expansion of particular synovial T cell populations is a component of rheumatoid synovitis, and is present in all stages of the disease.

T gamma delta cells and their subsets in blood and synovial fluid from patients with rheumatoid arthritis.

We have determined the distribution of T gamma delta cells in the peripheral blood of 44 patients with rheumatoid arthritis and in 36 healthy controls. In addition, paired blood and synovial fluid samples were obtained from seven patients with RA. The monoclonal antibodies A13, BB3 and Ti gamma A, which are specific for the V delta 1, V delta 2 and V gamma 9 gene products respectively, were used to define T gamma delta subsets. T gamma delta + cells expressed as a percentage of CD3+ lymphocytes were reduced in RA peripheral blood compared with the control group (3.9% +/- 0.5 versus 5.7% +/- 0.7; P less than 0.0001). There was a reduction in the V gamma 9/V delta 2+ subset (from 5.6% +/- 1.2 to 1.7% +/- 0.4) leading to a change in the mean ratio of V delta 2/V delta 1+ cells from 4.3 in normal subjects to 1.1 (P less than 0.002). No statistical difference was observed in T gamma delta cell numbers in synovial fluid compared with the paired blood samples (4.0% +/- 1.1 in blood and 4.4% +/- 1.4 in synovial fluid). Also the distribution of V delta 2+ and V delta 1+ cells was similar in the two compartments and a similar alteration in subset distribution was found in blood and synovial fluid. These findings do not indicate a selective accumulation of a specific T gamma delta subset in RA synovial effusions.

Reduction in T gamma delta cell numbers and alteration in subset distribution in systemic lupus erythematosus.

We have studied the distribution of T gamma delta cells in the peripheral blood of 35 patients with systemic lupus erythematosus (SLE) and 36 age-matched controls. The monoclonal antibodies A13, BB3 and Ti gamma A, which are specific for the V delta 1, V delta 2 and V delta 9 gene products respectively, were used to define T gamma delta cell subsets. A significantly lower frequency of T gamma delta cells was found in peripheral blood lymphocytes of SLE patients compared with normal subjects (3.2% versus 5.9%). There was a marked reduction in the V delta 2+ subset of T gamma delta cells, which resulted in a reversal of the ratio of V delta 2+/V delta 1+ cells from 4.34 to 0.56. No correlation was found with either clinical or laboratory measures of disease activity. These results suggest that the observed changed in T gamma delta subset distribution are related to the SLE itself, and not secondary to changes in disease activity.

Malabsorption caused by coeliac disease in patients who have scleroderma.

Coeliac disease may account for malabsorption in scleroderma patients even when test suggest bacterial overgrowth. A small bowel biopsy is essential.

The genes coding for A alpha-, B beta-, and gamma-chains of fibrinogen map to 4q2.

We used cloned cDNA probes for the A alpha-, B beta-, and gamma-chains of human fibrinogen and Southern blotting techniques to analyze DNA from a series of rodent X human somatic cell hybrids for the presence of specific fibrinogen-related sequences. Our results provide evidence for the assignment of the three genes for fibrinogen to chromosome 4. Moreover, by direct gene-dosage determination in two patients with chromosome 4 unbalanced rearrangements, we refined the regional chromosomal assignment to 4q2, thus suggesting that these three genes whose expression is coordinately regulated are closely linked.

Polymyositis, pulmonary fibrosis and autoantibodies to aminoacyl-tRNA synthetase enzymes.

The clinical and laboratory features of 29 patients who had one of three anti-aminoacyl-tRNA synthetase autoantibodies, anti-Jo1 (histidyl-tRNA synthetase), anti-PL12 (alanyl-tRNA synthetase) or anti-PL7 (threonyl-tRNA synthetase) were analysed and compared with the findings of other published reports. These autoantibodies were found to be associated with a syndrome delineated by inflammatory myositis (24 patients) and pulmonary fibrosis (23 of 29), but also including inflammatory arthritis (26/29), keratoconjunctivitis sicca (17/29), sclerodactyly (21/29), Raynaud's phenomenon (27/29), hepatitis (8/29) and subcutaneous calcinosis (7/29). The most important clinical determinant of outcome in this group of patients was the severity of the interstitial pulmonary disease. No patient fulfilled the classification criteria for systemic lupus erythematosus, although 10 had autoantibodies to extractable nuclear antigens including Ro, La, RNP, and Sm, and two patients had anti-dsDNA antibodies. Although it seems unlikely that anti-aminoacyl-tRNA synthetase antibodies are directly responsible for causing disease, they may provide an important clue to the aetiology of this unusual syndrome.