The induction of human antinuclear antibodies by D-penicillamine: activation of inducer helper T cells in the absence of irradiation sensitive suppressor T cells.

The capacity of D-Penicillamine (DP) to induce or to potentiate the production of antinuclear antibodies (ANA), detected by immunofluorescence (IF), was investigated in vitro, using peripheral blood mononuclear cells (PBMC) from patients with systemic lupus erythematosus (SLE) and normal individuals. Except in one patient with SLE, DP did not enhance ANA synthesis when using unseparated PBMC. In contrast, when B cells were cocultured with irradiated T cells or irradiated enriched T4+ subset, DP induced or potentiated the production of ANA. These results indicate that DP acts by stimulating T4+ helper cells to promote ANA synthesis in the absence of radio-sensitive suppressor T cell function contained within the T4+ population.

D-penicillamine: a modulator of anti-DNA antibodies production.

The effect of D-Penicillamine (DP) on the in vitro production of anti-DNA antibodies by peripheral blood mononuclear cells (PBMC) from patients with systemic lupus erythematosus (SLE) and from healthy individuals was studied. Anti-DNA antibodies were measured in culture supernatants using a sensitive microenzyme-linked immunoassay technique. The results of this investigation suggest that DP can act as an immunomodulator capable of potentiating or initiating anti-DNA antibodies synthesis as well as suppressing it. Although PBMC from both SLE patients and controls were responsive to this thiol compound, our results indicate that PBMC from patients with SLE were more susceptible to the enhancing effect of DP than did PBMC from controls. The cellular mechanism by which this drug can modulate anti-DNA antibodies production is discussed.

The in vitro production of anti-nuclear antibodies by human peripheral blood mononuclear cells. Demonstration of T cell requirement and soluble inducing factor(s) for anti-nuclear antibodies triggering in patients with systemic lupus erythematosus.

Peripheral blood mononuclear cells (PBMC) of 29 patients with systemic lupus erythematosus (SLE) and 14 normal individuals were investigated for the in vitro production of anti-nuclear antibodies (ANA). Twenty-eight of 29 SLE patients but only one control spontaneously produced ANA in unstimulated PBMC. Pokeweed mitogen induced ANA synthesis in six controls. No detectable ANA was observed in B cell enriched fraction except in two cases of SLE. Recombination of B + T cell enriched fractions and PBMC supernatants from SLE patients could induce B cells to synthesize ANA. These results indicate that: (1) SLE patients spontaneously produced ANA in vitro whereas controls rarely did; (2) autoreactive clones exist in normal individuals but are kept under control and (3) T cell help is required for ANA triggering.

The in vitro production and regulation of anti-double stranded DNA antibodies by peripheral blood mononuclear cells from normals and patients with systemic lupus erythematosus.

The in vitro production of anti-double stranded DNA antibodies (anti-DNA) by peripheral blood mononuclear cells (PBMC) was investigated in 19 patients with systemic lupus erythematosus (SLE) and in 12 normal individuals, using a micro solid phase enzyme immunoassay. PBMC from SLE patients spontaneously produced anti-DNA with a higher frequency (16 of 19) than did PBMC of controls (three of 12). In addition SLE patients produced predominantly IgG antibodies. PWM and DNA enhanced anti-DNA synthesis is spontaneously low and non-producers, but acted as inhibitors in spontaneously high producers. The partial removal of T cells decreased or abolished anti-DNA synthesis in four of nine SLE patients. In contrast the B cell enriched fractions of five of nine SLE and five of seven normal patients produced the same or higher anti-DNA levels than did the corresponding unseparated PBMC. These results suggest evidence for autoreactive B cells in SLE as well as in normals, and therefore the combination of these autoreactive B cells with helper and/or suppressor T cell disorders could lead to the over production of anti-DNA seen in different patients with SLE.

Manganese-containing superoxide-dismutase deficiency in polymorphonuclear leukocytes of adults with rheumatoid arthritis.

Superoxide dismutase (SOD) is known to regulate the level of superoxide radicals inside cells. The purpose of this work was to investigate the role of SOD activity in tissue damage produced by superoxide radicals. SOD was measured in polymorphonuclear cells of patients with rheumatoid arthritis and controls. The distinct SOD activities, including manganese-containing and copper-zinc-containing enzymes, were evaluated in cytoplasma and mitochondria of human granulocytes. Except for the comparison between total SOD and cytoplasmic copper-zinc SOD, no correlation was found among the different SOD levels. Moreover, a significant decrease was observed only for cytoplasmic manganese-containing enzyme in granulocytes of adults with rheumatoid arthritis. These data confirm the necessity of evaluation of various SOD classes and suggest the interest of biochemical tests in granulocytes for early diagnosis and better comprehension of tissue damage due to inflammation.

[Antinuclear antibodies: analysis of images and classification of immunofluorescence aspects on nuclei isolated from rat hepatocytes]. / Anticorps anti-noyaux: analyse d'images et classification des aspects d'immunofluorescence, sur noyaux isolés d'hépatocytes de rat.

Image analysis and Fourier transformation of the various nuclear immunofluorescence patterns observed while detecting antinuclear antibodies allow an objective and quantitative definition of the fluorescence. They also point out various IF types hidden by the main pattern, without having to dilute the test serum. They make obvious the difference between speckled and reticular patterns, and reveal the existence of intermediate states. The usual nuclear IF patterns (homogeneous, ring, nucleolar, reticulated, speckled and diffuse) may be grouped, according to their photo emission, into nuclear and subnuclear patterns. The first group includes homogeneous, annular and passive nucleolar IF. The second group is composed of speckled, reticulated, mixed, and active nucleolar IF. Alternatively, these aspects may be grouped into three types: homogeneous nuclear IF (homogeneous and ring), heterogeneous nuclear IF (speckled, reticulated and mixed) and nucleolar IF (active or passive). Diffusion can affect or not these aspects and does not apply to a special type or pattern. Image analysis and the study of the image spatial spectrum lead to automated recognition of the IF types, and later on, to the discrimination of antinuclear antibodies.

T suppressor lymphocytes regulation of adjuvant arthritis in two inbred strains of rats.

Adjuvant arthritis can be induced by a single injection of Freund's complete adjuvant (FCA) in the highly susceptible Lewis (LEW) rat strain, but not the resistant Wistar A.G. (WAG) strain. This strain-dependent susceptibility to the disease is correlated with differences in T suppressor cells regulation. In WAG rats, indeed, the in vitro response in LEW alloantigens was highly inhibited 11 days after FCA injection, while LEW rats in vitro response to WAG alloantigens was slightly increased. Furthermore, spleen cells from WAG rats given FCA 4 days before exhibited T cell-mediated active suppression of WAG in vitro response to LEW alloantigens when they were co-cultured with WAG normal spleen cells. This suppression was abolished by removal of T cells on nylon wool column. A previous irradiation of these T cells also inhibited their suppressive effect, suggesting that FCA-induced suppression might be due to soluble suppressor factor(s). On the other hand T cells from FCA treated LEW rats did not produce any modification of LEW in vitro response to WAG alloantigens. This suggests that the severe arthritis induced in LEW rats could be correlated with a defect of their suppressor cells functions, while in WAG rats FCA activated suppressor T cells could control the disease.

Comparison of effects of D. Penicillamine and dexamethasone on the articular and non articular lesions in immune connective tissue disease induced in rabbits.

The effect of D. Penicillamine (DP) at the dose of 50 mg/kg/day, on an immune induced connective tissue disease in rabbit, is compared to that of dexamethasone (Dexa) at the doses of 0.15 and 0.075 mg/kg/day. This model includes polyarthritis and lesions of connective tissue of liver, kidneys and lungs. The result of immunization is initially a non-specific macrophage infiltration and secondarily a specific lymphocyte and plasma-cell infiltration. In short treatment, high dose of Dexa inhibits the non-specific and specific responses while DP modifies only non specific response. In long treatment, Dexa at low dose and DP inhibit the two responses. Data suggest that, in vivo, macrophages is the target cell of DP.

Genetic control of antinuclear antibodies in mice inoculated with the moloney leukaemia virus.

The production of antinuclear antibodies (ANA) was studied after inoculation with Moloney leukaemia virus (M-MuLV) in different H-2 congenic strains of mice. Using a new sensitive method for ANA detection, it was demonstrated that M-MuLV-induced ANA were genetically controlled by several different factors. A high viral production was first required for ANA triggering. Among viremic animals both high and low ANA producers were observed. H-2 and non H-2-linked genes were involved in the control of M-MuLV-induced ANA; these genes were different from those involved in the control of viremia. The H2b haplotype was associated with an increased ANA response, the transmission of the responder phenotype being intermediate. Non-H-2-linked genes must also control M-MuLV-induced ANA, as demonstrated in mice having the same H-2 haplotypes, since with equivalent viremias they produced different amounts of ANA. No linkage with X chromosome was found.

Hidden anti-nuclear antibodies in rheumatic diseases.

Hidden anti-nuclear antibodies are demonstrated by immunofluorescence using smears of rat nuclei as substrate and rat liver section technique when sera are incubated with penicillamine. The non-detection of hidden anti-nuclear antibodies by tissue sections in the absence of a splitting agent may be due to the formation of high molecular weight complexes between rheumatoid factors and anti-nuclear antibodies. These high molecular weight complexes containing anti-nuclear antibodies do not have access to tissue nuclear antigens, but can react directly with free nuclei. It is postulated that anti-nuclear antibodies may represent the early pathway of both rheumatoid arthritis and connective tissue diseases. The demonstration of hidden anti-nuclear antibodies in seropositive sera indicates that rheumatoid factors may have a protective effect. It may explain dissimilarities observed in the clinico-immunological profile of rheumatoid arthritis and systemic lupus erythematosus. The splitting effect of penicillamine observed in vitro may be similar in vivo. It can explain clinical improvement and immunological side effects observed in rheumatoid arthritis patients treated with this drug.

[Interactions between rheumatoid factors and antinuclear autoantibodies: their occurrence in a concealed state in rheumatoid disease (arthritis); effect of D-penicillamine]. / Interactions entre des facteurs rhumatoïdes et des autó-anticorps antinucléaires: fréquence de ceux-ci à l'état masqué au cours de la maladie (arthrite) rhumatoïde; action de la D-Pénicillamine.

In this paper it is demonstrated that IgM rheumatoid factor (RF) can inhibit the detection of antinuclear antibodies (ANA) by the classical rat liver cryostat section method. This "inhibition" of IgG-ANA by IgM-RF may be due to the formation of high molecular weight complexes. It would be the same in vivo. This masking effect can explain interesting similarities and dissimilarities observed in the clinico-immunological profiles of rheumatoid arthritis and systemic lupus erythematosus. D-Penicillamine can restore ANA activity by dissociating ANA-RF complexes. A new technique is described for the detection of these masked ANA.

Interactions between IgM antiglobulins and IgG antinuclear antibodies. Some aspects of D-penicillamine activities.

In this report we describe an in vitro masking action of IgM rheumatoid factor (IgM-RF) towards IgG antinuclear antibodies (IgG-ANA) which can be recovered by using D-penicillamine (DP) as an unmasking agent. The mechanism of this masking effect was elucidated by using smears of rat free nucleus as substrate, instead of the classical rat liver cryostat sections technique. It was postulated that the 'inhibition' of IgG-ANA by IgM-RF may be due to the formation of high molecular weight complexes (HMWC); this would be the same in vivo. Allowing the formation of HMWC which can be removed from the circulation, IgM-RF may have a protective effect by preventing or minimizing systemic lesions. Therefore, IgM-RF may be considered as a defence response against potentially noxious ANA or antigen-antibody complexes. Induced nephropathy and the high frequency of detection of ANA observed in rheumatoid arthritis (RA) patients treated by DP may be the result of the dissociating effect of this drug on HMWC, in which the activity of pre-existing ANA is hidden when rat liver sections are used for its detection.

[Variations in salivary immunoglobulins in Sjögren's syndrome (author's transl)]. / Variations des immunoglobulines salivaires au cours du syndrome de Gougerot-Sjögren.

A study of salivary immunoglobulins (IgA, IgG, IgM) was made in 74 subjects with or without Sjögren's syndrome. In the normal subjects, only IgA could be detected by classical immunodiffusion techniques. A correlation was sought between the presence of IgG and/or IgM in the saliva, and various clinical or objective clinical examinations, as well as with accessory salivary gland biopsy. This study indicates that in Sjögren's syndrome the detection of IgG and/or IgM is strongly correlated with sialography which is the surest examination used clinically. Thus the detection and estimation of salivary immunoglobulins is thus a simple and specific complementary examination in Sjögren's syndrome.

A new experimental model of systemic connective tissue disease in immunized rabbit.

In rabbit, the immumizations with whole Mycobacterium Tuberculosis injected intradermally and its crude cytoplasmic W.S.E. injected intraarticulary induce not only a self-perpetuating synovitis in the stimulated knee but also chronic systemic lesions of connective tissue (non-stimulated knee, both shoulders, aortic artery adventitial valvular endocardium, liver, kidneys and lungs). W.S.E. stimulates precipitating antibody synthesis, skin reaction and lymphoblastic transformation of lymphocytes in vitro. Furthermore, rabbits develop tuberculin skin test and lymphocytic response to P.P.D. The intensity of inflammation in stimulated knee is in direct ratio to intensities of humoral and cellular immunological responses. On the contrary, the incidence of systemic lesions is in inverse ratio to immune responses and intensity of inflammation in stimulated knee.

Salivary immunoglobulin determinations: their diagnostic value in Sjögren's syndrome.

Salivary immunoglobulins (IgA, IgG, IgM) determinations are performed on 74 patients with and without Sjögren's syndrome (SS). In normal subjects IgA is the only immunoglobulin detected in saliva by classical immunodiffusion methods. Correlations between the presence of IgG and/or IgM in saliva and other functional and objective clinical parameters and hsitological aspects of minor salivary glands are studied. In SS the prescence of IgG and/or IgM is in close relationship with "sialography index" which is the most reliable clinical investigation. These results point out that salivary immunoglobulins determinations constitute a simple and specific complementary test for SS diagnosis; they may allow the clinicians to observe the progress of the disease or to evaluate effectiveness of drugs.

Soluble complexes and antiglobulin factors detection by immunofluorescence and immunoadsorption in rheumatoid sera.

In this report is was demonstrated that in rheumatoid factors (RF) positive sera, 19 S IgM rheumatoid factor can form soluble complexes with different proteins (IgG, albumin) of sera. In these complexes the antiglobulin activity of IgM is not inhibited. When immunofluorescence and immunoadsorption procedures are used for the detection of antiglobulin activities of rheumatoid sera, the proteins which are bound to IgM rheumatoid factor, even if they are devoid of any antiglobulin character, may be revealed simultaneously with IgM. Moreover in some cases the detection of IgM may be hindered, while the linked proteins remain detectable. In these conditions, these complexes in RF positive sera may give false negative results for IgM rheumatoid factor, and may give rise to artefactual appearance of IgG and other proteins (albumin antiglobulin-like activities. This paper points out that before investigating IgG and IgA antiglobulin activities, IgM rheumatoid factor should be previously eliminated, for example by immunoadsorption.