Does peripheral blood T-lymphocyte population distribution in sarcoidosis provide a prognostic clue?

In its pulmonary form, sarcoidosis generally resolves spontaneously, but it may lead to fibrosis of the lung. The clinical, radiological and functional tests, as well as activity markers such as the serum angiotensin converting enzyme, intrathoracic uptake of 67Gallium and the cytological data provided by bronchoalveolar lavage are only the expressions at any given time of a disease which is constantly progressing and only partly express its evolutive potential. The authors studied the distribution of T-lymphocyte subsets in the peripheral blood and from bronchoalveolar lavage. 32 patients were included in the study. They were suffering from acute or chronic sarcoidosis of the mediastinum and lungs and were divided into 2 groups according to clinical, radiological and pulmonary function criteria; Group A (n = 19) included regressive forms (minimum follow up 2 years) and group B (n = 13) the progressive untreated forms. Lymphopenia with a decrease in the percentage of CD3 cells was found in both groups. The percentage of CD4 cells is significantly lower in group B (28 +/- 11%) than in group A (45 +/- 8%) (p < 0.01) or in the control population (46 +/- 8%) (p < 0.01). The percentage of CD8 cells is higher in group B (30 +/- 8%) than in group A (18 +/- 6%). This results in a CD4/CD8 ratio which is significantly reduced in group B (1 +/- 0.5) when compared with group A (2.72 +/- 0.8) (p < 0.01) and the control group (2.17 +/- 0.8) (p < 0.01), the difference between group A and the controls being minimal.(ABSTRACT TRUNCATED AT 250 WORDS)

The relationship between CD5-expressing B lymphocytes and serologic abnormalities in rheumatoid arthritis patients and their relatives.

The influence of genetic factors on the expression of CD5+ B lymphocytes and their relationship to a broad spectrum of autoantibodies was investigated in a study of 12 patients with rheumatoid arthritis (RA) and 52 of their healthy first-degree relatives. The proportion of CD5+ B cells was significantly higher in RA patients (mean +/- SEM 23.9 +/- 2.7%) compared with that in their relatives (18.3 +/- 1.1%, P less than 0.05) and compared with that in a group of healthy control subjects (16.1 +/- 1.8%; P less than 0.05). Much more striking, however, were the high levels of CD5+ B cells found in the patients and their relatives in 5 of the families studied. Increases in total immunoglobulin levels and autoantibody levels were frequently observed in RA patients (approximately 20-40%) and their relatives (approximately 10-20%). Furthermore, a statistically significant correlation (P less than 0.01) between IgM rheumatoid factor and the percentage of B lymphocytes expressing CD5 was observed.

Epitopic analysis of HLA DR molecule expression on alloreactive T-cell clones.

The expression of HLA-DR epitopes, recognized by a set of anti-DR MoAbs clustered into four groups according to immunochemical studies, was analyzed at the surface of in vivo-sensitized alloreactive T-cell clones derived from a rejected kidney allograft and on the autologous B lymphoblastoid cell line (BLCL). Although no clear-cut differences were noted between T cells themselves and T cells vs BLCL in the relative expression of most of the DR epitopes studied (D1.12, BT2/9, VI.15C, 135, 206, 141, BM50), relative and absolute expression of one DR epitope (BMMag8) was strikingly higher on autologous BLCL than on T-cell clones. Moreover, the absolute expression of DR epitopes was heterogeneous among T cells. Such differences could not be explained either by T-cell activation level (assessed by IL2 receptor or transferrin receptor expressions) or by differences in time kinetics expression. In other tests, anti-DR MoAbs were assayed for their ability to block cytotoxic activity of several T-cell clones directed against DR allospecificities. Each T-cell clone showed distinct inhibition patterns. By such analysis, it was possible to define several groups of epitopes not always identical to those defined by immunochemical studies. Finally, analysis of the ability of MoAb to specifically block cytotoxicity mediated by 2 anti-DRw8 T-cell clones at the target level allowed precise epitopic study of polymorphic DRw8 determinants recognized by these cells, in this case borne by a single DR alpha/beta heterodimer.

[Evaluation of the activity of leukocytes during experimental periodontitis in the monkey]. / Evaluation de l'activité des leucocytes au cours d'une parodontite expérimentale chez le singe.

Polymorphonuclear leucocytes play an important role in the defense mechanisms and destruction of periodontal tissues. The leucocytes number and activity were compared during an induced periodontitis in 3 Cercopithecus cephus. On day 0, the 3 animals presented a localized marginal gingivitis. On 2 animals a severe marginal periodontitis was induced unilaterally: 42 days later leucocytes were sampled in the peripheral blood, in the gingival blood and in the gingival sulcus on the experimental and control sites as well as on the 2 hemijaws of the control animal. For each sample, a leucocyte count and a chemiluminescence test were done. Different results related to the disease severity were obtained with these tests for gingival blood and sulcular fluid.

[Anti-CD3 monoclonal antibodies. Characterization and function]. / Anticorps monoclonaux anti-CD3. Caractérisation et fonctions.

Four anti-T lymphocyte monoclonal antibodies were produced by the hybridoma technique. Percentages of miscellaneous labelled cell suspensions, as profiles of fluorescent histograms, showed a reactivity of these 4 reagents quite comparable to that observed with anti-CD3 monoclonal antibodies. Labelling summation studies, cell-sorting studies and blocking experiments gave similar results as gave other anti-CD3 reagents. Biochemical study and functional tests: modulation effects and mitogenic properties, argued for the belonging of these 4 antibodies to the CD3 cluster. The actual data concerning the CD3 molecule are briefly resumed to underline the interest of such antibodies in the understanding of T-lymphocyte activation mechanisms during the immune response.

[Anomalies of cellular immunity in Graves' disease treated by synthetic antithyroid drugs: effects on prognosis]. / Anomalies de l'immunité cellulaire dans la maladie de Basedow traitée par antithyroïdiens de synthèse: incidence sur le pronostic.

The involvement of humoral immunity mechanism in etiology of Graves' disease was observed for the first time in 1956. Twenty years later cellular auto-immune dysfunction was also described. To day, there is evidence for suppressor T cell deficiency and decrease in T suppressor lymphocytes in Graves' disease and cell mediated immunity might be one of the most important point in the pathogenesis of Graves disease. For the future, cellular auto-immune dysfunction might be used to predict the evolution of the disease and to choice the best therapeutic scheme.

[Chemiluminescence of phagocytic cells in chronic gingivitis and adult periodontitis]. / Chimioluminescence des cellules phagocytaires au cours des gingivites chroniques et des parodontites de l'adulte.

Measurement of chemiluminescence (CL) produced by phagocytic cells spontaneously or upon stimulation by phorbol myristate acetate (PMA) was performed in patients with gingivitis and adult periodontitis, and in control healthy subjects. The study was performed simultaneously on phagocytic cells obtained from peripheral blood and from gingival blood, and on crevicular leukocytes. An elevated CL production was obtained in quiescent and PMA-stimulated phagocytic cells from peripheral blood in patients with gingivitis or periodontitis compared to non-diseased controls. Chemiluminescence produced by unstimulated crevicular phagocytes was similar to that of peripheral blood phagocytes in normal subjects, but it was decreased in periodontitis. Upon PMA stimulation, the CL response of crevicular phagocytes remained low in the three groups of subjects.

[T lymphocyte subsets in alveolar lavage and peripheral blood in sarcoidosis and extrinsic allergic alveolitis]. / Etude des sous-populations lymphocytaires T dans le lavage alvéolaire et le sang périphérique dans la sarcoïdose et les alvéolites allergiques extrinsèques.

Accumulation of inflammatory and immune cells within lung parenchyma would constitute the initial step in producing the alveolar structural abnormalities. It is usually assumed that alveolitis, as assessed by broncho-alveolar lavage (BAL), represents a biological assessment of lung disease activity. The aim of this study, using monoclonal antibodies, is to characterize the T lymphocytes alveolitis in the lung and in peripheral blood in 3 well-defined populations: 1 degree) control subjects (n = 7); 2 degrees) patients with biopsy proven mediastino-pulmonary sarcoidosis (sarc) (n = 73), classified according to their clinical activity as active, inactive, chronic, and treated; 3 degrees) patients with extrinsic alveolar alveolitis (EAA) (n = 19). For the same BAL volume, the % of CD4+ cells and the CD4/CD8 ratio are increased in chronic and active sarc, contrasting with an increase in the % of CD8+ cells and a decrease in the CD4/CD8 ratio in the EAA. In absolute values, there are 2 times as many CD4+ cells and 5 times as many CD8+ cells in EAA than in sarcoidosis. In sarcoidosis, corticotherapy tends to normalize the CD4/CD8 ratio although the intensity of the lymphocytic alveolitis is not affected. In the peripheral blood, lymphopenia is observed only in the active form of sarc. in the CD4+ population, without any significant change in the CD4/CD8 ratio compared to the other groups. The number and distributions of BAL. T lymphocytes subsets may constitute a biological indicator for diagnostic orientation, but they do not distinguish sufficiently between the different groups of sarcoidosis to be of any prognostic value.

[Demonstration of new class I antigens in man]. / Mise en évidence de nouveaux antigènes de classe I chez l'homme.

The hypothesis of new class I antigens has been postulated in man, and several antigen systems have been proposed: HT (Gazit), TC (TCA, TCB) (Van Leeuwen). The present study describes a new class I antigenic marker system, expressed selectively on PHA-activated T lymphocytes and on lymphoblastoid B cell lines. These markers correlated at the cellular activation stage, have been called: human activation or HA markers. 7% of the sera from multiparous women present anti-HA antibodies. The definition of class I molecule (dimer 41K - 12K) has been established by a structural analysis of the molecule; the responsible gene, located on the 6th chromosome could be close to the HLA-A gene. The equivalence with the mouse Qa markers is postulated, but remains to be totally demonstrated.

A common epitope between HLA-B27, -B13 and -B37 alloantigens defined by a monoclonal antibody.

An anti-HLA-B27 monoclonal antibody produced by the hybridoma technique is described. This BD.7 reagent is a cytotoxic IgM antibody. Its reactivity was studied by lymphocytotoxicity tests, indirect immunofluorescence tests and biochemical analysis against an extensive panel of peripheral blood mononuclear cells. All HLA-B27 positive samples, either from normal subjects or from patients with Ankylosing Spondylitis, were recognized by this reagent. Moreover, a cross-reaction was observed with HLA-B13 cells, and a new unexpected reaction with all HLA-B37 cell suspensions. The interest of such a reagent is discussed.

Differentiation of thymocytes during human ontogeny: stage-specific DNA ligase in relation to terminal deoxynucleotidyl transferase, cell size and surface antigen.

The activities of two forms (7.5 and 5.5 S) of DNA ligase and of terminal deoxynucleotidyl transferase (TdT) have been studied in human thymocytes at different ages from 20 weeks pre-natal to 37 years after birth. Thymocytes have been selected on the basis of relative size and antigenicity (CD3, OKT3 immunofluorescence) with the cell sorter. For DNA ligases, three kinds of cells can be distinguished: (i) large antigenically negative cells of 20-week fetus, expressing only the 7.5 S enzyme; (ii) large antigenically positive cells without ligase activity; (iii) smaller antigenically positive cells, expressing only the 5.5 S enzyme. This last form of enzyme is found after birth. With respect to TdT expressed in OKT3- 5 micron cells and to OKT3+ thymocytes, it is observed that 5.5 S DNA ligase is found in a thymocyte population distinct from cells expressing TdT. Therefore, these results allow us to consider the 5.5 S DNA ligase activity as an additional functional marker for thymocyte maturation in humans.

Flow cytometric quantitative evaluation of phagocytosis by human mononuclear and polymorphonuclear cells using fluorescent nanoparticles.

The use of fluorescent polymethacrylic nanoparticles (0.3 micron) as a flow cytometric reagent in the quantitative evaluation of phagocytosis by human mononuclear and polymorphonuclear cells is described. The preparation of the nanoparticles, by emulsion copolymerization of methacrylic monomers, and their physicochemical properties are briefly summarized. Nanoparticles coupled with a fluorescent agent (ethidium bromide) were used in a flow cytometric assay to study opsonin-independent phagocytosis by human polymorphonuclear cells and by human monocytes. The phagocytosis of nanospheres by monocytes was determined by flow cytometry from the fluorescence distribution and ingestion was visualized by scanning and transmission electron microscopy. One possible application of the fluorescent nanoparticles is the simultaneous analysis of cell surface antigens and cell phagocytic activity.

New class I in man: serological and molecular characterization.

New class I antigens in linkage disequilibrium with HLA-A antigen are demonstrated in PHA T and EBV preferential target cells using human alloantisera. These new antigens are defined as class I antigens by immunoprecipitation of a 41-12 k dimer. The molecule is shown to be distinct from the HLA-A, -B, -C molecule and in particular from the A3 molecule as in sequential immunoprecipitations, the depletion of the HLA-A, -B, -C molecule or A3 molecule (44-12 k) has no effect on the new molecule (41-12 k). Being present on the PHA T cells and lymphoblast lines, these antigens are considered as new epitopes involved in the the cell activation process.

A monoclonal antibody (3A33) that reacts with a mouse-specific epitope of Mac-1 antigen.

The 3A33 monoclonal antibody, obtained by fusing rat immune lymphocytes with mouse plasmacytoma cells, was directed against mouse macrophages. Antibody 3A33, a rat IgG2a, reacted with macrophages from all the mouse strains tested, with mouse blood monocytes and with 56% of bone marrow cells, but not with T lymphocytes. It immunoprecipitated an antigen with alpha and beta subunits, found to be identical to Mac-1 antigen after cross-absorption experiments with M1/70 monoclonal antibody. The two antigenic determinants of the Mac-1 molecule identified by the 3A33 and M1/70 antibodies both displayed reduced expression on inflammatory macrophages and comparable resistance to trypsin digestion. The sites of the determinants on this molecule seemed close together judging from the ability of both the 3A33 and M1/70 antibodies to block C3bi receptor sites and compete for cell binding. However, unlike antibody M1/70, 3A33 never reacted with human cells bearing Mac-1 antigen. Therefore, two closely related epitopes of the Mac-1 molecule - one specific for mouse and one common to mouse and man, were recognized by these monoclonal antibodies.

Modulation of expression of mouse macrophage surface antigens by monoclonal antibodies.

Mouse macrophages from peritoneal cavity were exposed to monoclonal antibodies (MAbs) directed against cell surface antigens and the effect on antigen expression was investigated. The two Mabs used, 3A33 and 3A35, were produced by cell fusion between a mouse plasmacytoma and rat lymphocytes immunized against mouse macrophages. The binding of the MAbs to cell surface was measured by immunofluorescence and flow cytometry or by a radioimmunological technique. When injected i.p. the MAbs diminished the expression of the corresponding antigens but did not alter it when added to cultures of adherent macrophages. Antigenic modulation, however, could be produced in vitro either by inhibiting macrophage adherence during incubation with MAbs or by using a second antibody layer. MAb 3A33 (IgG2a) was more effective than 3A35 (IgM) in provoking modulation. The appearance of re-synthesized antigens on cell surface was not affected by macrophage adherence. The modulated antigens were found to internalize into cytoplasmic vacuoles.

Spleen cell populations in normal and tumor-bearing hamsters.

Spleen cell subpopulations from normal and tumor-bearing hamsters (TBH) were quantified using Petri dishes coated with specific antibodies, and by flow cytometric immunofluorescence analysis. The relative numbers of T cells (Thy + cells) decreased, by both methods, as a function of tumor growth, while the number of B cells (bearing surface Ig) increased. Cells without T or B markers (null cells) were more numerous in the spleen of TBH and a large number of them expressed the receptor for the Fc fragment of IgG. Splenic cells were also sorted according to their light-scattering properties, and electron microscopic analysis was performed on the sorted fractions. It showed the presence of secreting plasmocytes and activated macrophages in the spleen of TBH.

Anti-HLA-A2 and -A28 monoclonal antibody: production and study of the cross-reaction.

An anti-HLA-A2 and -A28 monoclonal antibody, XV.17, has been prepared by immunizing a Balb/c mouse with PBL. This XV.17 monoclonal antibody is a cytotoxic IgM. Its reactivity was tested by lymphocytotoxicity test and indirect immunofluorescence technique, in parallel with an alloantiserum ORA having the same anti-HLA-A2, -A28 reactivity pattern, against different panels. Family studies were undertaken. Absorptions-elutions and cytofluorometry experiments were performed to study the cross-reaction. The XV.17 monoclonal antibody is cytotoxic against all the HLA-A2 and -A28 tested cells, and is absorbed by HLA-Aw23 and -Aw24 cell suspensions.

Characterization and sorting of mouse cytotoxic macrophages by their light scattering properties.

The light scattering properties of mouse activated macrophages were analyzed by flow cytometry. Peritoneal adherent cells from B. abortus treated animals were found to segregate into two subpopulations as a function of their forward angle and 90 degrees angle light scatter. The cell subpopulations were separated by automatic sorting. The strongly scattering ones contained an elevated proportion of large volume and acid phosphatase rich cells. Their nonspecific cytotoxic activity against tumor cells was more important than that of weakly light scattering cells. Thus, flow cytometry might be helpful to characterize and isolate cytotoxic macrophage populations.

[Relation between cyclic AMP and phagocytosis in human monocytes]. / Relation entre AMP cyclique et phagocytose dans les monocytes humains.

The involvement of cyclic adenosine 3'-5' monophosphate (cAMP) in the regulation of human monocyte phagocytosis of staphylococcus aureus in vitro was demonstrated by assay of adenylate cyclase (AC) and cAMP phosphodiesterase (PDE). Phagocytosis was associated with diminished AC activity (p less than 0,05) and concurrently increased PDE activity (p less than 0,005). Transmission electron microscopy provided evidence that these changes coincide with peak phagocytic activity occuring 10-20 minutes after the start of phagocytosis.

A monoclonal antibody to the HLA-A3 alloantigen.

Monoclonal antibody production recognizing the HLA-A3 antigen is described. The XI-23 antibody reacted with all of the 89 cell suspensions carrying the HLA-A3 antigen (100% cytotoxicity) among a total of 191 suspensions tested. No extra-reactivity or cross-reactivity was observed, particularly with that of HLA-A11. This antibody can thus be considered as a good HLA-typing reagent.