Inhibition of mixed lymphocyte reactions in humans after immunization with tetanus toxoid.

Immunization of humans with tetanus toxoid (TT) results in an inhibition of the reactivity of peripheral lymphocytes to TT as well as to one-way mixed lymphocyte reaction. The strongest inhibition was observed in cells taken 3 or 7 days after immunization. These reactivities reappeared on the 14th day and reached their initial levels by the 21st day after immunization. The role of the various cell types was studied by culturing separately purified T or B lymphocytes taken on day 7 after immunization. B cells alone are not stimulated by TT or allogeneic cells. Purified T cells are stimulated much more than the initial total cell population. When B cells were mixed and cultured along with T cells, a suppressor effect upon stimulation appeared which reduced 3H-thymidine incorporation to the levels of the initial population. Treatment with 5-bromodeoxyuridine followed by exposure to light of cells stimulated with TT abrogated their ability to respond to subsequent stimulation with TT; whereas the responsiveness of treated cells to allogeneic lymphocytes was not significantly affected and vice versa. These results suggest that each stimulator activated a separate population of T cells but that in vivo immunization provoked the development of both a specific and nonspecific suppressor activity. Most probably this suppressor activity was brought about by adherent, surface Ig-bearing (B) cells.

Idiotype restriction in human autoantibodies to DNA in systemic lupus erythematosus.

Antibodies (Ab) to double stranded DNA (dsDNA) were immunoaffinity-purified from the serum of patient TOF with active systemic lupus erythematosus (SLE). Anti-idiotypic Ab to TOF anti-DNA were raised in rabbit. They were shown to recognize TOF F(ab')2 fragments, but they did not interact with human Ab of other specificities or with TOF IgG depleted of anti-DNA activity. In addition, their binding to TOF idiotype was specifically inhibited by DNA molecules. These anti-idiotype Ab therefore recognized idiotopes associated with a variable region of TOF anti-DNA-combining site. Thirty-one purified IgG anti-DNA preparations from unrelated SLE sera were able to inhibit this idiotype-anti-idiotype reaction by up to 90%. However, there was no linear correlation between anti-dsDNA Ab levels and idiotype-blocking capacity. These findings suggest that, in SLE, the idiotypic repertoire of autoAb to dsDNA is more restricted than thought previously.

Cytotoxicity reactions against target cells infected with rabies virus.

Some aspects of the cytotoxicity reactions were studied in the rabies system. The antibody-dependent complement cytotoxicity (ADC), the cellular cytotoxicity (CC), and the antibody-dependent cellular cytotoxicity (ADCC) are shown, being the cytotoxic effect as evidenced by the 51Cr released from the cells infected with the Pasteur strain of rabies virus. Some parameters such as time of cellular infection, the amount of infected cells, the concentration of complement, and the incubation time of the ADC reaction, which help to increase the performance of this reaction, are discussed. The detection and the level of the cellular response against the Pasteur strain of rabies virus in immunized mice is shown. Evidence is presented that in the ADCC test, specific human antibodies and non-immune human lymphoid cells are able to mediate in vitro lysis of cells infected with rabies virus. A comparison of the ADCC test with serum neutralization and immunoenzymatic tests is shown.

[Ciliary activity of cells of the fallopian tubes (apropos of the sequellae of salpingitis)]. / Etude de l'activité ciliaire des cellules de la trompe de Fallope (à propos des séquelles de salpingite).

A study was made of ciliary movement in tubal epithelium using microphotooscillography on the tubes of 91 patients who were operated on for tubal, ovarian or uterine pathology. Three groups were classified: 31 with healthy tubes, 47 with tubes that showed the consequences of salpingitis. These patients had been operated on for sterility after much treatment with antibiotics and corticoids. 13 sets of tubes from pregnant women (6 cases had intra-uterine pregnancies and underwent tubal ligation after Caesarean section or after a termination of pregnancy) and 7 cases had extra-uterine pregnancies with salpingectomy. This study has made it possible for us to make several observations: The ciliary activity was not altered by the time in the menstrual cycle. When the tubes were healthy tubal activity was hardly changed when a fibroid was present, nor when an ovarian cyst or endometriosis were present (550 beats a minute). In pregnant women ciliary activity is maintained until the pregnancy becomes intra-uterine. It is always abolished in cases of extra-uterine pregnancy, even if this is at some distance from the site of inflammatory reaction caused by implantation of the oocyte. In cases following salpingitis, ciliary activity in the tubes is often altered and sometimes nil. It therefore seems that if one excludes anatomical lesions such as tubal blocks and ampullary phimosis, salpingitis which is the cause of sterility changes tubal physiology greatly. This is true even at some distance from the site of the infective process as is shown by changes in ciliary activity.

Antibodies to heterogeneous nuclear ribonucleoproteins in sera from patients with rheumatic autoimmune diseases.

Ribonucleoprotein (RNP) particles sedimenting at 40 S in sucrose gradients were prepared from calf thymus nuclei. They were identified as heterogeneous nuclear RNP (hnRNP) on the basis of size, electron microscopic examination, buoyant density, and protein electrophoretic patterns. Sera from patients with systemic lupus erythematosus, rheumatoid arthritis, and mixed connective tissue disease were found to interact with hnRNP by counter-immunoelectrophoresis and enzyme-linked immunosorbent assay (ELISA). Small nuclear RNP (snRNP) were purified by immunoaffinity using a monoclonal anti-snRNP antibody immobilized on Sepharose beads. Inhibition of the ELISA assay for snRNP with anti-hnRNP Fab fragments and cross-over experiments revealed that the autoantibodies detected in human sera recognize common epitopes present on snRNP and hnRNP.

Expression of anti-idiotypic clones against auto-anti-DNA antibodies in normal individuals.

A serum pool from 280 blood donors and individual samples from blood donors were assayed for anti-idiotypic activity to auto-anti-DNA antibodies by competitive radioimmunoassays. We found that serum from several normal blood donors inhibited the binding activity of anti-DNA antibodies affinity purified from the sera of patients with systemic lupus erythematosus. This inhibition was due to immunoglobulin molecules but was not due to rheumatoid factor activity. Antiallotypic antibodies were not responsible for the anti-anti-DNA activity detected. Because this inhibition was blocked by DNA molecules, the observed reactivity was probably caused by idiotype-anti-idiotype interactions. These results provide evidence that anti-idiotype antibodies against anti-DNA autoantibodies are present in certain normal human sera. Anti-anti-DNA antibodies could play a role in the regulation of autoimmunity to DNA.

Idiotypic/antiidiotypic interactions in systemic lupus erythematosus: demonstration of oscillating levels of anti-DNA autoantibodies and reciprocal antiidiotypic activity in a single patient.

This work was designed to investigate the occurrence and evolution of autoantiidiotypic antibodies to double-stranded DNA in systemic lupus erythematosus (SLE). Serum samples from a patient with SLE who was submitted to repeated therapeutic plasmapheresis for 57 weeks were examined for the presence of autoantiidiotypic antibodies to DNA. Anti-DNA antibodies were affinity-purified from a serum sample obtained at the beginning of the observation period. Anti-anti-DNA activity, directed toward structures within the binding site of anti-DNA autoantibodies but not antibodies of another specificity, was detected in the serum of the patient. In addition, the decrease in the levels of anti-DNA antibodies took place concomitantly with a reciprocal increase in the autologous anti-idiotype. These results indicate that anti-DNA and anti-anti-DNA antibodies coexist in the serum of this patient, and suggest that antiidiotypic interactions may play a role in the modulation of idiotypic expression in SLE.

Idiotypy of anti-Rh antibodies.

Anti-id sera to Rh antibodies were produced by injecting rabbits with purified Rh antibodies. These sera were shown to agglutinate O Rh+ RBC coated by the immunizing antibody and--in some cases--by other anti-D antibodies. Id and cross-reactive id were shown to be located in the antigen-binding and in the non-antigen binding regions of Rh antibodies. An unique example of evolution of idiotypic specificities on human antibodies has been reported. Lastly, we have demonstrated by rosette assay, presence on some PBL of receptors for Fab'2 anti-Rh coating O Rh+ red cells. Rosettes could not be obtained with lymphocytes of a donor and Fab'2 anti-Rh of another individual. Rosettes appeared at a period of time in which the amount of antibody was decreasing.

[Systemic lupus erythematosus and anti-native desoxyribonucleic acid antibodies: clinical and laboratory comparison of the results given by two methods of detection (author's transl)]. / Anticorps anti-acide désoxyribonucléique natif au cours du lupus érythémateux disséminé. Comparaison clinique et biologique des résultats de deux méthodes de recherche.

Simultaneous application of two methods for the detection of anti-native DNA antibodies (indirect immunofluorescence on DNA fibres and Farr's radioimmunological method with native DNA labelled C14) to 53 serum samples from 32 patients suffering from systemic lupus erythematosus (SLE) indicated a certain number of discordances between the results obtained. Correlations between clinical and laboratory findings were found, such that certains types of anti-native DNA antibodies (n) may be specific to certain clinical manifestations of SLE: a good correlation was seen between She fixation of anti-C14 nDNA and the titre of IF anti-nDNA in the group of SLE with renal involvement, but was not found in the group of SLE with central nervous system involvement. There was no parallel between the course of renal involvement and anti-nDNA antibody titres, without treatment being directly responsible. The significance of the presence or absence of anti-DNA antibodies with the two methods of detection used is not of any single significance. Several types of anti-nDNA antibodies are present in a given specimen of lupus serum. Certain anti-nDNA antibodies are not detected or are at the limit of a positive result in the presence of neurological manifestations. From a practical standpoint, the use of several laboratory tests is desirable for the possible investigation of SLE.

IgG-subclass expression of anti-DNA and anti-ribonucleoprotein autoantibodies in human malaria.

To understand further the autoimmune phenomena associated with human malaria, we examined the IgG-subclass expression of antibodies to DNA and to ribonucleoproteins (RNP) in the serum of 99 patients with acute malaria. Of the sera, 22% were positive for single-stranded DNA (ssDNA), 18% for double stranded DNA (dsDNA) and 32% for RNP. Using a set of human IgG-subclass-specific murine monoclonal antibodies, we found that autoantibodies to dsDNA were predominantly expressed in the IgG1 subclass. In contrast, anti-ssDNA antibodies were more evenly distributed among the three other isotypes. Antibodies to RNP were essentially of the IgG1 and IgG2 isotypes. However, there was no correlation between these restricted IgG-subclass in the sera. These results are discussed in the context of previous findings of isotype expression of these autoantibodies in patients with the autoimmune disease systemic lupus erythematosus.

Evaluation of auto-antibodies in chronic mucocutaneous candidiasis without endocrinopathy.

Six patients with chronic mucocutaneous candidiasis (CMCC) were investigated for the presence of auto-antibodies during the course of the infection. Sera were tested for antibodies to native DNA (dsDNA) and denatured DNA (ssDNA), mitochondrial and microsomal antigens, smooth muscle, gastric parietal cells, basal membrane and skin intercellular substance, parathyroid glands, thyroglobulin and microsomal antigen, immunoglobulins and for anti-nuclear antibodies. Auto-antibodies were detected by radioimmunoassay, immunofluorescence, hemagglutination and other routine methods. Tests were performed at the end of the observation period, with the same batches of antigens and at the same time for all patients. Organ-specific antibodies (gastric parietal cells and intercellular substance) were found at low titers in five patients. Anti-smooth muscle antibodies were increased in two patients. In four patients antibodies to ssDNA were elevated. Moreover high titers of anti-ssDNA antibodies correlated well with disease activity after treatment with Ketoconazole in four tested patients. The possibility that C. albicans infection may induce auto-antibodies should be considered in assessing their disease activity significance in other chronic infected patients. The mechanisms of appearance of auto-antibodies and their immunopathological significance in CMCC are discussed.

IgG subclass distribution of autoantibodies to DNA and to nuclear ribonucleoproteins in autoimmune diseases.

Fifty-seven serum samples positive for antibodies to double-stranded DNA (dsDNA), single-stranded DNA (ssDNA) or small nuclear ribonucleoproteins (snRNP) selected from patients with systemic lupus erythematosus and mixed connective tissue disease, were examined for the IgG subclass distribution of these autoantibodies. It was shown that antibodies to dsDNA were relatively restricted to IgG1 and IgG3 subclasses whilst antibodies to ssDNA were equally distributed throughout the four subclasses. Antibodies to snRNP were essentially restricted to the IgG2 isotype. These isotype distribution patterns contrasted with that observed for total serum IgG.

Mast cell sensitizing antibodies in myasthenia gravis.

Mast Cells Sensitizing Antibodies (MCSAb) were detected in patients with Myasthenia Gravis (MGr), by the Indirect Mast Cell Degranulation (IMCD). In most of the examined sera a correlation could be established between MCSAb and fluorescent antibodies to muscle and to thymus. The MCSAb were specific for muscle extract. They were resistant to prolonged heating and their reactivity in the IMCD test was not complement depending. Presence of MCSAb in sera of patients with MGr evidences the auto-allergic features of this disease.

[Experimental polymyositis induced by the injection of acetylcholine receptors. Study of the lesions in rats and guinea pigs]. / Polymyosite expérimentale provoquée par l'injection de récepteurs d'acétylcholine. Etude des lésions chez le rat et chez le cobaye

The injection of acetylcholine receptors, from the Electrophorus electricus induces in Guinea Pigs and Rats an experimental disease, which is characterized by the appearance of antibodies to sarcolemma and by massive histological alterations of the degenerative type, similar to those observed in clinical polymyositis.

Mast cell sensitizing antibodies in experimental myositis.

Mast cell sensitizing antibodies, direct mast cell degranulation and positive delayed skin tests were detected in guinea pigs and rats during experimental paralysis provoked by inoculation with acetylcholine receptors from the Electrophorus electricus. The animals also developed fluorescent antibodies to the sarcolemma, and massive myositis with histological changes in muscles similar to those characterizing clinical polymyositis.