Detection of rubella virus in amniotic fluid by electron microscopy.

We report in this paper the observation of rubella virus by electron microscopy in an amniotic fluid sample, collected from a pregnant woman with rubella infection. Virological investigations by inoculation of cell cultures with amniotic fluid and fetal blood remained negative, due probably to the presence of neutralizing antibodies in the samples. Electron microscopy is a rapid but weakly sensitive method to detect viruses in clinical specimens. However, this unusual observation would indicate that in some cases electron microscopy could be a useful technique to evidence a fetal rubella infection.

A study of beta 2-microglobulin skin deposits in dialyzed patients and healthy controls.

This study reports on beta 2-microglobulin (beta 2M) deposits in the skin of 12 uremic patients and three kidney transplant recipients compared with eight healthy controls. Uremic patients were treated by hemodialysis (HD), hemofiltration (HF), hemodiafiltration (HDF), or continuous ambulatory peritoneal dialysis (CAPD) for a period lasting from 1 to 19 years. Congo red staining of the skin was negative in patients and controls. However, immunofluorescent staining with an anti-beta 2-microglobulin monoclonal antibody was positive in the skin of all patients and of six of the eight controls. Beta 2M skin deposition is more intense in patients than in controls and increases with patient age and the duration of dialysis. A stron correlation is observed between the extent of skin beta 2M deposits and clinical manifestations due to beta 2M deposits in internal organs. However, no correlation is found between beta 2M skin deposits and sex or beta 2M serum levels.

Skin test reactions to a phenol-soluble antigen of Brucella abortus among veterinary students, Lyon, France, 1984.

In December 1984, 424 students at the National Veterinary College, Lyon, France, were skin-tested with a phenol-soluble antigen of Brucella abortus B19. Of all students, 2.6% had a positive intradermal reaction indicating previous contact with Brucella. Prevalence of positive reactions was significantly lower among students from the three first school years (less than 2%) compared with students in their last school year (5.9%). These results are discussed and compared to the prevalence of brucellosis among cattle. Specificity (94%) of the intradermal testing resembles that of serological testing but its sensitivity (75%) seems to be better.

[Hereditary deficiency of the 4th component of complement (C4) associated with a lupic syndrome]. / Déficit héréditaire du quatrième composant du complément (C4), associé à un syndrome lupique.

A 2 year-old girl presented with bacterial meningitis followed by a lupus erythematosus syndrome consisting of erythematous rash, Raynaud's phenomenon and mutilating cicatricia atrophy; high speckled antinuclear antibodies and anti-Sm and anti-Ro antibodies titers; a selective absence of C4; moderate mesangial proliferation with IgM and C1q mesangial deposits. Study of the family revealed a linkage of C4 deficiency with the HLA A1 B17/BfF haplotype.

Two sensitive assays for the detection of anti-Brucella antibodies.

A laser-nephelometric assay (LNA), applied to an adapted Wright's agglutination technique and an immunoenzymatic assay (ELISA) on tyndallised and sonicated Brucella are described. Both lead to an increased sensitivity of previous techniques for the detection of anti-Brucella antibodies. This was assessed in a) Brucellosis patients with either Wright and/or IF positive reactions (sero-positive) or Wright and IF negative reactions (sero-negative, ancient brucellosis). Positive results were obtained in all cases with a highly significant increase of the titers observed in "sero-positive" patients. b) Normal subjects with no previous history and no biological stigmata of brucellosis: most sera gave negative results - a very low incidence of weakly positive results were however evidenced. c) Subjects vaccinated with fraction PI: positive results were observed in all cases. They were much earlier detected with LNA or ELISA than with Wright's or IF tests and the ELISA or LNA titers of late "sero-positive" sera were 3 to 36 times higher than Wright's titers. d) Supernatants of anti-Brucella hydridomas: LNA and ELISA were both successfully applied to the screening of such supernatants.

[Membranous glomerulonephritis in two brothers associated in one with tubulo-interstitial disease, Fanconi syndrome and anti-TBM antibodies (author's transl)]. / Glomérulonéphrite extra-membraneuse chez deux frères associée dans un cas à une néphropathie tubulo-interstitielle avec syndrome de Fanconi et anticorps antimembrane basale tubulaire.

Anatomical and immunological studies were performed in two brothers with membranous glomerulonephritis. The older child presented with renal failure, Fanconi syndrome and anti-TBM antibody in his plasma. Renal biopsy revealed severe tubulo-interstitial disease with membranous glomerulonephritis. Because of rapidly progressive renal insufficiency the patient was started on hemodialysis. A cadaver renal allotransplantation was performed without success. Renal transplant biopsy showed severe lesions of vascular rejection without recurrence of the primary disease. The younger child was examined at six months for a nephrotic syndrome with mild renal insufficiency, and died at nine months. Anti TBM antibodies were not detected in his serum. The patient's mother presented with asymptomatic proteinuria, and anti TBM antibodies in her plasma. In the same family the uncle died at 3 months with a steroid resistant nephrotic syndrome. The relationships between the familial membranous glomerulonephritis and the tubulo interstitial disease are discussed.

[Extramembranous glomerulonephritis following hepatitis-B. Simultaneous disappearance of serum virus antigen and glomerular lesions (author's transl)]. / Glomérulopathie extra-membraneuse après hépatite à virus B. Disparition parallèle de l'antigénémie virale et de l'atteinte glomérulaire.

A 24-year old female patient developed extramembranous glomerulonephritis 8 months after acute hepatitis-B with virus antigen persisting in the serum. Two years later, the glomerular lesions were found to be clinically and histologically cured, with parallel disappearance of the antigen from the serum. Although the presence of HBsAg in the glomeruli could not be demonstrated, it is likely from the sequence of events that hepatitis-B virus was responsible for the renal disease.

Studies on the immune protection to murine experimental brucellosis conferred by Brucella fractions. I. Positive role of immune serum.

Mouse inoculation with three different phenol-insoluble fractions extracted from Brucella melitensis (fractions 'PI', '4A' and '5') induces an acceleration of the blood clearance of i.v. inoculated live Brucella and a diminution of the rate of multiplication of the injected bacteria in the spleen. Preincubation of the challenge inoculum in immune serum or i.p. injections of immune serum confer a good specific protection to non-immunized hosts. The results observed with fractionated sera suggest that, not only antibodies, but also other serum constituents may participate in the protective activity of immune sera. This discussed in terms of the respective importance of humoral and cellular immunity to Brucella and of the choice of the best preparations for human or animal vaccination.

[Recurrent hematuria in children]. / Hématuries récidivantes de l'enfant.

Twenty-three children with recurrent macroscopic hematuria have been observed. Six children presented with glomerular disorder with mesangial deposits of IgG and IgA (Berger's disease). The hematuria episodes occurred in the 48 hours following an infection, more often respiratory. Renal functions were not impaired. On light microscopy, glomerular lesions were minimal, or were lesions of focal and segmental glomerulonephritis. Ten children seemed to have hereditary nephritis. In 5 cases, nephritis was accompanied by deafness (Alport's syndrome). In 5, nephritis was hereditary but isolated. The electron microscopy examination of basement membranes showed lesions consistent with Alport's syndrome. In cases of hereditary nephritis without deafness, basement membranes were less impaired. In 7 children, no specific etiology was found.

Stimulating activity of Brucella fractions in a human lymphocyte transformation test. Correlation with humoral and cellular immunity.

'PI' a phenol-extracted fraction from Brucella melitensis has been used for years in our laboratory for the vaccination of laboratory and agricultural workers considered 'at risk'. In the present work it has been used in vitro, together with another phenol-extracted and peptidoglycan containing fraction (4A) in a lymphocyte transformation test. Correlation between stimulation index, humoral antibodies to Brucella and the cutaneous response to PI have been studied in subjects vaccinated with the PI fraction, in Brucellosis patients and normal controls.

[Membranoproliferative glomerulonephritis and mycoplasma infection]. / Glomérulonéphrite membrano-proliférative et maladie mycoplasmique

Two children presented with an acute mycoplasma infection associated with a significant increase of antistreptolysin level. A severe nephropathy occurred, rapidly resulting in renal failure with histologic lesions of membrano-proliferative glomerulonephritis. Both patients had persisting low complement levels with low C3 and normal C4. The relationship between mycoplasma and streptococcal infections and the abnormality of complement and the renal disease is discussed.

Immunization by an insoluble fraction extracted from Brucella melitensis: immunological and chemical characterization of the active substances.

A peptidoglycan-containing fraction called fraction P.I. (phenol insoluble), extracted from Brucella melitensis and previously described by some of us, had immunogenic and protective properties and did not produce any allergic reactions. Since it is well known that bacterial peptidoglycans studied so far have immunoadjuvant properties, the isolation of the active factor(s) of Brucella was undertaken. By successive enzymatic and chemical treatments, a new, much more purified fraction, called "4A" (approximately 5% of fraction P.I.), is obtained, retaining the same properties as P.I. and giving better protection against infection by Brucella. Immunogenicity, immunoadjuvant activity, allergizing capacity, and specific and nonspecific protective effects of fractions P.I. and 4A are compared. Chemically, fraction 4A is constituted by a lipoprotein covalently linked to peptidoglycan and by a few (lipo)proteins that could be solubilized by hot sodium dodecyl sulfate. Intrinsic properties of peptidoglycan could not be studied, but it does not seem to be essential for the activity. In conclusion, fractions P.I. and 4A are not agglutinogenic and, since fraction 4A induces better protection against infection by Brucella, it could advantageously replace fraction P.I. as a vaccine for humans.

[Lymphoblastic transformation by fractions of Brucella in patients having had brucellosis and subjects vaccinated by the vaccine PI]. / Transormation lymphoblastique par des fractions de Brucella chez des malades atteints de brucellose et des sujets vaccinés par le vaccin P.I

Two fractions of brucella, examined elsewhere for their vaccinal properties (P.I. and 4A) have been used in lymphoblastic stimulation tests with lymphocytes of subjects infected by brucellosis and subjects vaccinated with P.I. This investigation attempted to show the importance of cellular immunity in brucellosis and to deduce a valid protection test. The two fractions do not possess non-specific mitogenic activity for human lymphocytes, but they stimulate specifically the sensitized lymphocytes. Subjects formerly or recently infected with brucellosis present constantly positive tests with sometimes a very high stimulation index. Vaccinated subjects have a lower stimulation index which is, however, significant in practically each case. The P.I. fraction gives the most constant results in the two categories of subjects; 4A appears to be much less active. The divergence between the protection rates conferred by 4A to animals and the weak activity of this same fraction in stimulation tests of human lymphocytes raises, on the one hand, the problem of the relation between protection and cellular immunity and, on the other hand, the utilization of animal models for the investigation of human vaccines.

[Chemical fractionation of Brucella melitensis and the immunizing properties of a new fraction]. / Fractionnement chimique de Brucella melitensis et propriétés immunisantes d'une nouvelle fraction

A complex phenol-insoluble fraction (fraction P.I.) extracted from Brucella melitensis presents immunizing properties already described by the authors. This fraction contains peptidoglycane whose immuno-adjuvant properties are known in numerous bacterial species. More precise fractionation was carried out to determine if the properties observed after injection of P.I. are in fact specific or on the contrary due to peptidoglycane action. By chemical and enzymatic treatments, 90% of the P.I. are eliminated, leaving a 4A fraction having the same properties as P.I. and having the advantage of providing better protection. This protection specific to genus Brucella and slow in appearing, is not due to peptidoglycane. The 4A fraction consists essentially of peptidoglycane linked covalently with a lipoprotein and proteins of weak molecular weight. All attempts at more precise fractionation have until now led to the loss of biological activity.