Production and immunochemical characterization of Neisseria meningitidis group B antiserum for the diagnosis of purulent meningitis.

Unlike Neisseria meningitidis groups A, C, Y and W135, the group B capsular polysaccharide has been shown to be chemically and immunologically identical to the capsular polysaccharide of Escherichia coli K1. Both components are sialic acid homopolymers and are poorly immunogenic. Nevertheless, due to the high incidence of Neisseria meningitidis group B meningitis in the population of the State of São Paulo, preparing antiserum to this serogroup for diagnostic purposes has become a matter of high priority. Of the many immunization schemes proposed, intravenous inoculation of whole bacteria previously inactivated with formaldehyde and simultaneous intradermal inoculation with a mixture of the bacterial polysaccharide fraction and whole bacteria in complete Freund;s adjuvant have produced the best results. The antiserum was treated with immunoadsorbents prepared with aluminum chloride and protein and/or polysaccharide antigens from each of the following heterologous bacteria: Haemophilus influenzae type b, Streptococcus pneumoniae, Escherichia coli other than K1, and Staphylococcus aureus, in order to eliminate cross-reactivity. For quality control analysis, the antiserum was assessed by the immunodiffusion, counterimmunoelectrophoresis, dot-ELISA, and immuno-blot techniques against homologous antigens. Specificity was obtained after treating the antiserum with Haemophilus influenzae type b polysaccharide immunosorbent.

[Detection of Neisseria meningitidis group B antigens by MB-Dot-Elisa test in patients with meningitis]. / Detección de antígenos de Neisseria meningitidis del grupo B con la prueba MB-Dot-ELISA en pacientes con meningitis.

Infection with Neisseria meningitidis group B has been difficult to detect, partly because this bacterial group's polysaccharide is a weak immunogen. This article describes work carried out to test a new procedure (MB-Dot-ELISA) employing a high-titered horse antiserum for detection of N. meningitidis group B antigens. The study assayed cerebrospinal fluid samples from 585 subjects, 574 with suspected meningitis cases and 11 with neurologic disorders. The results of the assay indicated a sensitivity of 0.991 and a specificity of 0.826. These results were superior to those obtained with latex agglutination and in substantial agreement with the results of counterimmunoelectrophoresis and bacteriologic methods. Overall, the MB-Dot-ELISA was found to be sensitive, inexpensive, and suitable for public health laboratory investigations.

South American pemphigus foliaceus: electron microscopy and immunoelectron localization of bound immunoglobulin in the skin and oral mucosa.

Five skin and two oral biopsies from patients with South American pemphigus foliaceus (SAPF) were studied by electron and immunoelectron microscopy for the ultrastructural localization of bound immunoglobulin in epidermal and oral lesions. Electron microscopy showed the tonofilament-desmosome complex to be preserved in the various layers of the epidermis. Immunoglobulin was bound over the plasma membrane and permeated the desmosomal junctions both in the skin and oral mucosa, thus suggesting that pemphigus antibodies are attached to the glycocalyx. It appears that the initial injury in SAPF acantholysis involves the glycocalyx and that it might be caused by interaction with intercellular antibodies present in the patient's serum.

Chronic human schistosomiasis mansoni-schistosomal antigen, immunoglobulins and complement C3 detection in the liver.

The distribution of schistosomal antigen, immunoglobulins and complement C3 was studied by IF stain in 26 biopsies of human liver from 21 cases of hepatosplenic and five of intestinal schistosomiasis mansoni. Schistosomal antigen and immunoglobulins, chiefly of the IgG class and in a lesser intensity complement C3, were seen focally as scanty deposits in cells of the sinusoidal wall. They probably correspond to antigen-antibody insoluble large aggregates which are being removed by local cells of the mononuclear phagocyte system. Gamma globulin of the IgG class and antigens were also present in the granuloma around S. mansoni eggs and dead worms in the human liver. During the early phase of the granulomatous reaction the structure was not efficient enough to wall off completely the antigen, which is seen in cells at the center of the granuloma. As the granuloma matures, antigen demonstration becomes restricted to the miracidium, and immunoglobulins are observed mainly at the periphery. The kinetics of the granuloma formation with intralesional antibody presence promote a progressive antigen neutralization.

Detection of N. meningitidis group B antigens by MB-Dot-ELISA in patients with meningitis.

Infection with Neisseria meningitidis group B has been difficult to detect, partly because this bacterial group's polysaccharide is a weak immunogen. This article describes work carried out to test a new procedure (MB-Dot-ELISA) employing a high-titered horse antiserum for detection of N. meningitidis group B antigens. The study assayed cerebrospinal fluid samples from 585 subjects, 574 with suspected meningitis cases and 11 with neurologic disorders. The results of the assay indicated a sensitivity of 0.991 and a specificity of 0.826. These results were superior to those obtained with latex agglutination and in substantial agreement with the results of counterimmunoelectrophoresis and bacteriologic methods. Overall, the MB-Dot-ELISA was found to be sensitive, inexpensive, and suitable for public health laboratory investigations.

Production and immunochemical characterization of Neisseria meningitidis group B antiserum for the diagnosis of purulent meningitis

Unlike Neisseria meningitidis groups A, C, Y and W135, the group B capsular polysaccharide has been shown to be chemically and immunologically identical to the capsular polysaccharide of Escherichia coli K1. Both components are sialic acid homopolymers and are poorly immunogenic. Nevertheless, due to the high incidence of Neisseria meningitidis group B meningitis in the population of the State of São Paulo, preparing antiserum to this serogroup for diagnostic purposes has become a matter of high priority. Of the many immunization schemes proposed, intravenous inoculation of whole bacteria previously inactivated with formaldehyde and simultaneous intradermal inoculation with a mixture of the bacterial polysaccharide fraction and whole bacteria in complete Freund;s adjuvant have produced the best results. The antiserum was treated with immunoadsorbents prepared with aluminum chloride and protein and/or polysaccharide antigens from each of the following heterologous bacteria: Haemophilus influenzae type b, Streptococcus pneumoniae, Escherichia coli other than K1, and Staphylococcus aureus, in order to eliminate cross-reactivity. For quality control analysis, the antiserum was assessed by the immunodiffusion, counterimmunoelectrophoresis, dot-ELISA, and immuno-blot techniques against homologous antigens. Specificity was obtained after treating the antiserum with Haemophilus influenzae type b polysaccharide immunosorbent.