Production of monoclonal antibodies anti-Taenia crassiceps cysticerci with cross-reactivity with Taenia solium antigens.

We describe the production of the potential monoclonal antibodies (MoAbs) using BALB/c mice immunized with vesicular fluid (VF)-Tcra (T. crassiceps) antigen. Immune sera presented anti-VF-Tcra (<20kD) IgG and IgM antibodies with cross-reactivity with T. solium (Tso) antigen (8-12, 14, and 18 kD). After cell fusion, we selected 33 anti-Tcra and anti-Tso reactive IgM-clones and 53 anti-Tcra specific IgG-clones, 5 of them also recognizing Tso antigens. Two clones identified the 8-14 and 18kD peptides of VF-Tcra.

Detection of Neisseria meningitidis in cerebrospinal fluid samples from suspicious cases of meningococcal meningitis using polymerase chain reaction and counterimmunoelectrophoresis.

Rapid diagnosis of meningococcal disease followed by an early treatment is essential. However, blood or cerebrospinal fluid cultures may not be successful because antibiotic treatment is often started before proper specimens are collected and because bacteria may die during transportation to the laboratory. Improvements in antibiotic therapy for specific microorganisms will require the use of more than one method for immunodiagnosis. In this study a collection of cerebrospinal fluid samples from Brazilian patients was analyzed. Gram stains, culture, counterimmunoelectrophoresis and clinical evaluations for meningococcal diseases were available. The sensitivity of nested PCR (nPCR) was 73% for cerebrospinal fluid of clinically suspected cases, whereas both sensitivity and specificity were 100% when subtypes of Brazilian epidemic strains (P1.7, P1.9 and P1.15) isolated from the samples were used.

Detection of Neisseria meningitidis in cerebrospinal fluid samples from suspicious cases of meningococcal meningitis using polymerase chain reaction and counterimmunoelectrophoresis

Rapid diagnosis of meningococcal disease followed by an early treatment is essential. However, blood or cerebrospinal fluid cultures may not be successful because antibiotic treatment is often started before proper specimens are collected and because bacteria may die during transportation to the laboratory. Improvements in antibiotic therapy for specific microorganisms will require the use of more than one method for immunodiagnosis. In this study a collection of cerebrospinal fluid samples from Brazilian patients was analyzed. Gram stains, culture, counterimmunoelectrophoresis and clinical evaluations for meningococcal diseases were available. The sensitivity of nested PCR (nPCR) was 73 for cerebrospinal fluid of clinically suspected cases, whereas both sensitivity and specificity were 100 when subtypes of Brazilian epidemic strains (P1.7, P1.9 and P1.15) isolated from the samples were used.(AU)

Detection of Neisseria meningitidis in cerebrospinal fluid samples from suspicious cases of meningococcal meningitis using polymerase chain reaction and counterimmunoelectrophoresis

Rapid diagnosis of meningococcal disease followed by an early treatment is essential. However, blood or cerebrospinal fluid cultures may not be successful because antibiotic treatment is often started before proper specimens are collected and because bacteria may die during transportation to the laboratory. Improvements in antibiotic therapy for specific microorganisms will require the use of more than one method for immunodiagnosis. In this study a collection of cerebrospinal fluid samples from Brazilian patients was analyzed. Gram stains, culture, counterimmunoelectrophoresis and clinical evaluations for meningococcal diseases were available. The sensitivity of nested PCR (nPCR) was 73 for cerebrospinal fluid of clinically suspected cases, whereas both sensitivity and specificity were 100 when subtypes of Brazilian epidemic strains (P1.7, P1.9 and P1.15) isolated from the samples were used.

Cross-reactivity of anti-Taenia crassiceps cysticerci immune antibodies with Taenia solium antigens.

A comparative study of antibody production was carried out using BALB/c mice immunized with 20 or 50microg vesicular fluid (VF)-Tcra (Taenia crassiceps) antigens, and gel of <30kD or eluate from <30kD peptides. Good IgM, IgA and IgG levels were detected by ELISA-Tcra and the antibodies presented reactivity with the <20kD peptides when tested by immunoblotting-Tcra. The antibodies from animals immunized with 20 and 50microg presented high anti-Tso cross-reactivity in ELISA (IgG>>IgM and IgA). All groups presented IgG antibodies identifying the 12kD Tso-peptide.

Production and characterization of a new monoclonal antibody against Neisseria meningitidis: study of the cross-reactivity with different bacterial genera.

We have generated a hybridoma cell line which produces an 8C7Br1 clone of the IgM antibody isotype. It recognizes the 50-, 65-, and 60-kDa antigens and is reactive with strains of N. meningitidis in the 98% of local Neisseria genera by Dot-ELISA assays. Two percent of the strains of N. meningitidis B do not present reactivity with the 8C7Br1 monoclonal antibody (MAb). The antibody reacted against N. meningitidis of serogroups A, B, C, X, Y, Z, and different serotypes and subtypes of N. meningitidis B and C by means of Dot-ELISA and Immunoblot. It cross-reacted with Neisseria gonorrhoeae, Neisseria lactamica, Haemophilus influenzae type b, Escherichia coli, Salmonella typhimurium, Salmonella typhi, Shigella flexneri, Bordetella pertussis, and Bacillus subtilis. The 8C7Br1 MAb reacted with the 65-kDa protein present in the prototype meningococcal strains B:16:B6(B2a:P1.5.2) and 2996 (B2b:P1.5.2). In H. influenzae type b, E. coli and B. subtilis, the MAb recognized the protein of 60, 65, and 70 kDa, respectively. FACS analysis showed that 8C7Brl MAb could recognize the 50-kDa protein on the surface of N. meningitidis homologous (B:4:P1.9) strain. These results, together with the bactericidal activity of 8C7Br1, and an experiment of passive protection in mice, demonstrated the potential importance of the cross-reactive protein as a candidate antigen for N. meningitidis B vaccine composition.

Detection of Neisseria meningitidis in cerebrospinal fluid samples from suspicious cases of meningococcal meningitis using polymerase chain reaction and counterimmunoelectrophoresis.

Rapid diagnosis of meningococcal disease followed by an early treatment is essential. However, blood or cerebrospinal fluid cultures may not be successful because antibiotic treatment is often started before proper specimens are collected and because bacteria may die during transportation to the laboratory. Improvements in antibiotic therapy for specific microorganisms will require the use of more than one method for immunodiagnosis. In this study a collection of cerebrospinal fluid samples from Brazilian patients was analyzed. Gram stains, culture, counterimmunoelectrophoresis and clinical evaluations for meningococcal diseases were available. The sensitivity of nested PCR (nPCR) was 73

for cerebrospinal fluid of clinically suspected cases, whereas both sensitivity and specificity were 100

when subtypes of Brazilian epidemic strains (P1.7, P1.9 and P1.15) isolated from the samples were used.

The use of filter paper monoclonal antibodies in a Dot-blot test for typing Neisseria meningitidis B.

A simple method for the collection, preservation, shipment, and testing of minute amounts of dried monoclonal antibodies for typing Neisseria meningitidis B is described. The monoclonal antibodies collected on filter paper were extracted in PBS and evaluated by Dot-blot employing whole cells of N. meningitidis B as antigen. The dried filter paper with monoclonal antibodies could be stored at room temperature for as long as 30 days without detectable changes in antibody response when used for typing outer membrane antigens of N. meningitidis B.

The use of filter paper monoclonal antibodies in a Dot-blot test for typing Neisseria meningitidis B

A simple method for the collection, preservation, shipment, and testing of minute amounts of dried monoclonal antibodies for typing Neisseria meningitidis B is described. The monoclonal antibodies collected on filter paper were extracted in PBS and evaluated by Dot-blot employing whole cells of N. meningitidis B as antigen. The dried filter paper with monoclonal antibodies could be stored at room temperature for as long as 30 days without detectable changes in antibody response when used for typing outer membrane antigens of N. meningitidis B

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.

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.

Trypanosoma cruzi: studies on the reactivity of antibodies bound to the surface of blood forms at the early phase of infection.

The specificity and reactivity of antibodies bound to the surface of Trypanosoma cruzi blood forms at the very early acute phase of murine infection was investigated. Surface-bound antibodies of the IgG and IgM isotypes were recovered from blood forms upon incubation at 37 degrees C. The eluted antibodies immunoprecipitated several trypomastigote surface polypeptides from 80 to 100 kDa. In contrast, for epimastigotes a very faint reactivity was detected only for antigens of 50 and 95 kDa. The shed antibodies promoted in vitro complement-mediated lysis of live blood forms and reacted with fixed trypomastigotes by immunofluorescence. Thus, blood forms are already coated with active trypomastigote-specific antibodies with a potential role in the host defense, although the low levels of serum antibodies have prevented the demonstration of humoral protection at the early stages of infection.

Trypanosoma cruzi: serum antibody reactivity to the parasite antigens in susceptible and resistant mice.

The specific antibody responses were compared among susceptible (A/Sn), moderately susceptible (Balb/c) and resistant (C57 BL/10J) mice infected with Trypanosoma cruzi (Y strain). Sera obtained during the second week of infection recognized a surface trypomastigote antigen of apparent Mr 80 kDa while displaying complex reactivity to surface epimastigote antigens. Complex trypomastigote antigens recognition was detected around the middle of the third week of infection. No major differences were observed along the infection, among the three strains of mice, neither in the patterns of surface antigen recognition by sera, nor in the titres of antibodies against blood trypomastigotes (lytic antibodies), tissue culture trypomastigotes or epimastigotes. On immunoblot analysis, however, IgG of the resistant strain displayed the most complex array of specificities against both trypo and epimastigote antigens, followed by the susceptible strain. IgM antibodies exhibited a more restricted antigen reactivity, in the three mouse strains studied. Balb/c sera (IgG and IgM) showed the least complex patterns of reactivity to antigens in the range of 30 kDa to 80 kDa. The onset of reactivity in the serum to trypomastigote surface antigens was also dependent on the parasite load to which the experimental animal was subjected.