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PubMed1990

Distribution of a protein antigenically related to the major anaerobically induced gonococcal outer membrane protein among other Neisseria species.

The Pan 1 protein of Neisseria gonorrhoeae is a novel 54-kDa outer membrane protein expressed only when gonococci are grown in the absence of oxygen. It is a major antigen recognized by sera from patients with gonococcal infection. We raised mouse monospecific polyclonal antiserum to gel-purified Pan 1 from gonococcal strain F62. The antiserum was broadly cross-reactive among gonococcal strains; all strains tested reacted in immunoblot analysis proportionate to the amount of Pan 1 visible in silver-stained sodium dodecyl sulfate (SDS)-polyacrylamide gels. In immunoblot experiments, N. lactamica and N. cinerea reacted very strongly to the anti-Pan 1 antiserum, whereas N. sicca, N. flava, and N. mucosa did not react at all. The other commensals tested, N. subflava and N. perflava, exhibited only a minor reaction. These results correlated with the apparent amount of Pan 1 seen on SDS-polyacrylamide gels of outer membranes. SDS-polyacrylamide gel analysis of six meningococcal strains revealed no visible anaerobically induced outer membrane proteins, and the subsequent immunoblots showed only slight or no reaction to the anti-Pan 1 antibody. In the four meningococcal strains that did react slightly with the antiserum, a Pan 1-like protein was seen only in anaerobically grown cells. Thus, meningococci did not express Pan 1 at levels comparable to that found in gonococci; however, when Pan 1 was expressed in meningococcal strains, it was oxygen regulated. This is the first example of a protein found in the gonococcal outer membrane that, under identical growth conditions, is not expressed at similar levels in the meningococcus.

Anaerobiosis↗

PubMed1982

[Salmonella meningitis].

Cultures were done for 14.838 cerebro spinal fluid specimens (CSF) during 1977-1980. Positive results were obtained from 950 specimens. These were identified as 352 Klebsiella-Enterobacter (37.0%), 133 Pseudomonas (14.%), 96 Salmonella (10.1%), 65 Pneumococcus (6,9%), 51 Escherichia coli 5,4%), 50 Meningococcus (.3%), Haemophilus influenzae type B (1.0%) and 127 various miscellaneous bacteria. During the same period of time, total of Salmonella isolations from various sources were 865. These specimens were 488 stool (57.0%), 160 blood (18.6% 7) 96 CSF (11.2%) so forth. The majority of these Salmonella strains were of typhimurium serotype carrying a known antibiotic resistence plasmid. This plasmid, belonging to the fime compatibility group, present in the Salmonella typhimurium strains isolated in Middle-East countries like Iran, Israel and others, conferred resistance to the carrying strain against antibiotics like ampicillin, kanamycin, chloramphenicol, streptomycin, sulphamide and tetracycline. As started to show up in Ankara too, addition of trimethoprim and gentamicin resistences to this markers were reported. These bacteria were also causing epidemics in pediatric wards of some South America countries which develops with hing rate of bacteremia and meningitis complications as it is in our hospital.

Blood↗

PubMed1999

Analysis of the human Ig isotype response to lactoferrin binding protein A from Neisseria meningitidis.

An effective vaccine for serogroup B meningococci has yet to be developed and attention has turned to subcapsular antigens of the meningococcus as possible vaccine candidates. Iron binding proteins are being studied, with most interest focused on the transferrin binding proteins (TbpA and TbpB) and the ferric binding protein (FbpA). This study describes the purification of lactoferrin binding protein A (LbpA) from two meningococcal strains and assesses the human isotype-specific serum antibody response to these proteins in patients with proven meningococcal disease due to a range of phenotypes. Overall, fewer than 50% of sera contained IgG that recognised LbpA isolated from either strain and this antibody response was not uniform between the two proteins. There was some evidence that the antibody response varied between meningococcal phenotypes. This study demonstrates that LbpA does not induce a highly cross-reactive antibody response, indicating that it is unlikely to be an effective vaccine antigen.

Antibodies, Bacterial↗

PubMed2005

Serum serotype-specific pneumococcal anticapsular immunoglobulin g concentrations after immunization with a 9-valent conjugate pneumococcal vaccine correlate with nasopharyngeal acquisition of pneumococcus.

BACKGROUND: Immunization with pneumococcal conjugate vaccines (PCVs) reduces nasopharyngeal colonization by Streptococcus pneumoniae. We attempted to correlate postvaccination serum serotype-specific pneumococcal anticapsular immunoglobulin (Ig) G concentrations with new acquisitions of vaccine-type (VT) serotypes and the VT-related serotype 6A. METHODS: A total of 132 day care center attendees aged 12-35 months received a 9-valent PCV (PnCRM9) and were followed for 2 years for new nasopharyngeal acquisitions of S. pneumoniae. A total of 132 control subjects received a meningococcus type C conjugate vaccine. Serum serotype-specific pneumococcal anticapsular IgG concentrations were determined at 1 month after complete immunization. RESULTS: A logistic regression model of the probability of having a new acquisition of S. pneumoniae (for serotypes 9V, 14, 19F, and 23F) as a function of the IgG concentration showed a negative coefficient, indicating that higher IgG concentrations led to a decreasing probability of having a new acquisition, and achieved statistical significance for serotypes 14 and 19F. Similarly, a new acquisition of serotype 6A was shown to be significantly inversely related to the anti-6B IgG concentration. An effect of the IgG concentration on duration of carriage was not demonstrated. CONCLUSION: The magnitude of herd protection against S. pneumoniae provided by a PCV may depend on the magnitude of IgG concentrations.

Antibodies, Bacterial↗

PubMedDate not supplied

Meningococcus: a menace in Cork?

A review of laboratory isolates and notifications of meningococcal disease in the Cork area was conducted for the period 1989-93. The study aimed to describe the epidemiology of meningococcal disease in the area. The incidence of meningococcal disease is high in the Cork area and has been increasing since 1991 with a peak incidence of laboratory confirmed cases of 6.5 per 100,000 in 1993. In the five year review period 113 notifications of meningococcal disease were identified of which 61 (54%) were laboratory confirmed and 52 (46%) were clinically diagnosed only. All laboratory confirmed cases had been notified to the local Director of Community Care/Medical Officer of Health. Group C organisms comprised two-thirds of isolates in 1992 and 1993. Comparison of regional and national incidence rates must be based on laboratory confirmed cases as the criteria for diagnosis and completeness of notifications may vary. A National Infectious Disease Surveillance Centre is vital for monitoring trends and for the coordinated development of a national policy on control and prevention of meningococcal disease.

Age Distribution↗

PubMed1996

Meningococcal disease in Los Angeles County, California, and among men in the county jails.

BACKGROUND: From January through March 1993, there were 54 cases of meningococcal disease in Los Angeles County, California, of which 9 occurred among men incarcerated in the county's jail system, which was 40 percent above capacity at the time. Several of the 45 patients from the community had had contact with men recently released from a county jail. METHODS: We interviewed patients from the community (n=42) and neighborhood controls matched with the patients for age, race, and ethnic group (n=84) about potential exposures. We collected and cultured pharyngeal swabs for Neisseria meningitidis from men entering the central jail (n=162), men leaving the central jail (n=379), members of the jail staff (n=121), and patients at a community health center (n=214). Meningococcal isolates were identified by serotyping and multilocus enzyme electrophoresis. RESULTS: The presence of community-acquired meningococcal disease was strongly associated with exposure to a person who had been in or worked at one of the county jails (multivariate matched odds ratio, 18.5; 95 percent confidence interval, 3.8 to 90.8; P<0.001). Pharyngeal carriage of meningococcus was significantly more frequent among men released from jail (19 percent) or entering jail (17 percent) than among workers at the jails (3 percent) or community residents seen at the clinic (1 percent). Among men entering jail, those who had previously been incarcerated were more often carriers than those who had not (21 percent vs. 7 percent, P=0.03). Of the isolates from nine community residents with serogroup C meningococcal disease, eight were the same strain as that isolated from the eight inmates with serogroup C disease. CONCLUSIONS: In this outbreak of meningococcal disease in Los Angeles County, nearly half of community residents with the disease had contact with persons who had been in a county jail. The high rates of carriage among recidivists and released inmates suggests that the men became meningococcal carriers while in jail.

Carrier State↗

PubMed2005

Interlaboratory comparison of PCR-based identification and genogrouping of Neisseria meningitidis.

Twenty clinical samples (18 cerebrospinal fluid samples and 2 articular fluid samples) were sent to 11 meningococcus reference centers located in 11 different countries. Ten of these laboratories are participating in the EU-MenNet program (a European Union-funded program) and are members of the European Monitoring Group on Meningococci. The remaining laboratory was located in Burkina Faso. Neisseria meningitidis was sought by detecting several meningococcus-specific genes (crgA, ctrA, 16S rRNA, and porA). The PCR-based nonculture method for the detection of N. meningitidis gave similar results between participants with a mean sensitivity and specificity of 89.7 and 92.7%, respectively. Most of the laboratories also performed genogrouping assays (siaD and mynB/sacC). The performance of genogrouping was more variable between laboratories, with a mean sensitivity of 72.7%. Genogroup B gave the best correlation between participants, as all laboratories routinely perform this PCR. The results for genogroups A and W135 were less similar between the eight participating laboratories that performed these PCRs.

Adolescent↗

PubMed2002

Many carried meningococci lack the genes required for capsule synthesis and transport.

Of 830 Neisseria meningitidis isolates obtained from healthy carriers in Bavaria, Germany, 136 (16.4%) lacked the operons necessary for the synthesis, lipid modification, and transport of capsular polysaccharide. These operons were replaced by a non-coding intergenic region either 113 or 114 bp in length, termed here the capsule null locus (cnl). Comparisons of the nucleotide sequence of this region in the meningococcus and its acapsulate relatives, Neisseria gonorrhoeae and Neisseria lactamica, revealed six distinct sequence variants (cnl-1 to cnl-6), with a total of 10 nucleotide substitutions and three indels. With the exception of one 4 bp insertion, which was unique to a gonococcal isolate, all of the individual sequence changes were present in the N. lactamica isolates examined. The meningococcal isolates with a cnl belonged to one of four otherwise genetically diverse genetic groupings: the ST-53 and ST-1117 complexes (75 isolates); the ST-845 complex (12 isolates); the ST-198 and 1136 complexes (46 isolates), and the ST-44 complex (one isolate). These data demonstrated that a substantial proportion of carried meningococci were incapable of capsule production, that the cnl circulated within Neisseria populations by horizontal genetic exchange, and that the expression of a polysaccharide capsule was not a requirement for person-to-person transmission of certain meningococcal lineages.

Bacterial Capsules↗

PubMed2005

Serotype incidence and antibiotic susceptibility of Streptococcus pneumoniae causing invasive disease in Scotland, 1999-2002.

Pneumococcal disease remains an important cause of invasive and non-invasive disease in Scotland and elsewhere. The Scottish Meningococcus and Pneumococcus Reference Laboratory receives isolates of Streptococcus pneumoniae from diagnostic laboratories around Scotland. Here, the serogroups/types and antibiotic-susceptibility patterns of invasive isolates received between 1999 and 2002 are described. There were a total of 1741 invasive isolates received, the most common serogroups/types being 14 (19.8 %), 9 (10.2 %), 6 (8.3 %), 19 (7.9 %), 23 (7.9 %), 4 (6.5 %), 8 (6.4 %), 3 (5.7 %), 1 (3.8 %), 7 (3.8 %) and 18 (3.4 %). Importantly, serotypes 7 and 8 are not represented in the 7-, 9- and 11-valent pneumococcal conjugate polysaccharide vaccines. There were 67 (3.8 %) isolates considered penicillin non-susceptible, although no penicillin resistance (MIC > or = 0.002 mg ml(-1)) was recorded. One hundred and ninety-four (11.1 %) isolates, predominantly of serotype 14, were resistant to erythromycin, and 12 (0.7 %) were resistant to ciprofloxacin. This information provides an important dataset that will prove essential prior to and during the implementation of pneumococcal conjugate vaccines in the UK.

Anti-Bacterial Agents↗

PubMed2005

Neisseria meningitidis lactate permease is required for nasopharyngeal colonization.

Neisseria meningitidis is a human specific pathogen that is part of the normal nasopharyngeal flora. Little is known about the metabolic constraints on survival of the meningococcus during colonization of the upper airways. Here we show that glucose and lactate, both carbon energy sources for meningococcal growth, are present in millimolar concentrations within nasopharyngeal tissue. We used a mutant defective for the uptake of lactate (C311DeltalctP) to investigate the contribution of this energy source during colonization. Explants of nasopharyngeal tissue were inoculated with the wild-type strain (C311) and C311DeltalctP; the mutant was recovered at significantly lower levels (P = 0.01) than C311 18 h later. This defect was not due to changes in the expression of adhesins or initial adhesion in C311DeltalctP to epithelial cells. Instead, lactate appears to be important energy source for the bacterium during colonization and is necessary for growth of the bacterium in nasopharyngeal tissue. Studies with other strains defective for the uptake of specific nutrients should provide valuable information about the environment in which N. meningitidis persists during carriage.

Adhesins, Bacterial↗

PubMed2001

Prospects offered by genome studies for combating meningococcal disease by vaccination.

Meningococcal disease was first recognised and Neisseria meningitidis isolated as the causative agent over 100 years ago, but despite more than a century of research, attempts to eliminate this distressing illness have so far been thwarted. The main problem lies in the fact that N. meningitidis usually exists as a harmless commensal inhabitant of the human nasopharynx, the pathogenic state being the exception rather than the norm. As man is its only host, the meningococcus is uniquely adapted to this ecological niche and has evolved an array of mechanisms for evading clearance by the human immune response. Progress has been made in combating the disease by developing vaccines that target specific pathogenic serogroups of meningococci. However, a fully comprehensive vaccine that protects against all pathogenic strains is still just beyond reach. The publication of the genome sequences of two meningococcal strains, one each from serogroups A and B and the imminent completion of a third illustrates the extent of the problems to be overcome, namely the vast array of genetic mechanisms for the generation of meningococcal diversity. Fortunately, genome studies also provide new hope for solutions to these problems in the potential for a greater understanding of meningococcal pathogenesis and possibilities for the identification of new vaccine candidates. This review describes some of the approaches that are currently being used to exploit the information from meningococcal genome sequences and seeks to identify future prospects for combating meningococcal disease.

Clinical Trials as Topic↗

PubMed1985

Carriage of Neisseria meningitidis and Neisseria lactamica in a school population during an epidemic period in Spain.

A study was made of the incidence of Neisseria meningitidis and N. lactamica in a school population; 2470 children aged between 5 and 7 years were studied from four schools in Alcala de Henares (Madrid). Nasopharyngeal swabs were taken in June, November and March, between 1979 and 1983. In all the surveys except one, the proportion of carriers of N. lactamica was higher than that of N. meningitidis, reaching a ratio of about 2:1 in the complete study. The predominant serogroup of meningococcus found was B (41%), with nongroupable strains reaching 43%. A study of serotypes within group B showed a predominance of nontypable strains (48.5%), while those strains considered to be most virulent (types 2 and 1, 8, 15) reached 40%. Eighteen per cent of N. lactamica strains were observed to agglutinate with antimeningococcal sera whilst the remainder of the strains were rough. When these strains were studied with the antiserum-agar technique, using antimeningococcal sera, a high percentage of strains cross-reacted with the meningococci. The susceptibility of strains to sulphadiazine, penicillin, ampicillin, chloramphenicol, rifampicin and spiramycin was determined. Finally an analysis was made of the effect that an elevated colonization rate of N. lactamica might have on colonization by meningococci. The necessity of using fine epidemiological markers in tracing virulent strains in a population at risk is stressed. Selective prophylactic measures are also necessary.

Anti-Bacterial Agents↗

PubMed2003

Pneumococcal bacteraemia: clinical and microbiological epidemiology in Dundee, Scotland.

OBJECTIVE: To report the incidence of drug resistant Streptococcus pneumoniae isolated from blood culture in Dundee, Scotland. We shall also review the clinical and laboratory findings in these cases. METHODS: A retrospective review was undertaken of all cases of S. pneumoniae bacteraemia identified in our local area during a three year period from August 1st, 1997 to July 31st, 2000 (107 cases.) Data was obtained from patient medical records, blood culture reports and results of Stoke's disk testing. Many organisms were also sent to the Scottish Meningococcus and Pneumococcus Reference Laboratory for serogrouping and determination of the minimum inhibitory concentration of common antibiotics. RESULTS: Annual incidence of bacteraemia was approximately 15.9-17.8 per 100000 population. Mortality was 33% (34 and 30% for those with pneumonia or meningitis, respectively). No relationship was seen between patient age and overall mortality. Factors relating to increased mortality were a high respiratory rate (p=0.01), high blood urea level (p=0.05) and the presence of confusion (p<0.01) on admission to hospital. The incidence of penicillin resistant S. pneumoniae was 7%, all of these isolates having low level resistance. Macrolide resistance was 8%. Neither were found to be increasing over the three year period. The most common serogroups were 23 (18%) and 14 (12%). CONCLUSIONS: The incidence of penicillin resistant S. pneumoniae isolated from blood culture in Dundee, Scotland, is similar to the UK average and did not appear to be rising between 1997 and 2000.

Adolescent↗

PubMed1983

[Evaluation of ceftriaxone (R013-9904) in the treatment of septicemia caused by gram-negative bacilli].

Thirty-four patients admitted for a suspicion of septicaemia were treated with ceftriaxone, a third generation cephalosporin. Ceftriaxone was administered IV at a dose of 2 grams daily, either as a single injection of 2 g or as two injections of 1 g each. 43 organisms were isolated from the blood of the 34 patients: 20 E.coli; 5 Klebsiella; 2 Salmonella; 2 indole positives Proteus; 1 indole negative Proteus; 1 Enterobacter; 2 Acinetobacter; 3 Staphylococcus aureus; 2 Streptococcus; 1 Enterococcus; 1 Meningococcus and 3 anaerobes. The MIC of the enterobacteria for ceftriaxone ranged from less than or equal to 0.097 microgram/ml to 1.56 microgram/ml. Only two staphylococci, one Acinetobacter and the enterococcus were resistant to the drug. Serum assays of ceftriaxone were performed on the second day of treatment for 15 patients. Within the group treated with a single dosis of 2 g per day, the blood levels were: maximal level (10 min, after the injection): 175 to 460 micrograms/ml (average: 315), minimal level (before injection) 12 to 100 micrograms/ml (average 53). Among the patients treated with two doses of 1 g per day, we obtained: maximal levels 121 to 260 micrograms/ml (average 178), minimal levels 31 to 70 micrograms/ml (average 52). A clinically favorable evolution was obtained for 29 patients (85%). In the cases of 4 patients, the antibiotherapy had to be adapted in view of the susceptibility of the organisms isolated. Ceftriaxone was very well tolerated. The only observed secondary effect was a drug fever occurring in 3 patients who remained febrile in spite of a general improvement.

Bacteria↗

PubMed2005

Solution structure of the immunodominant domain of protective antigen GNA1870 of Neisseria meningitidis.

GNA1870, a 28-kDa surface-exposed lipoprotein of Neisseria meningitidis recently discovered by reverse vaccinology, is one of the most potent antigens of Meningococcus and a promising candidate for a universal vaccine against a devastating disease. Previous studies of epitope mapping and genetic characterization identified residues critical for bactericidal response within the C-terminal domain of the molecule. To elucidate the conformation of protective epitopes, we used NMR spectroscopy to obtain the solution structure of the immunodominant 18-kDa C-terminal portion of GNA1870. The structure consists of an eight-stranded antiparallel beta-barrel overlaid by a short alpha-helix with an unstructured N-terminal end. Residues previously shown to be important for antibody recognition were mapped on loops facing the same ridge of the molecule. The sequence similarity of GNA1870 with members of the bacterial transferrin receptor family allows one to predict the folding of this class of well known bacterial antigens, providing the basis for the rational engineering of high affinity B cell epitopes.

Amino Acid Sequence↗

PubMed2004

Expression of heterologous antigens in commensal Neisseria spp.: preservation of conformational epitopes with vaccine potential.

Commensal neisseriae share with Neisseria meningitidis (meningococcus) a tendency towards overproduction of the bacterial outer envelope, leading to the formation and release during growth of outer membrane vesicles (OMVs). OMVs from both meningococci and commensal neisseriae have shown promise as vaccines to protect against meningococcal disease. We report here the successful expression at high levels of heterologous proteins in commensal neisseriae and the display, in its native conformation, of one meningococcal outer membrane protein vaccine candidate, NspA, in OMVs prepared from such a recombinant Neisseria flavescens strain. These NspA-containing OMVs conferred protection against otherwise lethal intraperitoneal challenge of mice with N. meningitidis serogroup B, and sera raised against them mediated opsonophagocytosis of meningococcal strains expressing this antigen. This development promises to facilitate the design of novel vaccines containing membrane protein antigens that are otherwise difficult to present in native conformation that provide cross-protective efficacy in the prevention of meningococcal disease.

Animals↗

PubMed2001

Epidemiological pattern of meningococcal disease in Valencia, Spain. Impact of a mass immunization campaign with meningococcal C polysaccharide vaccine.

The objective of this study was to define the epidemiological pattern of meningococcal disease in the autonomous region of Valencia, Spain, and the impact of a mass immunization campaign against serogroup C meningococcus. Data were obtained from a prospective surveillance program for invasive bacterial diseases in children < 15 y of age that began in the Valencia region on 1 December, 1995. During the period 1996-98, 213 cases of meningococcal disease were detected, representing an annual incidence of 11.3/100,000 children < 15 y. Serogroup C accounted for 31% and 38.5% of cases in 1996 and 1997, respectively (annual incidences of 2.9 and 5.4 cases/100,000 children < 15 y). An immunization campaign with the meningococcal C polysaccharide vaccine, which included all persons between 18 months and 19 y of age, began in late 1997 (vaccination coverage of 86%). In 1998, the annual incidence of meningococcal C disease fell to 1.4 cases per 100,000 children < 15 y of age. These results mirror the increase in the reported incidence of serogroup C meningococcal disease in Spain in the 1990s, a trend that was reversed after the introduction of the mass vaccination campaign. Meningococcal polysaccharide vaccine seems to be an effective public health tool for the management of this serious communicable disease.

Adolescent↗

PubMed2001

Semiautomation of multilocus sequence typing for the characterization of clinical isolates of Neisseria meningitidis.

The Scottish Meningococcus and Pneumococcus Reference Laboratory (SMPRL) provides a national service for the laboratory confirmation of meningococcal and pneumococcal disease in Scotland. Part of this service includes the serogrouping of meningococcal isolates followed by typing and subtyping. The procedures for this are labor-intensive but important for the identification of linked cases and the surveillance of disease so that effective public health measures can be taken. However, different strains of meningococci, such as those within the electrophoretic type 37 complex, occurring during case clusters of disease are now indistinguishable by current methods. The SMPRL has started using multilocus sequence typing (MLST) as a routine method for the characterization of isolates of Neisseria meningitidis. MLST produces nucleotide sequence data of seven housekeeping genes providing results that are useful for public health management. However, the method is laborious and time-consuming and therefore lends itself towards automation. The SMPRL therefore developed a semiautomated method for MLST using a 96-well format liquid handler and an automated DNA sequencer. Semiautomated MLST is now provided as a reference service for Scotland. This work describes the methodology required for the characterization of N. meningitidis and highlights its usefulness for public health intervention.

Automation↗