- Volume 44, Issue 4, 1996

Production of a meningococcal vaccine capable of generating long-lasting immunity in all age groups is still a high priority worldwide. Iron-regulated outer-membrane proteins have attracted considerable attention in recent years and it has become increasingly evident that the meningococcal transferrin-binding proteins, TBP1 and TBP2, have characteristics compatible with a safe and broadly cross-reactive vaccine candidate. Both TBPs are surface-exposed and immunogenic in man and animals, and antibodies to their native structure are bactericidal to homologous and many heterologous strains. These include strains from various serogroups, serotypes and serosubtypes, with no obvious correlation between bactericidal activity and the identity of the strains or the molecular mass of the heterogeneous TBP2 molecule. A meningococcal vaccine based on, or enriched with, undenatured TBPs from one or more strains, in combination with conventional polysaccharide-based vaccines, might increase the spectrum of strains against which protection can be achieved to include serogroup B strains. In this review, the structure-function and immunological properties of TBP1 and TBP2 are discussed.

Fourteen commercial media supplied as pre-poured plates were compared with an ‘in-house’ selective medium for their ability to support the growth of 105 gonococcal isolates (representing a wide variety of serovars encountered in natural infection), 25 meningococcal and 20 Neisseria lactamica isolates, and to inhibit the growth of 71 isolates of non-pathogenic neisseriae and miscellaneous organisms. Only two of the prepoured plate media and the in-house selective medium yielded growth of duplicate cultures of all 105 gonococcal isolates after incubation for 24 h: one other medium provided growth of all the isolates after incubation for 48 h. The ability of the various media to suppress the growth of the 71 isolates of non-pathogenic neisseriae and miscellaneous organisms ranged from 97.2 to 71.8% of isolates inhibited. Of the four media that enabled growth of all the gonococcal strains, inhibition was 94.4% for the in-house medium, 85.9% and 80.3% for the two media on which all gonococci grew after 24 h and 71.8% for the medium on which all of the gonococci grew after 48 h. Failure of growth of gonococci was associated with: serogroup IA isolates (p < 0.001), AHU auxotype (p < 0.001) and the presence of vancomycin rather than lincomycin in the selective medium (p < 0.02). The use of 10% blood and a highly nutritious medium based on the original New York City (NYC) or modified New York City (MNYC) formulation were also important in supporting growth of gonococci. One of the main problems in lack of selectivity was a failure to inhibit the growth of yeasts. As effective inhibition of yeasts was obtained with other media containing the same concentration of amphotericin, failure may be due to batch variation of supplement, media preparation, or reduced shelf life of the media. None of the commercially available pre-poured media performed as well as the in-house medium despite the fact that some of the media were prepared to a very similar formula.

Helicobacter pylori is associated with various gastrointestinal disorders. Lethal photosensitisation was investigated as a possible technique for killing H. pylori which might offer a better alternative to antibiotics. The susceptibility of H. pylori to lethal photosensitisation was determined by mixing suspensions of H. pylori with various photosensitisers and plating out on blood agar before irradiation with low-power laser light. Five sensitisers were studied further by mixing them with H. pylori in a tissue-culture plate and counting survivors after irradiation as a function of laser exposure time, dye concentration and pre-irradiation time. Crystal violet and thionin were ineffective as sensitisers, but zones of inhibition appeared with methylene blue (MB), protoporphyrin IX (PPIX), haematoporphyrin derivative (HPD), toluidine blue O (TBO) and disulphonated aluminium phthalocyanine (S2). Laser light or sensitiser alone did not affect bacterial viability. S2 (100 μg/ml) with a laser light energy density of 16 J/cm2, HPD (100 μ/ml) with 160 J/cm2, MB (100 μg/ml) with 21 J/cm2, PPIX (150 μg/ml) with 320 J/cm2 and TBO (50 μg/ml) with 160 J/cm2 all reduced bacterial viability by > 99%. The killing of sensitised H. pylori by laser light offers a new approach to the treatment of localised infections when all colonised areas are accessible to light.

A significant increase in the incidence of isolates of methicillin-resistant Staphylococcus aureus (MRSA), that were also resistant to lincosamides and streptogramin A (LS A -MRSA), was observed in a French university hospital. Twenty-seven isolates from the outbreak were characterised, including 17 isolates from a plastic surgery ward and six control strains of MRSA. The strains were examined by antibiotyping and biotyping, and by three molecular methods: plasmid analysis, ribotyping and insertion sequence (IS) typing with IS256 sequence as a probe. Antibiotyping (five antibiotypes) was discriminatory because of the uncommon resistance phenotype of the epidemic strain. Biotyping (three biotypes), DNA plasmid analysis (four profiles) and ribotyping (two profiles) were poorly sensitive, in contrast to IS-typing (12 profiles). By the latter method, a coefficient of similarity (percentage similarity) compared to the predominant IS profile was calculated. Strains with a coefficient of similarity ⩾ 82% were considered as highly related to the epidemic strain, while those with a coefficient of similarity ⩽ 40% were regarded as distant. Results obtained with the five markers confirmed that an outbreak of hospital infection had occurred in the plastic surgery ward, with spread of the epidemic strain throughout the hospital.

There is a causal relationship between obesity-associated diabetes and an increased risk of infection. The ability of obese (fa/fa) Zucker rats, a model for non-insulin-dependent diabetes mellitus (NIDDM), to clear Candida albicans from the circulation and tissues was compared to that of lean (Fa/fa, Fa/Fa) Zucker rat controls as a measure of immune function. The ID50 necessary to establish tissue colonisation in lean Zucker rats was 1.18 log10 times greater than that determined for the obese Zucker rats. Nine days after intravenous (i.v.) injection of a yeast suspension, the organs of obese rats had a 10-fold greater yeast/g organ burden than did lean rats. The kidney was determined to be the primary target organ for colonisation. Germ-tube formation by C. albicans occurred at a rate 1.5 times faster in serum from obese rats than in serum from lean rats. Peritoneal polymorphonuclear leucocytes, resident macrophages and thioglycollateelicited macrophages from lean Zucker rats displayed a significantly higher ability to kill ingested yeast cells than analogous cell populations from obese Zucker rats.

In-vitro proteinase production by oral Candida albicans isolates from patients with and without HIV infection (18 isolates from each group) was assessed by image analysis of a plate assay, with bovine serum albumin (BSA) as a substrate. The effect of sub-minimal inhibitory concentrations (sub-MICs) of nystatin, amphotericin B, clotrimazole and miconazole on in-vitro proteinase production by these yeast isolates was also investigated. Proteinase production by C. albicans isolates from patients with HIV infection was significantly greater than production by those from individuals without infection. All 18 isolates from HIV-infected individuals produced proteinase, in comparison to 56% of isolates from uninfected individuals. Pre-exposure of C. albicans isolates (seven proteinase producers from each group) to 1/4 and 1/16 MICs of nystatin, amphotericin B, clotrimazole and miconazole resulted in decreased proteinase production in all isolates tested. However, after exposure to the four antimycotic agents, proteinase production was decreased to a significantly greater extent in isolates from uninfected individuals than in those with HIV disease. Furthermore, when the relative concentration effect of antimycotic agents on proteinase production was compared, C. albicans isolates from the HIV-free group demonstrated a salient dose-response relationship compared with the HIV-infected group. These results indicate that C. albicans from patients with HIV infection are significantly more proteolytic than those from individuals without the infection, and that polyenes and imidazoles curtail the proteolytic activity of all C. albicans isolates, albeit to a lesser extent in those from HIV-infected patients. It appears that HIV disease favours oral colonisation by more proteolytic C. albicans isolates, with resilient proteolytic activity.