Monoclonal antibodies against mycobacterium avium/intracellulare

Ten hybridoma cell lines prodacing monoclonaI antibodies (Mabs) against M. avium/intracellulare (Mai) serotype 8 were raised by the fusion of BALB/c mouse myeloma cells (SPZ) to spleen cells from immunized BALB/c mice. The specificity of the monoclonal antibodies was defined using their differing abilities to bind to sonicates from a range of mycobacterial species and strains. Tbe Mabs showed strain and species specificity. Three Mabs bound only to Mai serotype 8 and 1 Mab bound only to Mai serotypes 8 and 16, the only serotypes tested. The results indicate that Mabs specific for Mai species and serotypes can be produced. These could be useful for serodiagnostic and for epidemiological purposes

Mycobacterium tuberculosis from chronic murine infections that grows in liquid but not on solid medium

BACKGROUND: Old, stationary cultures of Mycobacterium tuberculosis contain a majority of bacteria that can grow in broth cultures but cannot grow on solid medium plates. These may be in a non-replicating, dormant growth phase. We hypothesised that a similar population might be present in chronic, murine tuberculosis. METHODS: Estimates of the numbers of viable M. tuberculosis, strain H37Rv, in the spleens and lungs of mice in a 7-day acute infection and in a 10-month chronic infection were made by conventional plate counts and, as broth counts, by noting presence or absence of growth in serial replicate dilutions in liquid medium. RESULTS: Plate and broth counts in 6 mice gave similar mean values in the acute infection, 7 days after infection. However, the broth counts were much higher in 36 mice with a chronic infection at 10 months. Broth counts averaged 5.290 log(10 )cfu /organ from spleens and 5.523 log(10 )cfu/organ from lungs, while plate counts were 3.858 log(10 )cfu/organ from spleens and 3.662 log(10 )cfu/organ from lungs, indicating that the total bacterial population contained only 3.7% bacilli in spleens and 1.4% bacilli in lungs, capable of growth on plates. CONCLUSION: The proportion growing on plates might be a measure of the "dormancy" of the bacilli equally applicable to cultural and animal models

Comparative investigation of the pathogenicity of three Mycobacterium tuberculosis mutants defective in the synthesis of p-hydroxybenzoic acid derivatives.

p-Hydroxybenzoic acid derivatives (p-HBADs) are glycoconjugates secreted by all Mycobacterium tuberculosis isolates whose contribution to pathogenicity remains to be determined. The pathogenicity of three transposon mutants of M. tuberculosis deficient in the biosynthesis of some or all forms of p-HBADs was studied. Whilst the mutants grew similarly to the wild-type strain in macrophages and C57BL/6 mice, two of the mutants induced a more severe and diffuse inflammation in the lungs. The lack of production of some or all forms of p-HBADs in these two mutants also correlated with an increased secretion of the pro-inflammatory cytokines tumour-necrosis factor α, interleukin 6 and interleukin 12 in vivo. We propose that the loss of production of p-HBADs by tubercle bacilli results in their diminished ability to suppress the pro-inflammatory response to infection and that this ultimately provokes extensive pulmonary lesions in the C57BL/6 model of tuberculosis infection