TABLE 1.
Effect of vaccination and in vitro stimulation on MCP-1 mRNA in guinea plg alveolar macrophages
| Vaccination statusa | Treatment of cells in vitrob | Mean ± SEM of fold induction at indicated time poststimulationc
|
||
|---|---|---|---|---|
| 3 h | 12 h | 24 h | ||
| Nonvaccinated | Control | 1.0 ± 0 | 1.0 ± 0 | 1.0 ± 0 |
| LPS | 2.2 ± 0.2 | 2.1 ± 0.4 | 3.2 ± 1.3 | |
| H37Ra | 2.2 ± 0.8 | 2.9 ± 0.4 | 2.6 ± 1.3 | |
| H37Rv | 2.8 ± 0.8 | 2.4 ± 1.5 | 0.9 ± 0.7 | |
| BCG vaccinated | Control | 2.1 ± 1.3 | 2.7 ± 0.8 | 1.3 ± 1.0 |
| LPS | 4.5 ± 2.8 | 4.1 ± 2.2 | 2.8 ± 3.2 | |
| H37Ra | 4.2 ± 2.8 | 4.1 ± 2.1 | 9.9 ± 10.6 | |
| H37Rv | 2.6 ± 1.8 | 3.1 ± 1.5 | 3.3 ± 3.1 | |
Vaccinated animals received a subcutaneous injection of 0.1 ml of M. bovis BCG vaccine (Danish 1331: Statens Seruminstitut, Copenhagen, Denmark) in the left inguinal region as described in Materials and Methods.
Alveolar macrophages isolated from nonvaccinated and BCG-vaccinated guinea pigs were infected with two laboratory strains of M. tuberculosis, H37Ra and H37Rv (MOI, 1:1), or were cultured with lipopolysaccharide (LPS, 10 μg/ml.) from E. coli. Control alveolar macrophages were incubated in complete media alone.
In vitro cultured alveolar macrophages from nonvaccinated and BCG-vaccinated guinea pigs (four animals per treatment group) were infected with live M. tuberculosis H37Ra or H37Rv or cultured with or without LPS for 3, 12, and 24 h. Total RNA from the macrophages was collected, reverse transcribed to cDNA, and detected by real-time PCR. Differences between groups were compared by Student's one-tailed t test, assuming equal variances. P values of <0.05 were considered significant.