FIG 3.

M. tuberculosis polar glycolipids retain the greatest specific activity for induction of γ₉δ₂ T cell inhibitory activity. M. tuberculosis H37Rv was cultured, and cells were harvested, irradiated, lyophilized, and enriched in bioactive product by the organic extraction. This extract was loaded onto a silica gel column and products were eluted with an increasing methanol gradient. (A) All the eluted fractions were tested for the ability to expand γ₉δ₂ T cells inhibitory for intracellular mycobacterial growth. (B) TLC analysis of biologically active extracts of M. tuberculosis. Shown are TLC results for M. tuberculosis H37Rv total lipid (100 μg) extracted with CHCl₃-CH₃OH-H₂O (10:10:3) and the 100% MeOH fraction (40 μg) eluted from the silica gel column loaded with dry extract. High-performance TLC sheets were developed in CHCl₃-CH₃OH-H₂O (56:38:10), and stained with α-naphthol (carbohydrate), ninhydrin (amino acids), or Dittmer-Lester reagent (phosphate).