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. Author manuscript; available in PMC: 2017 Jun 28.
Published in final edited form as: J Immunol. 2009 Mar 15;182(6):3668–3677. doi: 10.4049/jimmunol.0803579

FIGURE 5.

FIGURE 5

Depletion of ESAT-6 abrogates the inhibition of IFN-γ production by T cells. A, Preparations of recombinant histidine-tagged ESAT-6 were sham-depleted (lane 1) or depleted of ESAT-6 by incubation with a nickel resin (lane 2), and the efficiency of depletion was determined by Western blotting for ESAT-6 with mAb HYB76-8, after resolution by 15% SDS-PAGE and electroblotting to a nitrocellulose membrane. A representative result of two experiments is shown. B, Purified CD3+ cells from three healthy tuberculin-negative individuals were cultured for 1 h in medium alone, 3.3 μM of the sham-depleted recombinant ESAT-6, an equal volume of the ESAT-6-depleted preparation, or the buffer used for the depletion process, before stimulation with anti-CD3 and anti-CD28. After 48 h, IFN-γ levels in the culture supernatants were measured by ELISA. Mean values and SD are shown. *, p < 0.05, compared with the cells treated with the ESAT-6-depleted protein preparation.