Reinvigorating T-cell Response After BCG Vaccination

We investigated the potential of rIL-2, anti-KLRG-1 and anti-CTLA-4 antibody treatment to reinvigorate T-cell response in in vitro cultures at the 78 week time point after BCG vaccination of mice. We incubated lung or spleen cells with 10 ng mL⁻¹ of rIL-2, with or without WCL stimulation, in ELISPOT and ICS assays. In vitro KLRG-1 or CTLA-4 blockade using corresponding antibodies was also attempted [86] (link), [87] (link). Different concentrations of KLRG-1 antibody (2F1; eBiosceinces) up to 20 µg mL⁻¹ had no detectable effect on the cytokine response in these assays; however, 4F10 CTLA-4 antibody (BD-Biosciences) or purified Fab (20 µg mL⁻¹, using Pierce Fab Micro Preparation kit, Thermo Fisher) completely inhibited WCL-specific T-cell cytokine response suggesting inhibitory rather than blocking effect of this antibody [88] (link).

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Nandakumar S., Kannanganat S., Posey J.E., Amara R.R, & Sable S.B. (2014). Attrition of T-Cell Functions and Simultaneous Upregulation of Inhibitory Markers Correspond with the Waning of BCG-Induced Protection against Tuberculosis in Mice. PLoS ONE, 9(11), e113951.

Publication 2014

Pierce Fab Micro Preparation kit

Anti antibody Antibodies Antibody Assays Blocking antibody Cell cultures Cells Ctla 4 Cytokine Elispot Inhibitory Lung Mice Spleen T cell Vaccination bcg

Corresponding Organization :

Other organizations : Centers for Disease Control and Prevention, Emory University

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Variable analysis

independent variables

RIL-2 (10 ng mL^-1)
Anti-KLRG-1 antibody (2F1; up to 20 µg mL^-1)
Anti-CTLA-4 antibody (4F10; 20 µg mL^-1)
Anti-CTLA-4 Fab (20 µg mL^-1)

dependent variables

T-cell cytokine response (measured by ELISPOT and ICS assays)

control variables

Lung or spleen cells from BCG-vaccinated mice
Whole cell lysate (WCL) stimulation

controls

Positive control: Lung or spleen cells incubated with 10 ng mL^-1 of rIL-2
Negative control: Lung or spleen cells incubated without any treatment

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