Bacillus Calmette-Guerin vaccination of human newborns induces T cells with complex cytokine and phenotypic profiles

The immune response to vaccination with bacillus Calmette-Guerin (BCG), the only tuberculosis vaccine available, has not been fully characterized. We used multiparameter flow cytometry to examine specific T cell cytokine production and phenotypic profiles in blood from 10-wk-old infants routinely vaccinated with BCG at birth. Ex vivo stimulation of whole blood with BCG for 12 h induced expression of predominantly IFN-gamma, IL-2, and TNF-alpha in CD4(+) T cells in seven distinct cytokine combinations. IL-4 and IL-10 expression was detected in CD4(+) T cells at low frequencies and only in cells that did not coexpress type 1 cytokines. Specific CD8(+) T cells were less frequent than CD4(+) T cells and produced mainly IFN-gamma and/or IL-2 and less TNF-alpha, IL-4, and IL-10. Importantly, many mycobacteria-specific CD4(+) and CD8(+) T cells did not produce IFN-gamma. The predominant phenotype of BM-specific type I T cells was that of effector cells, i.e., CD45RA(-)CCR7(-)CD27(+), which may reflect persistence of Mycobacterium bovis BCG in infants until 10 wk of age. Among five phenotypic patterns of CD4(+) T cells, central memory cells were more likely to be IL-2(+) and effector cells were more likely to be IFN-gamma(+). We concluded that neonatal vaccination with BCG induces T cells with a complex pattern of cytokine expression and phenotypes. Measuring IFN-gamma production alone underestimates the magnitude and complexity of the host cytokine response to BCG vaccination and may not be an optimal readout in studies of BCG and novel tuberculosis vaccination.