Phenotypic profiling of HIV-responsive NK cells with mass cytometry

The purpose of this experiment is to profile the phenotype of the NK cells that are responsive to autologous HIV-infected CD4+ T cells. In this experiment, NK cells were co-cultured with autologous HIV-infected and mock-infected CD4+ T cells. The expression of NK cell surface receptors and functional markers were profiled with CyTOF analysis. By comparing the expression of NK cell surface receptors in the NK cells that are functional (expressing any of the functional markers CD107a, IFNgamma and TNFalpha) and not functional of responding to HIV-infected cells, we identified the NK cell receptors that are more predictive of the functional NK cells and evaluated if they are specific to HIV by comparing to the predictors of the functional NK cells when co-cultured with mock-infected CD4+ T cells.
Conclusion:
We identified multiple NK cell receptors to be predictive of functional responding NK cells and multiple NK cells receptors to be predictive of non-functional NK cells when NK cells were co-cultured with autologous HIV-infected CD4+ T cells. The top three predictors of functional responding NK cells are TIGIT, NKp30 and KIR3DL1, and the top three predictors of non-functional NK cells are CD16, NKp46, and CD62L. When the same analysis was performed on NK cells co-cultured with mock-infected CD4+ T cells, we found that many of the predictors of functional responding NK cells are similar, suggesting that the responding NK cell phenotype identified was not unique to HIV infection.
Notes:
The fcs files uploaded here are raw fcs files that have been normalized with EQ Beads. The fcs files were analyzed to gate on CD107a+, IFNgamma+ and TNFalpha+ cells within the NK cell population. Functional NK cells were identified with Boolean OR gate to include the cells that are stained positive for any of CD107a, IFNgamma, and TNFalpha. Non-functional NK cells were identified to be negative for all of CD107a, IFNgamma, and TNFalpha. Phenotypic analysis was performed on functional and non-functional NK cells.