Figure 1.

Evaluation of nuclear factor-κβ (NF-κB) essential modulator (NEMO) mutation cryptococcal meningoencephalitis (CM) Case 1 and anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) autoantibodies in CM Case 2 with idiopathic CD4⁺ lymphopenia. (A) Sanger deoxyribonucleic acid sequencing of polymerase chain reaction product demonstrates X-linked G>A 169 mutation in IKBKG gene, which encodes E57K NEMO in CM Case 1. (B) Trends in CD4⁺ T lymphocyte count over time for CM Case 1 with E57K NEMO mutation depict persistent CD4⁺ lymphopenia despite recovery of CD8⁺ T cells postsplenectomy. (C) Flow cytometry T-cell receptor (TCR) stimulation experiments using different titers of anti-CD3 (10 μg/mL, 5 μg/mL, 1 μg/mL), and unstimulated upon CD3⁺ negatively selected T lymphocytes with fixed costimulation with anti-CD28 (2 μg/mL) (eBioscience/Affymetrix, San Diego, CA) over 36 hours reveals significant increasing perecentage of interferon (IFN)γ induction among CD4⁺ T lymphocytes with increasing anti-CD3CD28 titer among controls but less amounts among CM Case 1 with E57K NEMO and CM and other patients with the same mutation but no CM. Numerical values in boxes represent %CD4⁺ expressing IFNγ. (D) Increased induction of phosphorylated NF-κB (RelA/p65) corresponds to more IkBα degradation among CD4⁺ T lymphocytes from control compared with CM Case 1 as predicted by NEMO mutation. The IkBα-phosphorylated state in stimulated case is at level of unstimulated control. The TCR stimulation was performed at fixed anti-CD3 (5 μg/mL) and anti-CD28 (2 μg/mL). Numerical values in boxes represent %CD4⁺ expressing NF-κB (RelA/p65) phosphorylated on serine at position 529. (E) The GM-CSF-induced STAT-5 phosphorylation in normal CD14⁺ monocytes in the presence of 10% normal or patient plasma from CM Case 2.