Figure 5
Analysis of absolute white blood cell count (WBC), neutrophil count and flow-cytometric assessment of lymphocytes, T cell subsets and natural killer cells in patient 1 during the treatment phase. After preparative chemotherapy (CTX) (cyclophosphamide 350 mg/m2 and fludarabine 20 mg/m2) the patient was reconstituted with 0.9 × 1010 autologous PBMC (5.05 × 107 CD3-positive cells/kg) followed by 5 cycles of the autologous tumor cell vaccine. All of the determined CD3-positive cell subsets (CD4, CD8, CD4CD25) were affected by cyclophosphomide and fludarabine. As seen in all patients, neutrophil counts recovered to pre-chemotherapy levels within 30 days. Recovery of different T cell subsets (CD4, CD8, CD4CD25) was slower than normalization of neutrophil counts in all patients but varied inter-individually. The post-chemotherapy increase in CD4 numbers followed the same kinetics as the other subsets with no extended depression as one might expect following fludarabine treatment. Patient 1 experienced an extended depression of CD19-positive B lymphocytes, whereas NK cells (CD3-CD16+CD56+) recovered within 30 days (as observed in all patients). Additional leukaphereses were harvested prior to preparative chemotherapy and after the vaccination phase.