Figure legends
Figure 1. Flow diagram of eligible study population. A total of
68 patients were selected with advanced solid tumors (including 42 lung
cancers, 14 liver cancers, nine colon cancers and three renal
carcinomas) who had experienced disease progression after at least one
previous course of tumor-appropriate treatment for advanced or
metastatic disease.
Figure 2. Naïve T cells from patients were related to CIK cell
therapy response. (A) Cluster analysis of flow cytometry data acquired
from patients before CIK cell therapy. The measured values of subsets of
absolute blood cell numbers are displayed as heatmaps, which are colored
per subset according to mean-centered and sigma-normalized data
(Z-scores). The top-bars indicate the PD, SD or PR/CR of each
individual. (B and C) Flow cytometry analysis of sorted human
CD45RA+CCR7+CD62L+naïve CD8+ and CD4+ T cells acquired
from patinaiveents before CIK cels therapy. Numbers indicate the
percentage of cells in the gates. Graphs show PD vs SD vs PR/CR
expression in the
CD45RA+CCR7+CD62L+CD8+or CD4+ gate. (D, F, G, H) Percentages and absolute
numbers of CD8+ (D and F) and CD4+(G and H) T cells in subsets of patients with different clinical
prognoses before CIK cells therapy (obtained as described in panel C).
Figure 3. Light microscopic representation of CIK cells from
patients with high and low numbers of naïve T cells on days 3, 5, 9 and
13 after amplification in vitro . (B) Quantification of
proliferation of T cells after amplification in vitro . Cells were
placed in 96-well plates (4,000 cells/well); the first bar represents
the unstimulated control cells, the second bar represents stimulated T
cells. (C) The frequency of
PD-1+TIM3+CD8+cells in CIK cell expansions from patients with high and low numbers of
naïve T cells at the indicated time points. (D) The frequency of
Ki67+CD8+ cells in CIK cell
expansions from patients with high and low numbers of naïve T cells at
the indicated time points. (E) Cytolytic activity of non-activated CIK
cells expanded from patients with high and low numbers of naïve T cells
in response to their respective autologous tumor cells, which were
obtained from two NSCLC patients. E:T ratio, effector cell to target
cell ratio. (F, G, H) The frequency of
PD-1+TIM3+CD8+cells at each time point from PD (F), SD (G) or PR/CR (H) patients
before or after amplification in vitro . (I) The frequency of
IFN-γ+CD3+CD56+cells before and after amplification in vitro from patients in
different response categories.
Figure 4. The CIK cells derived from patients with different
frequencies of naïve T cells drive the response to therapy. (A, B, C &
D) The frequency of
IL-2+CD3+CD56+(A),
IFN-γ+CD3+CD56+(B),
PD-1+CD3+CD56+ (C)
and Ki67+CD3+CD56+cells (D) before and on days 5, 10, 15, 20 and 25 after CIK cell therapy
in different response categories.
Figure 5. Patients
with high numbers of naïve T cells show more potent and durable
antitumor responses in vivo. (A) Best percent change in target
lesion tumor burden from baseline. Data only includes patients with
baseline target lesions and one or more post-baseline target lesion
assessments with no missing values (n = 66). Maximum percent reductions
in target lesion tumor burden from baseline across all tumor assessments
before subsequent therapy were used. Positive change in tumor burden
indicates tumor growth; negative change in tumor burden indicates tumor
reduction. Horizontal lines denote 30% decrease and 20% increase. Not
all reductions of ≥30% from baseline are partial responses (i.e.,
decrease in target lesion tumor burden but new or progressive nontarget
lesions). (B) Percent change in target lesion tumor burden from baseline
over time. Data only includes patients with baseline target lesions and
one or more post-baseline target lesion assessments with no missing
values (n = 66). Horizontal lines denote 30% decrease, 20% increase
and no change. (C) MRI showing that a 43-year-old CRC patient with brain
metastases who had a high naïve T count experienced a CR of the brain
after eight cycles of CIK cell infusion (Top). Chest CT scans showing a
51-year-old NSCLC patient with high numbers of naïve T cells experienced
PR of the lung after four cycles of CIK cell infusion (middle). Chest CT
scans showing a 48-year-old NSCLC patient with a high naïve T cell count
experienced PR of the lung after four cycles of CIK cell infusion
(bottom); the arrows show regression of ascites. (D) Time to and
duration of response. CR, complete response; DOR, duration of response.
(E) Progression-free survival (PFS). Data for PFS are based on an August
2018 database lock. Symbols denote censored observations.