Impact of gefitinib in early stage treatment on circulating cytokines and lymphocytes for patients with advanced non-small cell lung cancer

Figures & data

Figure 1 Gefitinib-induced changes of the peripheral lymphocyte subsets: (A) CD4⁺ T cells, (B) CD8⁺ T cells, (C) NK cells, and (D) CD4⁺CD25⁺ T cells.

Note: The changes of lymphocyte in terms of percent of PBMCs were analyzed for all patients (n=54) at baseline and 4 weeks after gefitinib treatment.
Abbreviation: PBMC, peripheral blood mononuclear cell.

Table 1 The characteristics of enrolled NSCLC patients (n=54)

Parameter case	N (%)
Gender
Female	28 (52)
Male	26 (48)
Smoking
Never	36 (67)
Ever	18 (33)
Stage
IIIB	3 (6)
IV	51 (94)
Pathology
ADC	48 (89)
Non-ADC	6 (11)
EGFR mutation status
Exon 19 del	33 (61)
Exon 21 L858R	17 (32)
Others*	4 (7)
Gefitinib efficacy
Partial response	41 (76)
Stable disease	9 (17)
Progression disease	4 (7)

Notes: Non-ADC includes four squamous carcinoma and two adenosquamous carcinoma.

* Other EGFR-activating mutations without EGFR exon T790M. Two cases harbor exon 18 (Gly719Ser and Gly719Ala) and another two cases with exon 21 (Leu861Gln), respectively.

Abbreviations: ADC, adenocarcinoma; EGFR, epidermal growth factor receptor; NSCLC, non-small cell lung cancer.

Figure 2 Gefitinib-induced decrease of circulating IL-6 and increase of INF-γ.

Notes: (A) IL-6, all patients. (B) IL-6, responders to gefitinib. (C) IL-6, nonresponders to gefitinib. (D) INF-γ, all patients. (E) INF-γ, responders to gefitinib. (F) nonresponders to gefitinib. All patients (n=54); responders to gefitinib (n=41); nonresponders to gefitinib (n=13).
Abbreviations: IL, interleukin; INF-γ, interferon-γ.

Table 2 The dynamic change of several immunological parameters and the efficacy of gefitinib

Parameters	Response (N)	Nonresponse (N)	χ^2	P-value
IL-2
Reduced	15	5	0.02	1.00
Nonreduced	26	8
IL-4
Reduced	13	6	0.90	0.51
Nonreduced	28	7
IL-6
Reduced	31	5	6.13	0.02
Nonreduced	10	8
IL-10
Elevated	18	4	–	0.52a
Nonelevated	23	9
INF-γ
Elevated	19	3	–	0.20a
Nonelevated	22	10
CD4^+CD25^+ T cells
Reduced	23	5	1.23	0.27
Nonreduced	18	8
Natural killer cells
Elevated	24	11	2.94	0.09
Nonelevated	17	2
CD4^+ T cells
Reduced	28	10	–	0.73a
Nonreduced	13	3
CD8^+ T cells
Elevated	22	7	<0.001	1.00
Nonelevated	19	6

Notes: The dynamic feature was classified according to the change trend that was observed in the study.

a P-value was calculated with Fisher’s exact test; all others were calaculated with Pearson chi-squared test.

Abbreviations: IL, interleukin; INF-γ, interferon-γ.

Figure 3 The prognostic value of IL-6 for PFS.

Note: Higher level (≥5.5 pg/mL) versus lower level (<5.5 pg/mL) of IL-6 at baseline.
Abbreviations: CI, confidence interval; HR, hazard ratio; IL, interleukin; PFS, progression-free survival.

Table 3 Univariate and multivariate survival analyses of 54 NSCLC patients

Variables	Category	Median PFS (months)	PFS analyses
			Univariate analyses			Multivariate analyses
			HR	95% CI	P-value	HR	95% CI	P-value
Gender	Female/male	11.8 vs 7.9	0.25	0.09–0.69	0.008	0.24	0.07–0.83	0.025
Age	<57/≥57 years	11.4 vs 7.9	0.39	0.17–0.90	0.028	0.24	0.08–0.66	0.006
Smoking history	No/yes	10.4 vs 7.9	0.35	0.14–0.91	0.031	1.58	0.50–5.26	0.452
EGFR status	Exon 19 del/others	11.8 vs 8.4	0.31	0.12–0.79	0.014	0.34	0.13–0.91	0.032
IL-6 level*	Lower/higher	11.4 vs 8.4	0.31	0.11–0.87	0.026	0.19	0.05–0.66	0.009
Response to gefitinib	PR/not PR	10.4 vs 10.2	0.45	0.18–1.12	0.086	0.17	0.05–0.53	0.002

Note:

* Patients were dichotomized on the basis of the median value (5.5 pg/mL) of baseline plasma IL-6.

Abbreviations: CI, confidence interval; EGFR, epidermal growth factor receptor; HR, hazard ratio; IL, interleukin; NSCLC, non-small cell lung cancer; PFS, progression-free survival; PR, partial response.

Figure S1 The changes of several circulating cytokines induced by gefitinib treatment.

Note: These were analyzed for all patients (n=54) with a baseline and 4 weeks after gefitinib treatment.

Abbreviation: IL, interleukin.

Figure S2 The prognostic value of IL-6 for progression-free survival.

Note: Higher level (≥1.7 pg/mL) versus lower level (<1.7 pg/mL) of IL-6 (A) and IL-6 reduced versus nonreduced after gefitinib treatment (B).

Abbreviation: IL, interleukin.

Figure S3 The expression of PD-L1 by immunohistochemical staining with a membranous pattern.

Note: Baseline staining (A) and 4 weeks after gefitinib treatment (B).

Abbreviation: PD-L1, programmed cell death ligand-1.

Table S1 Univariate and multivariate survival analyses involving plasma IL-6 change types

Variables	Category	Progression-free survival analyses
		Univariate analysis			Multivariate analysis
		HR	95% CI	P-value	HR	95% CI	P-value
Gender	Female/male	0.25	0.09–0.69	0.008	0.30	0.09–1.02	0.055
Age	<57/≥57 years	0.39	0.17–0.90	0.028	0.31	0.12–0.83	0.019
Smoking history	No/yes	0.35	0.14–0.91	0.031	0.75	0.21–2.67	0.654
EGFR status	Exon 19 del/others	0.31	0.12–0.79	0.014	0.28	0.10–0.77	0.013
IL-6 change types*	Reduced/nonreduced	0.36	0.11–1.14	0.083	0.52	0.14–1.95	0.335

Note:

* Patients were dichotomized based on the change type during gefitinib treatment.

Abbreviations: CI, confidence interval; EGFR, epidermal growth factor receptor; HR, hazard ratio; IL, interleukin.

Table S2 Univariate and multivariate survival analyses involving posttreatment plasma IL-6

Variables	Category	Progression-free survival analyses
		Univariate analysis			Multivariate analysis
		HR	95% CI	P-value	HR	95% CI	P-value
Gender	Female/male	0.25	0.09–0.69	0.008	0.23	0.06–0.82	0.024
Age	<57/≥57 years	0.39	0.17–0.90	0.028	0.30	0.11–0.81	0.017
Smoking history	No/yes	0.35	0.14–0.91	0.031	1.69	0.45–6.45	0.439
EGFR status	Exon 19 del/others	0.31	0.12–0.79	0.014	0.28	0.10–0.76	0.013
IL-6 level*	Lower/higher	0.57	0.25–1.33	0.200	0.47	0.16–1.33	0.153

Note:

* Patients were dichotomized based on the median value (1.7 pg/mL) of posttreatment plasma IL-6.

Abbreviations: CI, confidence interval; EGFR, epidermal growth factor receptor; HR, hazard ratio; IL, interleukin.

Table S3 The levels of peripheral lymphocytes at baseline or after 4 weeks of gefitinib treatment

Lymphocyte subgroup	Percent at baseline (%)	Percent after 4 weeks (%)	P-value
CD4^+ T cells	36.5±9.1	34.4±9.2	0.003
CD8^+ T cells	22.3±7.3	22.6±7.4	0.48
CD4/CD8 ratio	1.8±0.7	1.7±0.7	0.08
NK cells	21.3±10.0	23.7±10.9	0.005
CD4^+CD25^+ T cells	13.7±5.3	14.8±7.0	0.18

Note: The results of percent of lymphocytes of PBMC are presented as mean ± standard deviation.

Abbreviations: NK, natural killer; PBMC, peripheral blood mononuclear cell.