Prolonged overall survival in gastric cancer patients after adoptive immunotherapy

Table 1 Expanded activated autologous lymphocyte cell properties

Property	Value
Proliferation time (d)	13.55 ± 1.25
Number of cells before proliferation (106/L)	7.65 ± 1.52
Number of cells after proliferation (109/L)	8.76 ± 1.82
Proliferation multiplicity	1156.57 ± 167.88
Survival rate of cells	97.57% ± 0.94%

Table 2 Phenotypes of lymphocytes before and after in vitro culture

Phenotype	Before (%)	After (%)	P valuea
CD3+	67.39 ± 7.55	96.71 ± 3.85	< 0.001
CD3+CD4+	33.22 ± 11.57	10.17 ± 8.83	< 0.001
CD3+CD8+	27.85 ± 8.51	79.33 ± 13.58	< 0.001
CD8+CD28+	13.19 ± 6.08	51.01 ± 15.34	< 0.001
CD3-CD16+/CD56+	28.12 ± 12.10	5.56 ± 6.48	< 0.001
CD3+CD16+/CD56+	9.19 ± 6.42	12.73 ± 7.65	0.019
CD45RA+	60.84 ± 8.26	9.74 ± 5.87	< 0.001
CD45RO+	46.53 ± 10.01	94.32 ± 4.57	< 0.001
CD4+CD25+	5.61 ± 4.19	3.17 ± 2.15	0.002
CD4+CD25+CD127-	1.00 ± 0.46	0.94 ± 1.50	0.448
CD29+	62.15 ± 8.77	93.04 ± 3.98	< 0.001
CD4+CD29+	20.11 ± 5.97	12.36 ± 7.55	< 0.001

^aP < 0.05 vs the phenotype before in vitro culture.

Table 3 Basic clinical characteristics of patients treated with or without expanded activated autologous lymphocyte adoptive immunotherapy

Characteristic	Patients treated with EAALs (n)	Patients treated without EAALs (n)	P value
Patient number	42	42
Age (yr)
< 60	24	22	0.661
≥ 60	18	20
Sex
Male	34	33	0.786
Female	8	9
Stage
I and II	10	9	0.909
IIIa and IIIb	12	11
IIIc and IV	20	22
Surgery
Yes	33	37	0.242
No	9	5
Radiotherapy
Yes	8	4	0.212
No	34	38
Chemotherapy
≤ 6 cycles	19	24	0.275
> 6 cycles	23	18

EAALs: Expanded activated autologous lymphocytes.

Table 4 Comparison of survival rates in gastric cancer patients with or without expanded activated autologous lymphocyte immunotherapy

Group	n	1 yr survival rate (%)(95%CI)	2-yr survival rate (%)(95%CI)	3-yr survival rate (%)(95%CI)	4-yr survival rate (%)(95%CI)	5-yr survival rate (%)(95%CI)
Immunotherapy group	42	80.95	54.35	41.1	36.54	25.05
		(69.08-92.83)	(39.16-69.54)	(25.79-56.41)	(20.52-52.55)	(7.78-42.33)
Control group	42	61.9	33.33	16.67	11.11	11.11
		(47.22-76.59)	(19.08-47.59)	(5.40-27.94)	(0-22.75)	(0-22.75)
P value		> 0.05	> 0.05	> 0.05	> 0.05	> 0.05

Table 5 Subgroup analysis

Subgroup	Median OS (mo)		RR (95%CI)	P value
	Immunotherapy group	Control group
Age (yr)
< 60	25.8	13.4	0.65 (0.333-1.272)	0.209
≥ 60	46.5	13.9	0.502 (0.233-1.079)	0.078
Gender
Male	20.0	15.0	0.439 (0.146-1.32)	0.143
Female	30.5	13.9	0.625 (0.355-1.099)	0.103
Clinical stage
I and II	60.0	62.2	1.212 (0.293-5.013)	0.791
IIIa and IIIb	49.3	28.4	0.358 (0.128-1.003)	0.051
IIIc and IV	14.0	10.0	0.484 (0.251-0.933)	0.03a
Surgery
Yes	36.0	15.4	0.561 (0.319-0.986)	0.045a
No	19.7	9.4	0.265 (0.069-1.015)	0.053
Radiotherapy
Yes	60.0	43.5	0.382 (0.094-1.55)	0.178
No	22.0	13.4	0.624 (0.364-1.069)	0.086
Chemotherapy cycles
≤ 6 cycles	27.0	10.0	0.544 (0.255-1.159)	0.115
> 6 cycles	30.5	18.4	0.554 (0.28-1.093)	0.089

^aP < 0.05 was considered statistically significant.

Table 6 COX multivariate regression analysis of gastric cancer patients

Factor	Wald	P value	HR	95%CI
Gender	2.527	0.112	1.723	0.881-3.369
Age	1.799	0.180	0.659	0.359-1.212
Clinical stage	41.852	< 0.001
Clinical stage (1)	37.267	< 0.001a	0.050	0.019-0.131
Clinical stage (2)	22.155	< 0.001a	0.172	0.083-0.358
Surgery	0.474	0.491	0.777	0.378-1.595
Chemotherapy cycles	0.069	0.793	1.083	0.597-1.966
Radiotherapy	0.581	0.446	1.386	0.599-3.209
Application of EAALs	7.819	0.005a	2.249	1.274-3.969

^aP < 0.05 was considered statistically significant. EAALs: Expanded activated autologous lymphocytes.

Table 7 Adverse events n (%)

Event	Immunotherapy patients (n = 42)1	Number of EAAL cellular transfers (n = 242)
Fever	8 (19.05)	26 (10.74)
Chill	4 (9.52)	13 (5.37)
Headache	3 (7.14)	5 (2.07)
Nausea	2 (4.76)	2 (0.83)
Itching	1 (2.38)	1 (0.41)
Rash	1 (2.38)	1 (0.41)
Tachycardia	1 (2.38)	1 (0.41)
Diarrhea	1 (2.38)	1 (0.41)

¹Five patients showed more than one type of event attributed to immunotherapy. EAAL: Expanded activated autologous lymphocyte.