Increased Stromal Infiltrating Lymphocytes are Associated with Circulating Tumor Cells and Metastatic Relapse in Breast Cancer Patients After Neoadjuvant Chemotherapy

Figures & data

Figure 1 Design of the study and time points of blood sampling and tissue sampling. ^#Surgery tissue samples were collected from patients and matched to healthy donors from whom biopsy tissue samples were sufficient for histologic evaluation.

Abbreviations: n, number; NAC, neoadjuvant chemotherapy; CTC, circulating tumor cell.

Table 1 Patient Characteristics and Correlation with CTC Detection Prior to and After Neoadjuvant Chemotherapy

	n (%)	Pre-NAC		Post-NAC
		CTC Positive (n=25)	P (χ^2)	CTC Positive (n=8)	P (χ^2)
Age
≦45	16 (53%)	13		3
>45	14 (47%)	12	1.000	5	0.417
Tumor size
T1 and T2	13 (43%)	12		5
T3 and T4	17 (57%)	13	0.355	3	0.102
Node
Negative	12 (40%)	10		3
Positive	18 (60%)	15	1.000	5	1.000
Tumor grade
1 and 2	12	11		2
3	18	14	0.622	6	0.419
Hormone receptors
ER or PR positive	16 (53%)	13		2
ER and PR negative	14 (47%)	12	1.000	6	0.101
HER2
Positive	14 (47%)	9		4
Negative	16 (53%)	16	0.014	4	1.000
Ki-67
≦20%	13 (43%)	11		2
>20%	17 (57%)	14	1.000	6	0.209

Notes: The bold value, p<0.05.

Abbreviations: n, number; NAC, neoadjuvant chemotherapy; CTC, circulating tumor cell; ER, estrogen receptor; PR, progesterone receptor.

Figure 2 (A) DFS of patients with or without pCR after NAC. (B) DFS of patients with or without detectable CTCs before the initiation of NAC. (C) DFS of patients with or without detectable CTCs after NAC.

Abbreviations: DFS, disease-free survival; NAC, neoadjuvant chemotherapy; CTC, circulating tumor cell.

Table 2 Association Between CTCs Detection and Recurrence in Breast Cancer Patients with Neoadjuvant Chemotherapy

	n (%)	Metastatic Relapse (n=13)	No Relapse (n=17)	P (χ^2)
Pre-NAC
Total	30
CTC positive	25	10	15
CTC negative	5	3	2	0.628
HR positive	15
CTC positive	12	2	10
CTC negative	3	1	2	0.516
HR negative	15
CTC positive	13	8	5
CTC negative	2	2	0	0.524
Post-NAC
Total	30
CTC positive	8	6	2
CTC negative	22	7	15	0.049
HR positive	15
CTC positive	2	1	1
CTC negative	13	2	11	0.371
HR negative	15
CTC positive	6	5	1
CTC negative	9	5	4	0.580

Notes: The bold value, p<0.05.

Abbreviations: n, number; NAC, neoadjuvant chemotherapy; CTC, circulating tumor cell; HR, hormone receptor.

Figure 3 Histologic patterns of lymphocyte infiltrate in a breast cancer case included in this study (400×). (A, C) iCD4⁺ and sCD4⁺ in core biopsies prior to NAC; (B, D) iCD8⁺ and sCD8⁺ in core biopsies prior to NAC; (E, G) iCD4⁺ and sCD4⁺ in surgical tissues after NAC; and (F, H) iCD8⁺ and sCD8⁺ in surgical tissues after NAC.

Abbreviations: iCD4⁺, intratumoral CD4⁺ tumor-infiltrating lymphocyte; iCD8⁺, intratumoral CD8⁺ tumor-infiltrating lymphocyte; sCD4⁺, stromal CD4⁺ tumor-infiltrating lymphocyte; sCD8⁺, stromal CD8⁺ tumor-infiltrating lymphocyte.

Table 3 Correlation of pCR or CTCs and Infiltrating Lymphocytes After Neoadjuvant Chemotherapy

	N1^#	Itil (N%)	P	Icd4^+ (N%)	P	Icd8^+ (N%)	P	N2	Stil (N%)	P	Scd4^+ (N%)	P	Scd8^+ (N%)	P
pCR
Yes	/	/		/		/		5	10 (10, 15)		6 (5, 10)		5 (3, 5)
No	/	/	/	/	/	/	/	14	12.5 (5, 30)	0.778	4.5 (2, 15)	0.402	3.5 (1, 10)	0.570
CTC detection
Positive	4	104 (13, 222.5)		48.5 (6.5, 96.5)		62.65 (6, 150.9)		6	25 (20, 50)		17.5 (10, 30)		9 (5, 15)
Negative	10	10.5 (3, 30)	0.320	3 (1, 5.6)	0.102	4.6 (1, 7.1)	0.224	13	10 (5, 15)	0.023	4 (2, 6)	0.010	2 (1, 5)	0.040
Metastatic relapse
Yes	6	41 (3, 185)		10.75 (2, 86)		19.6 (1, 114.3)		7	20 (10, 60)		15 (5, 40)		8 (3, 20)
No	8	10.6 (6.5, 22.25)	0.436	3 (1, 5.5)	0.218	4.6 (1, 7.05)	0.296	12	7.5 (5, 15)	0.028	3.5 (2, 8)	0.011	2.5 (1, 5)	0.110

Notes: ^#Among the 19 patients received TIL evaluation, iTILs after NAC cannot be detected for 5 patients who achieved pCR. The bold value, p<0.05.

Abbreviations: N, number; pCR, pathologic complete response; CTC, circulating tumor cell; iTIL, intratumoral tumor-infiltrating lymphocyte; sTIL, stromal tumor-infiltrating lymphocyte; iCD4⁺, intratumoral CD4⁺ tumor-infiltrating lymphocyte; iCD8⁺, intratumoral CD8⁺ tumor-infiltrating lymphocyte; sCD4⁺, stromal CD4⁺ tumor-infiltrating lymphocyte; sCD8⁺, stromal CD8⁺ tumor-infiltrating lymphocyte.