Multiplex immunofluorescence to measure dynamic changes in tumor-infiltrating lymphocytes and PD-L1 in early-stage breast cancer

Sanchez, Katherine; Kim, Isaac; Chun, Brie; Pucilowska, Joanna; Redmond, William L.; Urba, Walter J.; Martel, Maritza; Wu, Yaping; Campbell, Mary; Sun, Zhaoyu; Grunkemeier, Gary; Chang, Shu Ching; Bernard, Brady; Page, David B.

doi:10.1186/s13058-020-01378-4

Table 3 IRX-related effects on various immune cell subpopulations within stromal and tumor tissue compartments

From: Multiplex immunofluorescence to measure dynamic changes in tumor-infiltrating lymphocytes and PD-L1 in early-stage breast cancer

	Summary statistics		Unadjusted¹		Adjusted²
	Pre-IRX, mean density (SD)* (count/pixel)	Post-IRX, mean density (SD)* (count/pixel)	Fold change of mean density (95% CI)	p value	Fold change of mean density (95% CI)	p value
Stroma
Any PD-L1
T cell (any type)	0.85 (0.63)	1.27 (0.78)	1.8 (1.14 2.84)	0.015	2.01 (1.32 3.08)	0.001
Helper T cell	0.56 (0.47)	0.79 (0.59)	1.84 (0.97 3.48)	0.060	2.05 (1.14 3.70)	0.016
Cytotoxic T cell	0.22 (0.19)	0.42 (0.30)	2.18 (1.47 3.23)	< 0.001	2.55 (1.80 3.61)	< 0.001
Regulatory T cell	0.07 (0.05)	0.07 (0.06)	0.92 (0.57 1.49)	0.729	0.91 (0.60 1.39)	0.659
Macrophage	0.24 (0.18)	0.19 (0.12)	0.86 (0.54 1.37)	0.498	0.86 (0.54 1.38)	0.534
PD-L1-positive†
T cell (any type)	0.40 (0.57)	0.7 (0.59)	3.04 (1.48 6.23)	0.005	3.54 (1.86 6.76)	< 0.001
Helper T cell	0.28 (0.44)	0.47 (0.46)	3.31 (1.45 7.57)	0.008	3.58 (1.76 7.29)	< 0.001
Cytotoxic T cell	0.08 (0.11)	0.19 (0.21)	2.44 (1.28 4.63)	0.011	2.98 (1.74 5.08)	< 0.001
Regulatory T cell	0.05 (0.05)	0.05 (0.05)	1.39 (0.71 2.73)	0.311	1.38 (0.78 2.46)	0.272
Macrophage	0.12 (0.13)	0.14 (0.11)	1.66 (0.75 3.68)	0.192	1.63 (0.84 3.13)	0.147
Tumor
Any PD-L1
T cell (any type)	0.16 (0.17)	0.15 (0.15)	1.04 (0.61 1.8)	0.865	1.07 (0.65 1.77)	0.794
Helper T cell	0.10 (0.11)	0.07 (0.07)	0.81 (0.52 1.24)	0.304	0.77 (0.49 1.22)	0.268
Cytotoxic T cell	0.06 (0.05)	0.08 (0.09)	1.26 (0.81 1.94)	0.282	1.36 (0.91 2.05)	0.134
Regulatory T cell	0.02 (0.01)	0.02 (0.01)	0.72 (0.41 1.28)	0.242	0.79 (0.49 1.29)	0.346
Macrophage	0.04 (0.05)	0.03 (0.02)	0.71 (0.37 1.37)	0.282	0.65 (0.38 1.09)	0.105
PD-L1-positive†
T cell (any type)	0.11 (0.17)	0.09 (0.09)	1.42 (0.71 2.87)	0.295	1.50 (0.86 2.63)	0.151
Helper T cell	0.08 (0.13)	0.05 (0.05)	0.98 (0.51 1.91)	0.955	1.07 (0.56 2.04)	0.831
Cytotoxic T cell	0.04 (0.04)	0.05 (0.05)	1.63 (0.80 3.31)	0.155	1.68 (1.06 2.64)	0.026
Regulatory T cell	0.03 (0.05)	0.01 (0.01)	1.00 (0.49 2.05)	0.999	0.64 (0.32 1.29)	0.211
Macrophage	0.03 (0.05)	0.02 (0.02)	0.84 (0.51 1.39)	0.463	0.87 (0.51 1.49)	0.621
Stroma and tumor
PD-L1-positive†
PD-L1+ IC³	0.41 (0.53)	0.70 (0.54)	2.75 (1.36 5.53)	0.008	3.14 (1.68 5.87)	< 0.001

¹Paired t-test based on log-transformed density
²Poisson Generalized Linear Mixed-Effects Model (GLMM), with a log-linear effect of prevalence, an offset of log (area) (to make the expected number of cells proportional to area), the fixed effect of time (pre vs post), and two random effects of patients and ROI, to account for the variation in patients and ROI
*× 10^− 3; †PD-L1 > 2.6
T cell (any type) = combination of CD3+, CD8+, and FOXP3+ cell types; helper T cell = CD3+; cytotoxic T cell = CD8+; regulatory T cell = FOXP3+; macrophage = CD163
PD-L1 programmed death ligand 1, CI confidence interval

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