Table 1.
Application of multiplex staining in clinical immune profiling studies.
| Tumour type | Marker panel | Summary | Reference |
|---|---|---|---|
| Prognostic studies | |||
| Breast cancer (HER2+ and TNBC) | CD4, CD8, CD20, FOXP3, CD68, PanCK | Higher B cell infiltration was associated with better overall survival | [62] |
| Breast cancer | CD8, CD103, CD69, PanCK, DAPI | Higher tissue-resident memory T cell infiltration was associated with better recurrence-free survival | [63] |
| Breast cancer (TNBC) | CD4, CD8, FOXP3, CD20, CD33, PD-1 | Higher PD-1+ CD8+ T cells, PD-1+CD4+ T cells were associated with better prognosis | [64] |
| Breast cancer (TNBC) | CD4, CD8, FOXP3, PD-1, PD-L1 | CD4/PD-L1, CD8/PD-1, and CD8/PD-L1 double-positive TILs were significantly associated with recurrence | [65] |
| Pancreatic ductal adenocarcinomas | CD8, KRT7 | High density of CD8+ T cells in tumour centre was associated with improved survival | [66] |
| Endometrial cancer | CD8, CD4, FOXP3 | High Treg counts and Treg/CD8+ ratios were significantly associated with worse distant metastasis-free survival | [67] |
| Lung cancer (NSCLC) | Panel 1: CD4, CD38, CD68, FOXP3, CD20 Panel 2: CD8, PD-L1, CD163, CD68, CD133 | Patients were classified into three subtypes based on their unique immune composition (immune activated, immune defected and immune exempted), where immune-activated patients showed the longest disease-free survival | [68] |
| Lung cancer (NSCLC) | CD3, CD8, CD20 | Higher CD3+ and CD8+ infiltration was associated with better outcome | [69] |
| Gastric cancer | CD68, CD163, CD206, IRF8, PD-L1 | Higher CD163+ (CD206−) tumour-associated macrophage density with high CD68 expression was associated with better patient survival | [70] |
| Gastric cancer | CD4, CD8, FOXP3, PD-1, PD-L1, TIM3 | Higher levels of CD8, PD-1, and PD-L1 following NAC were associated with better overall survival | [71] |
| Predictive studies | |||
| Melanoma | CD8, FOXP3, SOX10 PD-1, PD-L1 | CD8+ cells within 20 μm of a melanoma cell were predictive of PD-1-based immunotherapy. | [72] |
| Melanoma | CD4, CD8, CD20, CD3, GZMB, Ki67 | Pretreatment lymphocytic infiltration (CD3, CD8) was indicative of anti-PD-1 response | [73] |
| Lung cancer (NSCLC) | CD3, CD8, CD4, PD-1, CD57, FOXP3, CD25, Granzyme B | CD8+ T cells lacking PD-1 inhibitory receptor positively impacted nivolumab-treated patient survival | [74] |
| Lung cancer (NSCLC) | Panel 1 - AE1/AE3, PD-L1, CD3, CD4, CD8, and CD68 Panel 2 - AE1/AE3, PD-1, granzyme B, FOXP3, CD45RO, CD57. |
Higher intratumoural T helper cell and macrophage levels were associated with chemotherapy outcome | [75] |
| Breast cancer | CD3, CD20, CD8 | CD3, CD8, and CD20 infiltration were predictive of NAC response | [76] |
| Breast cancer (HER2+) | CD8, CD4, CD20, CD68, FoxP3, CK | CD4+, CD8+, CD20+ s-TILs, CD20+ s-TILs were independently associated with higher pCR in patients treated with neoadjuvant anti-HER2 therapy | [77] |
| Breast cancer | PD-L1, PanCK | PD-L1 was predictive of NAC response | [78] |
| Gastric cancer | CD8, PD-1, PD-L1, TIM-3, LAG-3, CD4, CD20, FoxP3, CTLA-4, CD68, CD163, CD66b, HLA-DR, STING | Multi-marker protein signature predicts anti-PD-1/PD-L1 therapy response | [79] |
| Lung cancer (NSCLC) | PD-L1, CD68, CD8 | High level of PD-1 in macrophages was linked with higher CD8+ cell infiltration and better survival outcome in PD-1 pathway blockade therapy | [80] |
pCR, pathological complete response; s-TIL, stromal tumour-infiltrating lymphocyte; TNBC, triple-negative breast cancer.