Analysis of tumour-infiltrating lymphocytes reveals two new biologically different subgroups of breast ductal carcinoma in situ

Marie Beguinot, Marie-Melanie Dauplat, Fabrice Kwiatkowski, Guillaume Lebouedec, Lucie Tixier, Christophe Pomel, Frederique Penault-Llorca, Nina Radosevic-Robin, Marie Beguinot, Marie-Melanie Dauplat, Fabrice Kwiatkowski, Guillaume Lebouedec, Lucie Tixier, Christophe Pomel, Frederique Penault-Llorca, Nina Radosevic-Robin

Abstract

Background: Tumour-infiltrating lymphocytes (TILs) have been demonstrated to significantly influence prognosis and response to therapy of invasive breast cancer (IBC). Thus, it has been suggested that TIL density or/and immunophenotype could serve as biomarkers for selection of IBC patients for immunotherapy. However, much less is known about significance of TILs in breast ductal carcinoma in situ (DCIS).

Methods: We retrospectively investigated TIL density and immunophenotype in 96 pure DCIS and 35 microinvasive carcinomas (miCa). TIL density was assessed on H&E-stained breast biopsy sections as the percentage of tumour stromal area occupied by TILs, and classified into 4 grades: 0 (0%-9%), 1 (10-29%), 2 (30-49%) and 3 (50%-100%). TIL immunophenotype was assessed by immunohistochemistry for CD8, CD4, FoxP3, CD38 or CD20.

Results: Compared to pure DCIS, miCa contained significantly more cases with TIL density grade 3 (p = 0.028). Concordantly, CD8+, CD4+ and CD38+ cells were more numerous in miCa than in pure DCIS. In the pure DCIS subgroup with TIL density grades 2 and 3, all TIL subpopulations were more numerous than in the pure DCIS with TIL density grades 0 and 1, however the ratio between T-lymphocytes (CD8+ and CD4+) and B-lymphocytes (CD20+) was significantly lower (p = 0.029). On the other side, this ratio was significantly higher in miCa, in comparison with pure DCIS having TIL density grades 2 and 3 (p = 0.017). By cluster analysis of tumour cell pathobiological features we demonstrated similarity between miCa and the pure DCIS with TIL density grades 2 and 3. The only significant difference between those two categories was in the ratio of T- to B-TILs, higher in miCa.

Conclusion: Results indicate that TIL density level can distinguish 2 biologically different DCIS subgroups, one of which (DCIS with ≥30% TILs, the TIL-rich DCIS) is like miCa. Similarity of TIL-rich pure DCIS and miCa as well as the role of B-lymphocytes in DCIS invasiveness are worth further investigating with regards to the potential development of immunotherapy-based prevention of DCIS progression.

Keywords: breast; cancer; ductal; in situ; lymphocytes; microinvasive.

Conflict of interest statement

Ethics approval and consent to participate

All patients whose tissue and clinical data were used for this study have signed, already at admission to the Jean Perrin Cancer Centre, that they agree with the use of their tissues and clinical data for research purposes. The study presented in this article was approved by Comité d’Ethique des Centres d’Investigation Clinique de l’inter-région Rhône-Alpes-Auvergne. Approval reference: 00005921/CE-CIC-GREN-17-14.

Consent for publication

Not applicable.

Competing interests

The authors declare having no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Examples of TIL density grades. a) a case representing TIL density grade 0 (TILs are estimated to occupy 2% of the stromal area). H&E, original magnification 4×, scale bar 250 μm; b) a case representing TIL density grade 1 (TILs are estimated to occupy 20% of the stromal area). H&E, original magnification 5×, scale bar 200 μm; c) a case representing TIL density grade 2 (TILs are estimated to occupy 40% of the stromal area). H&E, original magnification 5×, scale bar 200 μm; d) a case representing TIL density grade 3 (TILs are estimated to occupy 60% of the stromal area). H&E, original magnification 5×, scale bar 200 μm. The circles show the areas sampled for tissue microarray construction
Fig. 2
Fig. 2
Cluster analysis of distribution of TIL-based subcategories according to the tumour cell pathobiological characteristics. Red colour represents higher values/presence of a parameter; green colour represents lower values/absence of a parameter. For molecular subtypes: green = luminal, brown = HER2+, red = triple negative. The numbers in blue, green and orange are IDs of the cases analysed

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