Analysis of T-Cell Clonality Pattern in Tumor Samples of Breast Cancer Patients

Purpose and methods A large body of experimental evidence has confirmed that different tumors, including breast carcinomas, can stimulate specific T-cell-mediated immune responses. In this study we have analyzed patterns of T-cell clonality in tumor samples of 54 breast cancer patients classified as lymph node negative, N0 (n=16), or lymph node positive, N+ (n=38). The clonality of T-cells was analyzed by the PCR-PAGE method. Results Monoclonal/oligoclonal (M/O) T-cell populations were found in 15 breast cancer patients, nine N+ and six N0. In all analyzed groups (N+ + N0, N+, N0) the incidence of relapse was not significantly different between patients with M/O and patients with polyclonal T-cells. Comparison of disease-free interval (DFI) between patients divided according to the presence of TCR? monoclonality/oligoclonality showed a marginally significant difference only in the group of N+ patients within the first 24 months of follow-up. Patients with a M/O T-cell population had a shorter DFI than patients with a polyclonal T-cell population. This difference was not observed when the complete follow-up period was considered in the same group of patients. Furthermore, there was no significant difference in overall survival (OS) between patients with M/O and patients with polyclonal T-cells. Conclusion Our results imply that tumor infiltrating T-cells are usually polyclonal. The pattern of T-cell clonality does not correlate with the incidence of relapse and the duration of DFI and OS in the analyzed groups of breast cancer patients, excluding N+ patients with M/O T-cells who had a shorter DFI in the first 24 months of follow-up. This observation suggests that polyclonal T-cell populations may provide a broader spectrum of T-cell-mediated antitumor response.

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Randomized cooperative study of perioperative chemotherapy in breast cancer.

Journal of Clinical Oncology ◽

10.1200/jco.1995.13.11.2712 ◽

1995 ◽

Vol 13 (11) ◽

pp. 2712-2721 ◽

Cited By ~ 22

Author(s):

M R Sertoli ◽

P Bruzzi ◽

P Pronzato ◽

P Queirolo ◽

D Amoroso ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Patients ◽

Preliminary Evidence ◽

Breast Cancer Patients ◽

Free Survival ◽

Node Negative ◽

Significant Survival ◽

Significant Difference ◽

Estrogen Receptor Negative

PURPOSE The aim of this multicentric randomized trial was to determine whether reducing the interval between surgery and chemotherapy improves the outcome of breast cancer patients. PATIENTS AND METHODS Between June 1985 and July 1992, 600 breast cancer patients, clinical stages T1-3A,N0-2,M0 were randomly assigned to a perioperative cycle (PC) of cyclophosphamide 600 mg/m2, epidoxorubicin 60 mg/m2, and fluorouracil 600 mg/m2 (CEF). Node-negative (N-) patients did not receive any further treatment. Node positive (N+) patients received 11 cycles if previously given PC, or 12 cycles of CEF alternated with cyclophosphamide 600 mg/m2, methotrexate 40 mg/m2, and fluorouracil 600 mg/m2 (CMF). In addition, N+ patients received concomitant or sequential 5-year tamoxifen therapy. RESULTS At a median follow-up duration of 5.7 years, no significant difference in survival (88% v 84%, P = .3) between the two treatment arms was seen. However, a difference of borderline significance in relapse-free survival (RFS; 76% v 70%, P = .053) was evident. A significant survival advantage for the PC arm was detected only in the estrogen receptor-negative (ER-) patients (P = .003). RFS was significantly improved in N- patients, postmenopausal patients, and ER- patients. Multivariate analyses show that pathologic tumor size, nodal status, receptor status, and treatment (only in ER- patients) are significantly correlated with survival and RFS. PC toxicity did not influence wound healing. CONCLUSION This study provides preliminary evidence that PC positively affects relapse rate and survival in some subgroups, namely, ER- patients.

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Tumor immune microenvironment (TiME) changes by multiplex IF staining in a pilot study of neoadjuvant talazoparib for early-stage breast cancer patients with a BRCA mutation.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.585 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. 585-585

Author(s):

Evthokia Hobbs ◽

Fei Yang ◽

Tapsi Kumar ◽

Alejandro Contreras ◽

Edwin Roger Parra Cuentas ◽

...

Keyword(s):

Breast Cancer ◽

T Cells ◽

T Cell ◽

Cancer Patients ◽

Early Stage ◽

Post Treatment ◽

Early Stage Breast Cancer ◽

Parp Inhibition ◽

Cytotoxic T Cell ◽

Breast Cancer Patients

585 Background: We previously reported a median tumor volume loss of 88% (range 30-98%) in 13 patients with early stage BRCA1/2 mutant breast cancer treated on a neoadjuvant trial of the PARP inhibitor talazoparib. The effects of PARP inhibition on immune aspects of the TiME in early-stage breast cancer has not been well described. The goal of this study was to evaluate the TiME in pre and post-treatment core biopsies from enrolled patients. Methods: Eleven paired core biopsies were available for examination. Tumor infiltrating lymphocytes (TILs) were quantified by H&E stained slides by a central pathologist. Specimens were assessed by multiplex immunofluorescence (mIF) using a panel of 6 biomarkers (PD-1, PD-L1, CD3, CD8, CD68 and CK) with the Opal 7-color Kit in LEICA BOND auto stainer, Vectra automated quantitative pathology imaging system and inForm software (PerkinElmer). Results: In the analyzed core biopsies, there was an increase in TILs evaluated by H&E in post-treatment compared to baseline (mean 36 vs 11%). By mIF there was an increase in CD3+ T cell and CD3+CD8+ cytotoxic T cell density in post-treatment samples compared to baseline, summarized in table. PD-L1 expression in tumor cells was rare in the cohort. There was no difference in CD3+PD-1+ or CD3+CD8+ PD-1+ lymphocytes in pre and post-treatment specimens. There was also no differences in macrophages (CD68+). Evaluation of immune phenotype and imaging response will be presented in the final analysis. Conclusions: This is the first study phenotyping the immune response to neoadjuvant talazoparib in BRCA-mutant breast cancer patients. In this small cohort, intratumoral and stromal CD3+ T cells and CD3+CD8+ cytotoxic T cells increased after two months of talazoparib. Clinical trial information: NCT02282345. [Table: see text]

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Analysis of naïve and memory CD4 and CD8 T cell populations in breast cancer patients receiving a HER2/neu peptide (E75) and GM-CSF vaccine

Cancer Immunology Immunotherapy ◽

10.1007/s00262-006-0188-9 ◽

2006 ◽

Vol 56 (2) ◽

pp. 135-146 ◽

Cited By ~ 27

Author(s):

Matthew T. Hueman ◽

Alexander Stojadinovic ◽

Catherine E. Storrer ◽

Zia A. Dehqanzada ◽

Jennifer M. Gurney ◽

...

Keyword(s):

Breast Cancer ◽

T Cell ◽

Cancer Patients ◽

Cd8 T Cell ◽

Cell Populations ◽

Breast Cancer Patients ◽

Gm Csf

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Prognostic significance of plasma osteopontin level in breast cancer patients

Forum of Clinical Oncology ◽

10.1515/fco-2015-0005 ◽

2015 ◽

Vol 6 (1) ◽

pp. 27-32

Author(s):

Hanan R. Nassar ◽

Alfred E. Namour ◽

Hanan E. Shafik ◽

Amr S. El Sayed ◽

Samar M. Kamel ◽

...

Keyword(s):

Breast Cancer ◽

Bone Metastasis ◽

Cancer Patients ◽

Biochemical Marker ◽

Menopausal Status ◽

P Value ◽

Breast Cancer Patients ◽

Pathological Type ◽

Significant Difference

Abstract Many studies have demonstrated that osteopontin (OPN) contributes functionally to aggressive behaviour in many tumours including breast cancer. This study aims to investigate its role as a simple biochemical marker easily measured in plasma of breast cancer patients to give an early signal for metastases and to detect its relationship to clinicopathological findings and survival. We measured plasma OPN, CA15.3 and serum alkaline phosphatase (ALP) activity in 55 patients, 28 with early stage breast cancer and 27 with bone metastasis out of whom 20 had metastasis in other sites. The median age at diagnosis for non-metastatic cases was 60 years (range 35-85) and for metastatic cases was 45.5 years (range 32-59). In the non-metastatic group, 78.57% of the patients were histologically graded as grades I and II and 21.43% as grade III tumours. In the metastatic group, 81.48% of the patients had grades I and II and 18.52% had grade III tumours; 54% of patients in the non-metastatic group were at stage II and 46% were at stage III at presentation. All patients of group II had bone metastasis, 33% had liver metastases, 25.9% had lung metastasis and 14.8% had lymph node metastasis. Patients with non-metastatic disease had a median OPN level of 55 ng/ml (range 54-150 ng/l), while those in the metastatic group had a median of 148.0 ng/l (range 56.0-156.0 ng/l), a difference which was statistically significant (P = 0.001). There was no statistically significant difference in the median levels of CA15.3 and ALP between both groups. The median OPN level was significantly higher with serum ALP level above 90, progesterone receptor (PR) status, bone and visceral metastasis. Median OPN was not affected significantly by menopausal status (P-value 0.3), tumour grade (P-value 0.3), estrogen receptor (ER) status (P-value 0.7), pathological type (P-value 0.42) or serum CA15.3 level (P-value 0.6). At the end of 12-year follow-up, 83% of the patients survived (92.3% in the non-metastatic versus 74.1% in the metastatic group). The estimated median survival for the whole study population at 12 years was 13 years (95% CI 8.144-17.856). The estimated median survival was 13 years (95% CI 0) and 12 years (95% CI 4.893-19.11) in patients with median OPN levels of <142 and ≥142, respectively, a difference which was not statistically significant (P = 0.343). No statistically significant difference in overall survival OS was noticed in relation to menopausal status (P = 0.7), pathological type (P = 0.4) and hormone receptor status (P = 0.3). At 6-year follow-up, it was found that OS was affected by the presence of visceral metastasis, tumour grade, serum plasma level of ALP and the serum level of CA15.3 (P = 0.0006, 0.007, 0.001 and 0.03, respectively). However, the presence of bone metastasis did not affect OS (P = 0.6). Osteopontin level can be a simple biochemical marker easily measured in plasma of breast cancer patients to give early signals for metastases, but not a prognostic factor for survival.

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Mice developing mammary tumors evolve T cell sequences shared with human breast cancer patients

10.1101/371260 ◽

2018 ◽

Cited By ~ 1

Author(s):

Miri Gordin ◽

Hagit Philip ◽

Alona Zilberberg ◽

Moriah Gidoni ◽

Raanan Margalit ◽

...

Keyword(s):

Breast Cancer ◽

T Cells ◽

T Cell ◽

Cancer Patients ◽

Human Breast Cancer ◽

Tumor Development ◽

Mammary Tumors ◽

Cell Receptor ◽

Breast Cancer Patients ◽

Human Breast

AbstractCancer immunotherapy by checkpoint blockade proves that an effective immune response to a tumor can be induced clinically. However, little is known about the evolution of tumor-associated T-cell receptor (TCR) repertoires without intervention. Here we studied TCR repertoire evolution in mice spontaneously developing mammary tumors; we sequenced peripheral blood alpha and beta TCRs of CD4+CD62L+CD44− T cells monthly for 8 months in 10 FVB/NJ mice transgenic at the Erbb2 locus, all developing tumors; 5 FVB/NJ mice without the transgene were age-matched controls. Sequences were either private (restricted to one mouse) or public (shared among mice); public sequences were either exclusive to the tumor group or inclusive among different groups. We now report that 1), public AA sequences were each encoded by many different nucleotide sequences (NT) recombinations (convergent recombination; CR); 2) mice developing tumors evolved tumor-exclusive public sequences, derived initially from private or from inclusive public sequences; and 3) tumor-exclusive public sequences in mice were also present among published public TCR sequences from human breast cancer patients. These cross-species tumor-exclusive TCR sequences manifested high CR; but the AA sequences shared by mice and humans did not share NT sequences. Thus, tumor-exclusive TCR AA sequences across species are selected from different NT recombination events. The roles of tumor-exclusive TCR repertoires in advancing or inhibiting tumor development and the effects of tumor immunotherapy on these T cells remain to be seen.

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Bone Marrow Micrometastases in Breast Cancer Patients: A Long-Term Follow-up Study

Clinical medicine Oncology ◽

10.1177/117955490800200404 ◽

2008 ◽

Vol 2 ◽

pp. 117955490800200

Author(s):

Annamaria Molino ◽

Monica Giovannini ◽

Rocco Micciolo ◽

Alessandra Auriemma ◽

Elena Fiorio ◽

...

Keyword(s):

Breast Cancer ◽

Bone Marrow ◽

Overall Survival ◽

Cancer Patients ◽

Breast Cancer Patients ◽

Primary Surgery ◽

Significant Difference ◽

Long Term Follow Up ◽

Disease Free

In 125 early breast cancer patients who underwent multiple bone marrow aspirates, there was no significant difference in terms of disease-free and overall survival after a median follow-up of 163 months between the patients with or without micrometastasis at the time of primary surgery. However, when the time-dependent evolution of the bone marrow aspirates was taken into account, some evidence for a longer disease-free and overall survival was found for the patients with negative bone marrow

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Quantification of Immune Variables from Liquid Biopsy in Breast Cancer Patients Links Vδ2+ γδ T Cell Alterations with Lymph Node Invasion

Cancers ◽

10.3390/cancers13030441 ◽

2021 ◽

Vol 13 (3) ◽

pp. 441

Author(s):

Stéphane Fattori ◽

Laurent Gorvel ◽

Samuel Granjeaud ◽

Philippe Rochigneux ◽

Marie-Sarah Rouvière ◽

...

Keyword(s):

Breast Cancer ◽

T Cells ◽

Lymph Node ◽

T Cell ◽

Cancer Patients ◽

Peripheral Blood ◽

Γδ T Cells ◽

Γδ T Cell ◽

Breast Cancer Patients ◽

Cancer Pathogenesis

The rationale for therapeutic targeting of Vδ2+ γδ T cells in breast cancer is strongly supported by in vitro and murine preclinical investigations, characterizing them as potent breast tumor cell killers and source of Th1-related cytokines, backing cytotoxic αβ T cells. Nonetheless, insights regarding Vδ2+ γδ T cell phenotypic alterations in human breast cancers are still lacking. This paucity of information is partly due to the challenging scarcity of these cells in surgical specimens. αβ T cell phenotypic alterations occurring in the tumor bed are detectable in the periphery and correlate with adverse clinical outcomes. Thus, we sought to determine through an exploratory study whether Vδ2+ γδ T cells phenotypic changes can be detected within breast cancer patients’ peripheral blood, along with association with tumor progression. By using mass cytometry, we quantified 130 immune variables from untreated breast cancer patients’ peripheral blood. Supervised analyses and dimensionality reduction algorithms evidenced circulating Vδ2+ γδ T cell phenotypic alterations already established at diagnosis. Foremost, terminally differentiated Vδ2+ γδ T cells displaying phenotypes of exhausted senescent T cells associated with lymph node involvement. Thereby, our results support Vδ2+ γδ T cells implication in breast cancer pathogenesis and progression, besides shedding light on liquid biopsies to monitor surrogate markers of tumor-infiltrating Vδ2+ γδ T cell antitumor activity.

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Clinical Evaluation of CA 15.3 in the Post-operative Follow-up of Breast Cancer Patients

The International Journal of Biological Markers ◽

10.1177/172460089000500104 ◽

1990 ◽

Vol 5 (1) ◽

pp. 22-26

Author(s):

M. Scaramuzzi ◽

C. Amorotti ◽

M. De Palma ◽

A.M. Falchi ◽

E. Baldini ◽

...

Keyword(s):

Breast Cancer ◽

Monoclonal Antibody ◽

Breast Carcinoma ◽

Cancer Patients ◽

Breast Cancer Patients ◽

Nodal Involvement ◽

Ca 15.3 ◽

Significant Difference ◽

Serum Test

In 265 patients operated for breast carcinoma the monoclonal antibody serum test CA 15.3 was predictive of metastatic diffusion of the disease. Its level increased in cases of distant metastasis with no significant difference between multiple and single sites (p = 0.014). The concentration of the marker was higher in 21 (23.8%) patients without nodal involvement and in 19 (27.5%) with nodal involvement (p = 0.193). Our study suggests that CA 15.3 may be an aid in the follow-up of patients with metastatic diffusion of breast cancer.

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Conventional versus hypofractionated postmastectomy radiotherapy: a report on long-term outcomes and late toxicity

Radiation Oncology ◽

10.1186/s13014-019-1378-x ◽

2019 ◽

Vol 14 (1) ◽

Cited By ~ 3

Author(s):

Imjai Chitapanarux ◽

Pitchayaponne Klunklin ◽

Attapol Pinitpatcharalert ◽

Patumrat Sripan ◽

Ekkasit Tharavichitkul ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Patients ◽

Late Toxicity ◽

Breast Cancer Patients ◽

Long Term Outcomes ◽

Postmastectomy Radiotherapy ◽

Free Survival ◽

Significant Difference

Abstract Objective We evaluated the long-term outcomes and late toxicity of conventional fractionated (CF) and hypofractionated (HF) postmastectomy radiotherapy (PMRT) in terms of locoregional recurrence-free survival (LRRFS), disease-free survival (DFS), overall survival (OS), and late toxicity. Methods A cohort of 1640 of breast cancer patients receiving PMRT between January 2004 and December 2014 were enrolled. Nine hundred eighty patients were treated with HF-PMRT: 2.65 Gy/fraction to a total of 42.4–53 Gy and 660 patients were treated with CF-PMRT: 2 Gy/fraction to a total of 50–60 Gy. Results The median follow-up time was 71.8 months (range 41.5–115.9 months). No significant difference was found in the rates of 5-year LRRFS, DFS, and OS of HF-PMRT vs CF-PMRT; 96% vs. 94% (p = 0.373), 70% vs. 72% (p = 0.849), and 73% vs. 74% (p = 0.463), respectively. We identified a cohort of 937 eligible breast cancer patients who could receive late toxicities assessment. With a median follow-up time of this patient cohort of 106.3 months (range 76–134 months), there was a significant higher incidence of grade 2 or more late skin (4% vs 1%) and subcutaneous (7% vs 2%) toxicity in patients treated with HF-PMRT vs CF-PMRT. Patients who received additional radiation boost were significantly higher in the HF-PMRT group. Grade 2 or more late RTOG/EORTC lung toxicity was significant lesser in HF-PMRT vs CF-PMRT (9% vs 16%). Grade 1 brachial plexopathy was also significant lesser in HF-PMRT vs CF-PMRT (2% vs 8%). Heart toxicity and lymphedema were similar in both groups. Conclusions HF-PMRT is feasible to deliver with comparable long-term efficacy to CF-PMRT. HF-PMRT had higher grade 2 or more skin and subcutaneous toxicity but less lung and brachial plexus toxicity.

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STAT3 Silencing and TLR7/8 Pathway Activation Repolarize and Suppress Myeloid-Derived Suppressor Cells From Breast Cancer Patients

Frontiers in Immunology ◽

10.3389/fimmu.2020.613215 ◽

2021 ◽

Vol 11 ◽

Author(s):

Elham Safarzadeh ◽

Ali Mohammadi ◽

Behzad Mansoori ◽

Pascal H. G. Duijf ◽

Shahryar Hashemzadeh ◽

...

Keyword(s):

Breast Cancer ◽

T Cells ◽

T Cell ◽

Cancer Cells ◽

Cancer Patients ◽

Suppressor Cells ◽

Myeloid Derived Suppressor Cells ◽

Breast Cancer Patients ◽

Hla Dr ◽

Stat3 Inhibition

Cancer cells escape immune destruction. From this perspective, myeloid-derived suppressor cells (MDSCs), which are immunosuppressive in various cancers including breast cancer (BC), are significant. However, the precise mechanisms are unknown. We isolated HLA-DR-CD33+ MDSCs and CD3+ T cells from BC patients’ peripheral blood and healthy donors through MACS and immunophenotyped by flow cytometry. Transfection of short-interfering RNAs and treatment with a TLR7/8 agonist altered pathway activities in vitro. Gene expression was analyzed using qRT-PCR, western blotting, and immunohistochemistry. Our findings showed an association between the progression of BC and increased levels of circulating HLA-DR-CD33+ MDSCs. These cells strongly suppress both autologous and analogous CD3+ T cell proliferation and enter the tumor microenvironment. We also identified increased STAT3 signaling and increased IDO and IL-10 expression in BC-derived MDSCs as immunosuppression mechanisms. Further, STAT3 inhibition and TLR7/8 pathway stimulation reduce the immunosuppressive activity of patient-derived MDSCs on T cells by inducing MDSC repolarization and differentiation into mature myeloid cells. This also alters the expression of critical cytokines and transcription factors in CD3+ T cells and, importantly, reduces breast cancer cells’ proliferation. Finally, while chemotherapy is able to significantly reduce circulating MDSCs’ level in patients with breast cancer, these MDSCs remained highly T cell-suppressive. We identified a novel molecular mechanism of MDSC-mediated immunosuppression. STAT3 inhibition and TLR7/8 pathway stimulation in MDSCs repolarize and suppress MDSCs from breast cancer patients. This offers new opportunities for BC immunotherapy.

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