{"doi":"10.21203/rs.3.rs-45672/v1","title":"CD4 T-Cell Immune Stimulation of HER2+ Breast Cancer Cells in Response to Trastuzumab In Vitro","abstract":"Abstract Introduction: The HER2+ tumor immune microenvironment is composed of macrophages, natural killer cells, and tumor infiltrating lymphocytes, which produce pro-inflammatory cytokines. Determining the effect of T-cells on HER2+ cancer cells during therapy could guide immunogenic therapies that trigger antibody-dependent cellular cytotoxicity. This study utilized longitudinal in vitro time-resolved microscopy imaging to measure T-cell influence on trastuzumab in HER2+ breast cancer. Methods: Fluorescently-labeled breast cancer cells (BT474, SKBR3, MDA-MB-453, and MDA-MB-231) were co-cultured with CD4+ T-cells (Jurkat cell line) and longitudinally imaged to quantify cancer cell viability when treated with trastuzumab (10, 25, 50 and 100 g/mL). The presence and timing of T-cell co-culturing was manipulated to determine immune stimulation of trastuzumab-treated HER2+ breast cancer. HER2 and TNF- expression were evaluated with western blot and ELISA, respectively. Significance was calculated using a two-tailed parametric t-test. Results: The viability of HER2+ cancer cells significantly decreased when exposed to 25 g/mL trastuzumab and T-cells, compared to cancer cells exposed to trastuzumab without T-cells (p = 0.01). The presence of T-cells significantly increased TNF- expression in trastuzumab-treated cancer cells (p = 0.02). Conversely, cancer cells treated with TNF- and trastuzumab had a similar decrease in viability as trastuzumab-treated cancer cells co-cultured with T-cells (p = 0.49). Conclusions: The presence of T-cells significantly increases the efficacy of targeted therapies and suggests trastuzumab may trigger immune mediated cytotoxicity. TNF- expression suggests cytokines may interact with trastuzumab-induced HER2 receptor blockade. Examining molecular mechanisms of breast cancer immune infiltration has the potential to improve response to targeted therapies.","journal":"Research Square","year":2020,"id":133629,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9599,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":578837,"name":"Amer Mansur","orcid":null,"position":1,"is_corresponding":false},{"id":430669,"name":"Kari J. Dugger","orcid":"0000-0002-9465-6128","position":2,"is_corresponding":false},{"id":578370,"name":"Tessa R. Davis","orcid":"0000-0003-0660-7850","position":3,"is_corresponding":false},{"id":430671,"name":"Grant Howard","orcid":"0000-0003-4731-7251","position":4,"is_corresponding":false},{"id":336776,"name":"Thomas E. Yankeelov","orcid":"0000-0001-6201-3913","position":5,"is_corresponding":false},{"id":430672,"name":"Anna G. Sorace","orcid":"0000-0002-3640-3966","position":6,"is_corresponding":false},{"id":430667,"name":"Patrick N. Song","orcid":"0000-0002-1545-0886","position":0,"is_corresponding":true}],"reference_count":30,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-18T23:16:17.577688Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}