Expression of CD8 marker and proliferation of peripheral blood lymphocyte of multiparous women by Co-culture with breast tumor cell, MDA-231 and MCF-7 lines and comparison with nulliparous women

Document Type : Original Article


1 MSc Student of Immunology, Reproductive Endocrinology Research Center, Research Institute for Endocrine Sciences and Metabolism, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

2 Assistant Professor, Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

3 Professor, Department of Immunology, Reproductive Endocrinology Research Center, Research Institute for Endocrine Sciences and Metabolism, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

4 Professor, Research Institute for Endocrine Sciences and Metabolism, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.


Introduction: Low frequency of tumor incidence and metastatic case of breast cancer in multiparous mothers accompanied with this hypothesis that the incidence of microchimerism could be one of the reason of this phenomenon. Therefore, in this study, to investigate mononuclear cells response of peripheral blood adjacent in culture with tumor cell lines of breast cancer, it was attempted to implement an in-vitro co-culture model.
Methods: From 48 multiparous mothers and nulliparous women, Peripheral Blood Mononuclear Cells)PBMC( were purified and isolated by contribution of Ficoll (1077) density gradient centrifugation. By using of trans well system, culture of tumor cell lines (MCF-7 , MDA-231) was performed adjacent with isolated cells. Lymphocyte proliferation assay by BrdU kit and immune-phenotyping of cytotoxic T lymphocyte(CD3+ CD8+) by Flow-cytometric analysis in two period of 48 and 72 h after co-culture was performed.
Results: Mean of lymphocyte proliferation was higher in multiparous women than nulliparous women and was statistically significant in 48 h and 72 h (P<0.001). Also, mean of lymphocyte proliferation of multiparous women in MCF-7 line (P=0.026) and in MDA-231cell line (P<0.001) showed significant results. Determination of the percentage of the population of cytotoxic lymphocytes (CD3 + CD8 +) showed no significant difference between two groups (P=0.603).
Conclusion: The cellular immune responses of multiparous mothers in co-culture with tumor cell lines have clear difference compared with nulliparous women that can lead to the resistance of the mothers against breast tumors. This pheromone could be related to micochimerism during pregnancy. Conclusion about the relation of breast tumor and pregnancy needs to investigate the anti-tumor responses with larger sample size.


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