Checkpoint proteins regulate the immune system; breast cancer cells exploit the up-regulation or down-regulation of these proteins to evade anti-tumour immune responses . It is now well recognized that advanced metastatic BC and early disease are associated with both localized and systemic immune dysfunction .in this study levels of soluble immune checkpoint molecules sTIM3 and sCD40 will be measured and compared with tissue form ,then follow up to patients' prognosis and the relation to markers levels.

Breast cancer is the most common malignant tumour and the leading cause of cancer-associated mortality among women .Although comprehensive therapies exist, patient response to the treatments significantly varies, which partly attributed to varying antitumor immune responses .Immunotherapy is being recognized as a key therapeutic modality for cancer and represents one of the most promising therapies. Checkpoint proteins regulate the immune system. Breast cancer (BC) cells exploit the up-regulation or down-regulation of these proteins to evade anti-tumour immune responses. Notwithstanding the existence of profound immune dysregulation in advanced metastatic breast cancer (BC), it is now well recognized that early disease is also associated with both localized and systemic immune dysfunction . Recently, soluble co-inhibitory immune checkpoint molecules (ICMs) have been implicated as being potential mediators of the systemic immune dysregulation . Prominent among these soluble co inhibitory ICMs are: cytotoxic T-lymphocyte-associated protein (CTLA-4), programmed cell death protein 1 (PD-1) and its ligand, PD-L1, lymphocyte activation gene 3 (LAG-3) and T- Cell immunoglobulin and mucin-domain-containing protein 3 (TIM-3) . A study showed that resistance to anti-CTLA-4 or anti PD-1/PD-L1 inhibitors is compensated by up regulation of other immune checkpoints, such as Tim-3 . Consequently, Tim-3 has gained prominence as a potential candidate for cancer immunotherapy. Blocking Tim-3 with other checkpoint inhibitors has been shown to enhance antitumor immunity and suppress tumour growth in several preclinical tumour models. Increasing numbers of novel receptors and ligands have recently been found in the immune system. Some take part in a costimulatory interactions, such as Glucocorticoid-induced TNFR-related protein (GITR), GITR Ligand, CD27, CD28, CD40, CD80, CD86 and Inducible Co-Stimulator (ICOS) . CD40-CD40 ligand (CD40L) pathway is a member of the TNF superfamily and is expressed at various levels on antigen-presenting cells, epithelial cells, and hematopoietic progenitor cells . The CD40-CD40L costimulatory pathway has been shown to play a crucial role in humoral responses in humans, these cytokines modulate the function of T lymphocytes in antitumor responses. Few Review studies have reported the levels of sTIM-3 and CD40 in the literature, we are going to assess them and compare different stages of breast cancer .Previous study reported that, the level of Plasma concentration of the co-stimulatory immune checkpoint CD40 as well as the co-inhibitory molecule TIM-3 were all significantly lower in early breast cancer patients compared to healthy controls. Following neoadjuvant chemotherapy NAC, the plasma concentrations of the soluble co-stimulatory ICM sCD40 and soluble co inhibitory ICM sTIM-3 were significantly increased

Inclusion Criteria:

• Female patients with breast cancer aged > 18 years attending the Medical Oncology department of South Egypt Cancer Institute

Exclusion Criteria:

• Patients on chemotherapy
• A history of any other malignancy
• known (HIV) and/or hepatitis B or hepatitis C viruses,
• pregnancy or breast feeding
• patients or controls that refuse to be a part of the study.

Female patients with breast cancer and healthy females as controls