Breast cancer immunotherapy: Current and novel approaches.

The crucial role of the immune system in the progression/regression of breast cancer (BC) should always be taken into account. Various immunotherapy approaches have been investigated for BC, including tumor-targeting antibodies (bispecific antibodies), adoptive T cell therapy, vaccines, and immune checkpoint blockade such as anti-PD-1. In addition, a combination of conventional chemotherapy and immunotherapy approaches contributes to improving patients' overall survival rates. Although encouraging outcomes have been reported in most clinical trials of immunotherapy, some obstacles should still be resolved in this regard. Recently, personalized immunotherapy has been proposed as a potential complementary medicine with immunotherapy and chemotherapy for overcoming BC. Accordingly, this review discusses the brief association of these methods and future directions in BC immunotherapy.

Breast Tumor Cells Highly Resistant to Drugs Are Controlled Only by the Immune Response Induced in an Immunocompetent Mouse Model.

BACKGROUND: The tumor cells responsible for metastasis are highly resistant to chemotherapy and have characteristics of stem cells, with a high capacity for self-regeneration and the use of detoxifying mechanisms that participate in drug resistance. In vivo models of highly resistant cells allow us to evaluate the real impact of the immune response in the control of cancer. MATERIALS AND METHODS: A tumor population derived from the 4T1 breast cancer cell line that was stable in vitro and highly aggressive in vivo was obtained, characterized, and determined to exhibit cancer stem cell (CSC) phenotypes (CD44+, CD24+, ALDH+, Oct4+, Nanog+, Sox2+, and high self-renewal capacity). Orthotopic transplantation of these cells allowed us to evaluate their in vivo susceptibility to chemo and immune responses induced after vaccination. RESULTS: The immune response induced after vaccination with tumor cells treated with doxorubicin decreased the formation of tumors and macrometastasis in this model, which allowed us to confirm the immune response relevance in the control of highly chemotherapy-resistant ALDH+ CSCs in an aggressive tumor model in immunocompetent animals. CONCLUSIONS: The antitumor immune response was the main element capable of controlling tumor progression as well as metastasis in a highly chemotherapy-resistant aggressive breast cancer model.

Characterization and antitumor activities of the water-soluble polysaccharide from Rhizoma Arisaematis.

A water soluble polysaccharide (RAP-W1) was purified from Rhizoma Arisaematis and its antitumor activity was evaluated in BALB/c mice bearing human breast cancer MCF-7. RAP-W1 had the following physicochemical properties: total carbohydrate content (95.9%); no protein; molecular weight (≈57 kDA); monosaccharides composition (rhamnose:fucose:arabinose:mannose:galactose:glucose=0.4:0.5:0.3:0.6:0.9:5.3). After 14 days' treatment to tumor-bearing mice, RAP-W1 could significantly inhibit the growth of tumor transplanted in mice and increase the body weight and the spleen index. Moreover, RAP-W1 could significantly stimulate Con A- or LPS-induced splenocyte proliferation in tumor-bearing mice, as well as enhance the CTL activity. The level of Th1 cytokines (INF-γ and IL-2) in the serum of tumor-bearing mice was increased by RAP-W1 treatment, whereas the Th2 cytokine (IL-10) secretion displayed a dramatic reduction. All the data implied that RAP-W1 can activate T cells by up-regulating Th1/Th2 cytokine ratio, which might partially cause the inhibition of tumor growth.

Detection of interferon-gamma-inducible chemokines in human milk.

AIM: To assess the immunological role of human milk by analysing the concentrations of interferon-gamma-inducible protein of 10 kda (IP-10) and monokine induced by interferon-gamma (MIG) in human milk from mothers of preterm and term infants. METHODS: IP-10 and MIG levels of colostrum, early milk, mature milk and sera were measured by enzyme-linked immunosorbent assay (ELISA). IP-10 and MIG mRNA expression levels in cellular components of human milk were determined by RT-PCR. IP-10 and MIG protein expression in mammary gland tissues was analysed by immunohistochemistry. RESULTS: Significant amounts of IP-10 and MIG were detected in human milk. The concentrations of IP-10 and MIG in colostrum and early milk were significantly higher than those of sera from healthy controls or lactating mothers. These chemokine concentrations in colostrum and early milk were significantly higher than those of mature milk. Premature delivery or pregnancy complications of mothers had no significant correlation with these chemokine concentrations in breast milk. There were significant correlations between MIG and interferon-gamma (IFN-gamma) or IP-10 levels (p < 0.001) in human milk. Expression of IP-10 and MIG genes and proteins in the milk cells as well as in mammary gland epithelial tissues was detected by RT-PCR and immunohistochemistry. CONCLUSION: IP-10 and MIG in human milk, probably derived from milk cells and mammary gland epithelial cells, may contribute to the migration and activation of intestinal T lymphocytes to enhance mucosal immunity during the early neonatal period.

Interleukin-18 in human milk.

We analyzed IL-18 levels of human milk. Colostrum contained significantly higher levels of IL-18 compared with early milk and mature milk. By stepwise multiple linear regression analysis, preterm delivery and pregnancy complications of mothers significantly correlated with high levels of IL-18 in human milk (p = 0.0007 and 0.0018, respectively). There was a significant correlation between the levels of IL-18 and soluble Fas ligand in colostrum (p = 0.0003). IL-18 was detected in actively secreting epithelial cells in lactating mammary gland by immunohistochemical staining. These results suggest that IL-18 in colostrum plays an important role in host defense of high-risk neonates.

Identification of the complement regulatory protein CD59 in human colostrum and milk.

PROBLEM: Complement lytic activity has been demonstrated, and a potential for its activation is present in human colostrum and milk. This necessitates the presence of regulatory mechanisms protecting epithelial cells in the oropharynx and the gastrointestinal tract of the infant, the milk cellular elements, and bacteria colonizing the oropharynx and the gastrointestinal tract. Lactoferrin and C1 inhibitor have been attributed such a role. However, it is likely that additional protection against the cytolytic activity of the membrane attack complex is required. This has lead us to investigate the presence of the complement regulatory protein CD59 in human colostrum and milk, and to further characterize the source of secretion. METHOD: Samples of human colostrum and milk were obtained from volunteers at different stages of lactation, and separated into fat, skim milk, and milk cellular elements by centrifugation. Normal human mammary gland tissues were obtained from patients undergoing biopsy for benign conditions. SDS-PAGE and Western blotting, and an immuno dot-blot assay were used to identify CD59 in human milk. Immunohistochemistry was performed on all tissue samples and cytospins of the milk cellular elements, using monoclonal antibodies to CD59. RESULTS: CD59 was present in cell-free colostrum and milk as a 19-25 kDa glycoprotein. No variation in CD59 levels was detected between colostrum and milk. CD59 was present in great amounts in the cytoplasm and was highly expressed on the surface membrane on mammary gland acinar and ductal epithelial cells, while the milk cellular elements contained CD59 mainly in their cytoplasm. CONCLUSION: The complement regulatory protein CD59 present in cell-free human colostrum and milk may exert its effects both in the mammary gland and in the oropharynx and gastrointestinal tract of the infant. The lobuloalveolar epithelial cells in the mammary gland are the likely source of secretion.

Distribution of CA 125 in embryonic tissues and adult derivatives of the fetal periderm.

New murine monoclonal antibodies to a partially purified CA 125 antigen were developed and identified as M 2 and M 11. With immunohistochemical techniques, these new antibodies and OC 125 antibody were used to search for CA 125 in embryonic tissues and adult apocrine sweat glands and mammary glands. The embryonic skin, the periderm, expressed CA 125 antigen and its adult derivatives, the mammary glands and apocrine sweat glands, expressed CA 125 while in the active state of secretion. In a 6-week-old formalin-fixed and paraffin-embedded ectopic embryo specimen, antibodies M 2 and M 11 recognized CA 125 in the periderm, the notochord, the myocardium, the pericardium, the gastroenteric tract, enteric duct remnants in the umbilical cord (vitelline and allantoic ducts), the mesonephric duct, and the amnion. OC 125 staining of these formalin-fixed specimens was either very faint or absent. In a formalin-fixed and paraffin-embedded specimen of apocrine sweat glands from the axilla, antibodies M 2 and M 11 detected CA 125 antigen intracellularly in the secretory cells. Again no staining was observed with OC 125 antibody. In a frozen and acetone-fixed specimen of lactating mammary glands, antibodies M 2 and OC 125 detected CA 125 antigen intraductally. Colostrum and milk collected from 25 mothers at various stages post partum had mean CA 125 levels of 34,213 U/ml in colostrum, 1469 U/ml at 3 to 7 days, and 105 U/ml at 5 to 26 weeks.

Secretion and composition of colostrum and milk.

Lacteal fluids contain antibodies of immunoglobulin (Ig) A, IgG, and IgM classes, and these proteins are complete and biologically active. Lymphocytic cells have also been demonstrated that are capable of an array of biological activity characteristic of similar cells in the blood. Lacteal fluids are rich in phagocytic cells that function less efficiently than do their corresponding blood cells.

Maternal-neonatal interactions and human breast milk.

The mammary glands are a uniquely designed extension of the mucosal immune systems of the gut and bronchus. The soluble and cellular products of lactation, in which immunologic selectivity and specificity exist, link the suckling neonate irrevocably to the immunologic and infectious experience of its mother. The immune competence of the breast and products of lactation are actively and constantly in flux, dependent on the mother's hormonal, environmental, and immunologic milieu. Most of the recent human investigation has focused primarily on milk immunoglobulin specificity. More work needs to be done on the role of passively transferred cellular products, the antiviral, antiprotozoan, and antitumor capabilities of milk, and on the mechanisms by which maternal immunizations or infections may influence the outcome of host-pathogen interactions in the suckling neonate.

Immunological functions of the mammary gland and its secretion--comparative review.

The mammary gland performs vitally important immunological roles, both in providing passive immune protection to the suckling infant and in immunological defence of its own tissues against infection with microorganisms. These immunological functions differ greatly between species of mammals in both nature and magnitude. In ungulates the mammary gland is singularly responsible for transfer of immunoglobulin (IgG) from mother to young. This process is dependent on a highly selective mechanism which results in the transport of blood-borne IgG molecules across secretory epithelial cells of the colostrum-forming mammary gland and into secretion. Upon ingestion of colostrum by the young ungulate this immunoglobulin is absorbed across the wall of the small intestine and thence into the bloodstream. In other species, including rodents and primates, there is a well-developed local IgA system operating in the mammary gland. In this situation, plasma cells located near the basal membranes of secretory epithelial cells secrete IgA which passes through the epithelial cells and into colostrum of milk. In the species the IgA in mammary secretions is not absorbed into the circulation of the suckling infant; because of its unique property of resisting proteolytic degradation, it may mediate a local protective role in the lumen of the intestine of the suckling infant. Specific immunological protection of mammary tissue may be mediated through blood-derived antibody (particularly IgG), locally synthesized antibody (particularly IgA) or phagocytic cells. Neutrophils arrive in mammary tissue and secretions in very large numbers following bacterial invasion of the gland. It has been established recently that these cells carry cytophilic antibody on their cell membrane. This cytophilic antibody can play an important functional role in enhancing the phagocytic capacity of neutrophils in the mammary gland.