Background. Human natural killer (NK) cells mediate spontaneous cytotoxicity against tumor cells and represent the main precursors of lymphokine‐activated killer (LAK) cell activity. A comparison of some aspects of NK and LAK cell activity was undertaken in 85 preoperative patients with breast cancer and 75 healthy donors. Methods. NK cell activity (tested in 18‐hour cultures of effector peripheral blood mononuclear cells [PBMC] with K562 or MOLT‐4 tumor target cells) was significantly diminished in these patients as it was the fully mature LAK cell activity (i.e., interleukin‐2 (IL‐2)‐in‐duced cytotoxicity in PBMC) against NK resistant target cells. Using immunoenzymatic methods we showed that the reduced NK cell activity was due to abnormally high levels of prostaglandin E2 (PGE2) produced by monocytes in culture. Results. PGE2 was found to suppress the production of IL‐2 in these cultures. Removal of monocytes from PBMC restored to almost normal levels the deficient NK and LAK cell activity in patients with breast cancer and was also associated with a normalization in the levels of PGE2 and IL‐2. Indomethacin and gamma‐interferon (IFN‐γ) increased the NK and LAK cell activity in these patients up to the levels of healthy donors. When highly purified CD56+ cells (obtained by an immunomagnetic isolation technique) were used as effector cells, no differences in LAK cell activity could be noticed between healthy donors and patients with cancer. FACS and northern blot analyses demonstrated a PGE2‐mediated down‐regulation of IL‐2 receptor (IL‐2R) expression on CD56+ cells that correlated with reduced LAK cell activity. This inhibitory effect of PGE2 was noticeable in long‐term LAK cultures and was abrogated in the presence of IFN‐γ or indomethacin. Conclusion. This study may have important implications in the potentiation of NK and LAK cell activity for immunotherapeutic protocols in patients with breast cancer.

Background. In vitro studies have demonstrated that exposure of tumor infiltrating lymphocytes (TIL) to human recombinant interleukin‐2 (rIL‐2) will generate activated T‐lymphocytes with major histocompatibility complex (MHC)‐restricted and non‐MHC‐restricted cytotoxicity toward a panel of tumor target cells. In melanoma and ovarian carcinoma, TII. display MHC‐restricted and autologous tumor‐specific cytotoxicity. Such tumor‐reactive cytotoxic T‐lymphocytes (CTL) represent important material for understanding cellular immunity in cancer and developing specific immunotherapeutic approaches in melanoma and ovarian cancer. In breast cancer, some TIL have been demonstrated to secrete cytokines upon interaction with autologous tumor cells, indicating that autologous tumor‐reactive lymphocytes may also exist among TIL in breast cancer. Therefore, the authors conducted a study to investigate the cytotoxic profile of rIL‐2‐activated lymphocytes in breast cancer. Methods. Lymphocytes were isolated from primary solid tumors (TIL) of breast carcinomas (10 patients) and from peritoneal effusions (effusion‐associated mononuclear cells [EAMNC]) from 2 patients with newly diagnosed metastatic breast carcinoma. Tumor infiltrating lymphocytes or EAMNC were cultured with rIL‐2 in long term cultures whereby their expansion index, phenotype, and cytotoxic potential were studied. Results. Both TIL and EAMNC proliferated by greater than 300‐fold (370‐3650; mean, 1656) after 23‐82 days in cultures containing mixtures of TIL or EAMNC, autologous tumor cells, and rIL‐2. By fluorescence analysis, freshly isolated TIL and EAMNC were found to consist of 77.5 plus or minus 10.7% CD3+ T‐cells, 33.2 plus or minus 8.9% CD4+, and 47.2 plus or minus 16.8% CD8+ cells. Their CD4 to CD8 ratio was 0.70. After expansion of lymphocytes with rIL‐2 in the majority of patients (9 of 12), CD3+CD8+ T‐lymphocytes were present in greater numbers than CD3+CD4+ T‐lymphocytes. Recombinant interleukin‐2‐activated CD3+CD8+ cells exhibited preferential cytolytic activity against autologous tumor cells. The cytolytic activity of CD3+CD8+ cells was inhibited either by anti‐T‐cell receptor (TCR)‐α/‐β and anti‐CD3 monoclonal antibodies (MoAb) or after pretreatment of tumor target cells with MoAb against the class I MHC antigens. Recombinant interleukin‐2‐activated CD3+CD4+ cells demonstrated potent cytolytic activity against both autologous and allogeneic tumor cells. CD3+CD8+ T‐cell clones isolated from representative TIL exhibited preferential autologous tumor‐specific cytotoxicity whereas the cytolytic activity of CD3+CD4+ T‐cell clones was mostly (12 of 14 clones) nonrestricted to the autologous tumor. Conclusions. To the authors' knowledge, this is the first report to demonstrate that TIL from primary tumors of breast carcinomas and EAMNC from metastatic disease can be propagated in large numbers in vitro with rIL‐2 while retaining autologous tumor specific and MHC‐restricted CTL activity. These findings may be of importance to ongoing clinical trials usi...

Breast cancer (BC) continues to affect the lives of millions of women worldwide. Several members of the carcinoembryonic antigen-related cell adhesion molecule (CEACAM) subfamily are involved in tumor progression. Notably, the CEACAM subfamily harbors the already established cancer biomarker CEA, as well as other potential molecular markers. CEACAM19, a recently identified gene belonging to CEACAM subfamily, was discovered and cloned by members of our research group. The present study analyzes, quantitatively, the expression of CEACAM19 and evaluates its clinical relevance in BC. Total RNA was extracted from 143 cancerous and 89 normal adjacent breast tissue specimens. Following reverse transcription, quantitative analysis of CEACAM19 mRNA expression levels was performed via real-time PCR and the comparative Ct (2-∆∆Ct) method. CEACAM19 expression and detailed clinicopathological data were used for extensive biostatistical analyses. CEACAM19 was found to be overexpressed in breast cancer tissue specimens compared to normal tissue counterparts (p=0.013). CEACAM19 mRNA expression status was also associated with clinicopathological features indicative of aggressive behavior and poor prognosis in BC, such as high tumor grade (p=0.031) and high Ki67 proliferative index (p=0.038). A significant negative association was documented between CEACAM19 expression and tumor ER status (p=0.018) as well as patients' menopausal state (p=0.016). Our results suggest that CEACAM19 mRNA expression represents a promising, novel and clinically useful tissue biomarker for breast cancer management.

The study and measurement of psychosocial adjustment is important for evaluating patients' well-being, and assessing the illness's course, treatment's success, and patients' recovery. In this study, internal consistency reliability and construct validity of the Greek version of the Psychosocial Adjustment to Illness Scale-Self-Report (PAIS-SR) were examined. Demographic and psychosocial data were collected from a sample of 243 women with breast cancer, recruited from September 2011 to December 2012. With some exceptions in specific items, the original conceptually-derived PAIS-SR subscales emerged in a seven-factor solution. Social Environment, Job and Household Duties, and Psychological Distress accounted for more of the total variance than other subscales. PAIS-SR showed good internal consistency reliability, with Cronbach's alpha coefficients >0.62. Correlations of PAIS-SR domains with measures of quality of life and posttraumatic stress symptoms supported the convergent validity of the PAIS-SR and its significance for cancer research. The Greek version of the PAIS-SR has acceptable internal consistency reliability and construct validity, as well as satisfactory convergent validity. Results provide some suggestions for the development of programs to evaluate adjustment status and implement psychosocial interventions among breast cancer survivors.

Kallikrein-related peptidases (KLKs) are a group of 15 serine proteases, hormonally regulated, and localized on chromosome 19q13.4. Alternative splicing is a process that plays significant role in the development, physiology, and different diseases, like cancer. Kallikrein family numbers more than 82 alternative transcripts. Understanding the role that those gene transcripts play in various cancer types, could lead to the discovery of diagnostic markers or drug targets. The present study was designed to analyze the expression profile of the splice variants of kallikrein-related peptidase 12 (KLK12) in breast cancer patients and to evaluate their clinical significance. KLK12 splice variants (KLK12sv3 and KLK12sv1/KLK12sv2) were examined in 69 tissue samples of breast cancer using quantitative real-time PCR as well as semi-quantitative PCR. Relative quantitative expression of KLK12 was statistically associated to clinicopathological parameters. From the splice variants examined, statistical associations with clinicopathological parameters were obtained only from KLK12sv3 variant. KLK12sv3 is more frequently expressed in tumors of lower grade (p = 0.040), early patient TNM stage (p = 0.024), and smaller tumor size (p = 0.023). Positive KLK12sv3 expression is associated with longer patient disease-free survival (DFS) (p = 0.042) and higher progesterone receptor concentration (p = 0.008). KLK12sv1/KLK12sv2 expression is statistically associated with KLK12sv3 expression (p = 0.001). KLK12sv3 can be regarded as a marker of good prognosis in breast cancer.