CD8-positive memory T cells in tumor-draining lymph nodes of patients with breast cancer.

BACKGROUND: Human immunological memory is a hallmark of the adaptive immune system and plays an important role in the development of effective immune responses against tumors. In the present study, we aimed to determine the frequencies of CD8+ memory T cell subsets including T stem cell memory (TSCM) in tumor-draining lymph nodes of patients with breast cancer (BC). METHODS: Mononuclear cells were obtained from axillary lymph nodes of 52 untreated patients with BC and stained for CD8, CCR7, CD45RO, CD95 markers to detect different subtypes of memory cells in the CD8+ lymphocyte population. Data were acquired on four-color flow cytometer and analyzed with CellQuest Pro software. RESULTS: We observed that 47.65 ± 2.66% of CD8+ lymphocytes expressed the CD45RO, a marker for memory T cells. Statistical analysis showed that the total frequency of central memory T cells (TCM) and their subset with low CD45RO expression was significantly higher in tumor-involved nodes compared to tumor-free ones (P = 0.024 and P = 0.017, respectively). The level of CD95 expression (based on mean fluorescence intensity) on the surface of TCM, their CD45ROhi and CD45ROlow subsets, and TSCM was higher in patients with stage II compared to those in stage I (P < 0.05). In addition, the percentage of naive CD8+ T cells was significantly lower in tumor-involved lymph nodes compared to tumor-free ones (P = 0.025). CONCLUSIONS: Our data collectively indicate no significant differences in the frequencies of CD8+ lymphocytes or their memory subsets in tumor-draining lymph nodes of patients with BC. However, the frequency of CD45low TCM was higher in tumor-involved nodes. Along with a decrease in the frequency of naive T cells, the higher frequency of CD45low TCM suggests that despite the immune reaction to provide a pool of effective memory cells, it is blocked in early-stage of memory cells' differentiation (CD45ROlow), probably by tumor-derived suppressive factors. Identifying the molecular and cellular mechanisms behind this suppression can provide invaluable tools for adoptive T cell therapies in cancer.

Mesenchymal stem cells induced anti-inflammatory features in B cells from breast tumor draining lymph nodes.

The immune-modulatory effect of adipose-derived stem cells (ASCs) on B cells in cancer has not been well elucidated. Herein, the interaction between B cells and ASCs isolated from the breast fat of either normal (nASCs) or breast cancer women (cASCs) was investigated. B cells derived from breast tumor draining lymph nodes were co-cultured with nASCs or cASCs and B cells proliferation was assessed in direct and transwell assays. Moreover, B cells were co-cultured with cASCs, nASCs or mesenchymal stromal cells of the tumor tissue (TSCs) and B cell cytokine production was assessed using flow cytometery. cASCs or TSCs were co-cultured with either intact or B cell depleted lymphocytes and frequencies of CD25+ FoxP3+ Tregs, IL-10+ or IFN-γ+ CD4+ T cells were assessed. Results showed that co-culture of B cells with ASCs in transwell chambers did not affect B cell proliferation. nASCs, however, was able to significantly reduce B cell proliferation in direct co-culture experiments (P = 0.004). The frequencies of IL-10+ , TNF-α+ , IL-2+ , and IFN-γ+ B cells were not significantly different in the co-cultures of B cells with ASCs or TSCs. But the TNF-α+ / IL-10+ B cells ratio decreased in all co-cultures, a reduction merely significant in B cell-cASCs co-culture (P = 0.01). The frequencies of CD4+ T cells subsets in either intact or B cell depleted lymphocytes did not undergo significant changes following co-culture with ASCs or TSCs. Therefore, ASCs is capable of inhibiting B cell proliferation in a contact dependent manner and shifting the cytokine profile of B cells toward an anti-inflammatory profile.

Langerhans cell histiocytosis followed by hodgkin lymphoma: a case report.

Langerhans cell histiocytosis (LCH) is a rare neoplasm defined as the proliferation of bone marrow langerhans cells, which is a kind of dendritic cells. The major pathological features of LCH are expression of CD1a and S100 as well as Birbeck granules. Its presentation can differ from a mild bone lesion to a multi-systemic evolved malignant neoplasm; however, the latter outcome is almost rare. Thus, LCH is mostly known as a benign neoplasm. In this study, we present a case of LCH followed by Hodgkin lymphoma (HL). Accompaniment of this disease with malignant lymphoma is rare and considered as case report. Several cases in which malignant lymphoma occurred prior to LCH are reported; however, few cases can be found with LCH followed by malignant lymphomas.

Extragonadal sclerosing stromal tumor: a rare case report.

Sclerosing stromal tumor is a rare, benign, sex cord stromal tumor of the ovary. We report a case of extragonadal sclerosing stromal tumor in a 45-year-old woman who presented with menstrual irregularity and vague pelvic pain. Imaging studies showed a well-defined mass between the posterior wall of the bladder and uterus, suspected of being a pedunculated leiomyoma. The histopathological and immunohistochemical study was consistent with sclerosing stromal tumor. No ovarian tissue was found on representative sectioning. This is the first case of sclerosing stromal tumor in an extragonadal location.

Analysis of ß/α globin ratio by using relative qRT-PCR for diagnosis of beta-thalassemia carriers.

BACKGROUND: Current routine tests for premarital screening of ß-thalassemia carriers are not applicable for diagnosis of rare atypical minor ß-thalassemia cases. A more specialized laboratory evaluation for them is the measurement of ß/α chain synthesis ratio with the assistance of radioactive amino acids. This method is also no longer routinely accessible. Consequently it is required to establish a rapid, trouble-free, and reliable method that encompasses all the cases of ß-thalassemia carriers. Therefore we have determined ß/α-globin mRNA ratio by applying relative qRT-PCR in various ß-thalassemia patients. METHODS: Reticulocytes RNA extraction and subsequent cDNA synthesis were performed, followed by relative qRT-PCR for α- and ß-globin chain genes and ß-actin gene as an endogenous reference. ß/α-Globin gene ratio was then evaluated with the Pfaffl method. RESULTS: The mean of ß/α ratio was 0.99, 0.81, 0.69, and 0.69 for normal population, minor, intermediate, and major ß-thalassemia, respectively. Approximately 6% of cases with minor thalassemia RBC index and normal HbA2 and having a decreased ß/α ratio were located in the minor ß-thalassemia group. The mean of ß/α mRNA ratio in normal individuals and minor ß-thalassemia was significantly different with all other groups (P-value < 0.05). Nevertheless, there was no such association between ß/α mRNA ratio in major and intermediate ß-thalassemia. CONCLUSION: According to the significant differences achieved, no overlapping between minor ß-thalassemia and normal group, capability of diagnosing atypical minor ß-thalassemia, and accessibility of this technique, we can declare that this method could be suggested as a routine premarital screening test for ß-thalassemia carriers.

The significance of cytokine-producing B cells in breast tumor-draining lymph nodes.

PURPOSE: The role of cytokine-producing B cells in antitumor immunity is mostly overlooked. In the present study, we investigated changes in B cell cytokine profiles in breast tumor-draining lymph nodes (TDLNs) during disease progression, and associations of these changes with prognostic indicators. METHODS: Flow cytometry was used to measure the expression of TNF-α, IL-10, TGF-ß, IL-2 and IFN-γ in B cells from 42 axillary lymph nodes. The frequencies of IL-10+ and FoxP3+ T regulatory cells (Tregs) were also determined. RESULTS: No significant changes in B cell cytokine profiles were observed during breast cancer progression from stage I to III, but the percentage of B cells with high TNF-α expression (TNFhi) showed a negative relationship with lymph node involvement and Her2 expression (p < 0.05). The percentage of IL-10+ B cells was found to be significantly higher in non-metastatic lymph nodes in node-negative compared to node-positive patients (p = 0.001). The frequencies of IL-10+ and TNFhi B cells were found to be negatively correlated with the number of involved lymph nodes. The frequency of TNFhi B cells showed an inverse correlation with the frequency of FoxP3+ Tregs, which in turn was associated with indicators of a poor prognosis. CONCLUSIONS: Our data indicate that the cytokine profiles of B cells in TDLNs of patients with breast cancer show associations with various disease parameters. TNFhi and IL-10+ B cells correlated positively with indicators of a good prognosis. Further functional studies are required to elucidate the role of cytokine production by B cells in immunity against breast cancer.

Expression of maspin and ezrin proteins in periocular Basal cell carcinoma.

Background. The aim of this study was to investigate maspin and ezrin expression in different subtypes of periocular basal cell carcinoma (BCC). Methods. Tissue samples from 43 patients with periocular BCC. Our cases were comprised of 10 morpheaform, 25 nodular, and 8 adenoid type BCCs. Immunohistochemical staining for maspin and ezrin was performed by Envision detection system. Results. There was no difference between different subtypes of BCC in maspin expression regarding positivity, intensity, and pattern of expression. Ezrin was expressed in all subtypes of BCC but the intensity was significantly higher in morpheaform BCC compared to nodular and adenoid types (P < 0.001 and P = 0.012, resp.); ninety percent of morpheaform samples showed strong ezrin intensity, while this strong intensity was only present in 25% and 12% of adenoid and nodular subtypes, respectively. There was no correlation between age, sex, or tumor margin involvement and expression of neither maspin nor ezrin. There was no correlation between maspin and ezrin expression except in nodular type, in which an inverse correlation was found (P = 0.004). Conclusion. Ezrin is expressed intensely in morpheaform BCC of periocular region. Further studies are needed to show the significance of this finding in prognosis of morpheaform BCC.

Relationship between AHSP gene expression, ß/α globin mRNA ratio, and clinical severity of the ß-thalassemia patients.

BACKGROUND: Alpha hemoglobin stabilizing protein (AHSP) is a chaperone-like molecule specialized for erythroid series which binds to free α-globin chain. According to this characteristic, AHSP can be considered an important factor which reduces beta thalassemia symptoms. MATERIALS AND METHODS: Reticulocytes RNA extraction and a subsequent cDNA synthesis were performed, followed by Relative qRT-PCR for AHSP, alpha, and beta globin chain genes. The beta actin gene was used as an endogenous reference as well. The relationship between AHSP gene expression, disease severity, and the ß/α globin mRNA ratio was studied among different homozygote ß-thalassemia patients (mild, moderate and severe) and compared with minor thalassemia and the normal population. RESULTS: Analysis of the ß-globin/α-globin mRNA ratio has shown that disease severity enhanced with a decrease in this proportion. Evaluation of the correlation between AHSP gene expression and the average of the ß-globin/α-globin expression ratio indicated a significant but indirect relationship in considered groups. Our results demonstrated that the AHSP gene expression increases in accordance with augmentation of clinical symptoms. CONCLUSIONS: Although one of the main reasons for reduced clinical severity in homozygote ß-thalassemia can be the high level of AHSP gene expression as a chaperon molecule, our findings indicated that AHSP gene expression decreased in a mild category as compared to that in severe and moderate groups.

A pilot study on lipolytic effect of subcutaneous botulinum toxin injection in rabbits.

OBJECTIVE: To determine whether botulinum toxin type A (BTX-A) exerts a lipolytic effect by interfering with acetylcholine transmission at the cholinergic parasympathetic nerve endings. STUDY DESIGN: Fifteen male rabbits were divided into 3 equal groups: 1 control group (A) and 2 case groups (B and C). The abdomens of all rabbits were divided into a 3 x 3-square grid. The groups received 9 subcutaneous injections of 0.9% normal saline, 1 U BTX-A (group B) and 2 U BTX-A (group C), respectively. Four weeks later the entire grid was excised from the abdominal area. Hematoxylin-eosin-stained tissue was used for stereologic analysis to estimate cell surface and volume in 100 randomly selected cells. RESULTS: Gross thinning of subcutaneous fat and shattering and disappearance of fat globules were seen in both case groups. Fat cell volume was reduced by 65% in group B (p = 0.009) and 77% in group C (p = 0.009) compared to control animals. Fat cell surface also decreased by 51% in group B (p = 0.009) and 63% in group C rabbits (p = 0.009) compared to control animals. CONCLUSION: Our pilot animal study revealed a dose-dependent lipolytic effect of subcutaneous BTX-A injection.