Regulatory T cells in allogeneic hematopoietic stem cell transplantation: From the lab to the clinic.

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a curable strategy for the treatment of hematological malignancies and nonmalignant diseases. However, graft-versus-host disease (GVHD) and relapse are still two major causes of morbidity and mortality after allo-HSCT, and both restrict the improvement of transplant outcomes. Regulatory T cells (Tregs) has been successfully used in allo-SCT settings. In this review, we summarize recent advances in experimental studies that have evaluated the roles played by Tregs in the establishment of novel transplant modalities, the prevention of GVHD and the enhancement of immune reconstitution. We also discuss the application of Tregs in clinical to prevent acute GVHD, treat chronic GVHD, as well as enhance immune reconstitution and decrease leukemia relapse, all of which lead to improving transplant outcomes.

Association of increased microvessel density with skeletal extramedullary disease relapse in multiple myeloma patients who have skeletal extramedullary disease at diagnosis.

The aim of the study was to investigate whether microvessel density (MVD) could be associated with skeletal extramedullary disease relapse (skeletal-EMDR) in patients with multiple myeloma (MM) who have skeletal-EMD at diagnosis. Seventy-nine newly diagnosed MM patients who have skeletal-EMD were retrospectively enrolled in this study. The 4-year cumulative incidence of skeletal-EMDR was 35.0%±8.3%. The 4-year probability of overall survival (OS) was 54.0%±7.6%. Multivariate analysis showed that skeletal-EMDR (HR = 4.144; 95% CI: 1.608-10.685; P = 0.003) was independently associated with inferior OS for the MM patients who have skeletal-EMD at diagnosis. The factors associated with skeletal-EMDR were MVD (HR = 3.990, 95%CI:1.136-14.018; P = 0.031), white blood cell (WBC) (HR = 0.262, 95% CI:0.090-0.769; P = 0.015), and the EMD sites involved at onset (HR = 0.263, 95% CI: 0.074-0.937; P = 0.039). The MVD in patients with thoracic and lumbar vertebrae as the involved sites at diagnosis was significantly lower than those with other sites involved (41.59 ± 14.39 vs. 60.82 ± 35.14, P=0.001). Our data suggest that increased MVD could be used to predict skeletal-EMDR, which is associated with inferior survival in patients with MM who have skeletal-EMD at diagnosis.

Correlation of graft immune composition with outcomes after allogeneic stem cell transplantation: Moving towards a perfect transplant.

Allogeneic stem cell transplantation (allo-SCT) offers an important curative therapy for hematological malignancies and other diseases. A number of studies have demonstrated the association of immune compositions in allografts with outcomes after allo-SCT, which promote graft engineering to improve transplant prognosis. This review summarizes the advances in investigating the correlation of the graft immune compositions with transplant outcomes in different transplant modalities, focusing on the immune subsets likely to have the greatest impact on clinical outcomes. The progress made in graft engineering in order to design novel transplant protocols, to decrease graft-versus-host disease and relapse and to improve immune recovery is also discussed. It is our belief that an adoptive immune subset transfer to improve clinical outcomes might represent a future direction.

High nuclear expression of Twist1 in the skeletal extramedullary disease of myeloma patients predicts inferior survival.

The aim of the study was to investigate the expression of epithelial to mesenchymal transition (EMT)-inducing transcription factors, including Twist1 and ZEB1, in skeletal extramedullary disease (EMD) of multiple myeloma (MM) patients and to clarify the effects on clinical outcomes. The expression of Twist1 and ZEB1 in the bone marrow (BM) and the masses of skeletal EMD from 70 MM cases with skeletal EMD and 30 MM patients without skeletal EMD were determined by immunohistochemistry. The results demonstrated that the percentage of high nuclear staining for Twist1 was 24.3% (17/70) in skeletal EMD, which was significantly higher than in the BM of these patients as well as those without skeletal EMD (P=0.030 and P=0.011). The microvessel density (MVD, P=0.004) was significantly higher in patients with high nuclear expression of Twist1 (Twist1-high) than in those with low expression. Patients with Twist1-high experienced a lower rate of progression-free survival (PFS, 11.8% vs. 35.0%, P=0.000) and overall survival (OS, 52.5% vs. 83.7%, P=0.001) compared to those with low expression. Multivariate analysis showed that Twist1-high was independently associated with inferior PFS (HR=2.161; 95%CI: 1.116-4.183; P=0.022) and OS (HR=3.111; 95%CI: 1.114-8.685; P=0.030). We concluded that Twist1-high is associated with a poor prognosis and may be correlated with angiogenesis in the skeletal EMD of MM patients.

Mechanisms for T cell tolerance induced with granulocyte colony-stimulating factor.

Granulocyte colony-stimulating factor (G-CSF) has been widely accepted as a mediator of T cell tolerance. The immune modulatory effect of G-CSF on T cells is believed to be mediated exclusively through other effector cells, such as monocytes, tolerogenic dendritic cells (DC), and myeloid-derived suppressor cells. Recent advances confirmed the direct effects of G-CSF in inducing immune tolerance of T cells through the G-CSF-G-CSF receptor pathway and related molecular mechanisms. This review aims to summarize the findings associated with the direct and indirect mechanisms for T cell tolerance induced with G-CSF. The role of G-CSF in preventing graft-versus-host disease (GVHD) and in treating autoimmune diseases (ADs) is also discussed. It is conceivable that G-CSF and immune cell compositions, such as tolerogenic DC and CD4(+)CD25(+)Foxp3(+) T cells, modulated by G-CSF could become an integral part of the immunomodulatory therapies against GVHD and ADs in the future.

Nimesulide inhibits the growth of human esophageal carcinoma cells by inactivating the JAK2/STAT3 pathway.

Although selective COX-2 inhibitors have cancer-preventive effects and induce apoptosis, the mechanisms underlying these effects are not fully understood. This study investigated the effects of nimesulide, a selective COX-2 inhibitor, on apoptosis and on the JAK/STAT signaling pathway in Eca-109 human esophageal squamous carcinoma cells. The effects and mechanisms of nimesulide on Eca-109 cell growth were studied in culture and in nude mice with Eca-109 xenografts. Cells were cultured with or without nimesulide and/or the JAK2 inhibitor AG490. Cell proliferation was evaluated using the MTT assay, and apoptosis was investigated. COX-2 mRNA expression was measured using reverse transcription polymerase chain reaction, and protein expression was detected by Western blot analysis, immunohistochemistry, and flow cytometry. Nimesulide significantly inhibited Eca-109 cell viability in vitro in a dose- and time-dependent manner (P<0.05). Nimesulide also induced apoptosis, which was accompanied by a significant decrease in the expression of COX-2 and survivin and an increase in caspase-3 expression. Nimesulide downregulated the phosphorylation levels of JAK2 and STAT3, and JAK2 inhibition by AG490 significantly augmented both nimesulide-induced apoptosis and the downregulation of COX-2 and survivin (P<0.05). In vivo, nimesulide inhibited the growth of Eca-109 tumors and the expression of p-JAK2 and p-STAT3. Thus, nimesulide downregulates COX-2 and survivin expression and upregulates caspase-3 expression in Eca-109 cells, by inactivating the JAK2/STAT3 pathway. These effects may mediate nimesulide-induced apoptosis and growth inhibition in Eca-109 cells in vitro and in vivo.

Interleukin-17 receptor expression on vascular endothelial cells of masses of skeletal extramedullary disease in myeloma patients.

The goal of the study was to investigate the expression of interleukin-17 (IL-17) and IL-17 receptor (IL-17R) in patients with myeloma bone diseases (MBD) and skeletal extramedullary disease (skeletal EMD). The levels of IL-17 were determined using ELISA. The expression of IL-17R on vascular endothelial cells of bone marrow (BM) and masses of skeletal EMD was detected using immunohistochemistry. The results showed an elevated IL-17 level in BM of BMD and skeletal EMD patients. The microvessel density (MVD) was significantly increased in the masses of skeletal EMD. IL-17R was almost exclusively expressed by endothelial cells, not by myeloma cells in the masses of skeletal EMD patients. We concluded that EMD masses showed increased angiogenesis mediated by IL-17 pathway and in part this may help in myeloma cell-growth under these conditions.

Expression levels of IL-27 and IL-17 in multiple myeloma patients: a higher ratio of IL-27:IL-17 in bone marrow was associated with a superior progression-free survival.

The goal of the study was to investigate the levels of interleukin-27 (IL-27) and IL-17 in bone marrow (BM) and peripheral blood (PB) of multiple myeloma (MM). The levels of IL-27 and IL-17 were determined in MM patients and controls using ELISA. The results showed a decreased IL-27 and elevated IL-17 level in MM patients and a negative association of IL-27 with IL-17. The ratio of IL-27:IL-17 in BM of newly diagnosed MM was significantly decreased and correlated with the progression of disease. Multivariate analysis showed that a higher ratio of IL-27:IL-17 in BM was associated with a superior progression-free survival (HR=0.160; 95% CI: 0.058-0.443; p<0.001). Our results suggest that there might be a possible competitive role of IL-27 and IL-17 in MM.

Decreased soluble TGF-ß1, Tie-2, and angiopoietins serum levels in bone marrow after treating healthy donors with granulocyte colony-stimulating factor.

The goal of this study was to investigate the effect of granulocyte colony-stimulating factor (G-CSF) on Tie-2, angiopoietins, VEGF, and TGF-ß1 in bone marrow (BM) of healthy donors. Soluble Tie-2, angiopoietins, VEGF, and TGF-ß1 levels in the BM were determined via ELISA in 25 healthy donors before and after G-CSF treatment. The results showed that treating healthy donors with G-CSF significantly decrease serum levels of Tie-2, angiopoietin-1, angiopoietin-2, and TGF-ß1. In contrast, median VEGF level in the G-CSF-primed BM was significantly higher than steady-state BM. Our results suggest that decreased soluble TGF-ß1, Tie-2, and angiopoietins levels in the BM could be related to stem cell mobilization.

[Expression and significance of N-cadherin and ß-catenin protein in osteosarcoma].

OBJECTIVE: To evaluate the expression of N-cadherin and ß-catenin protein and their relationship with clinicopathological characteristics of osteosarcoma. METHODS: The expressions of N-cadherin and ß-catenin at protein level were detected by immunohistochemical staining in 54 cases of osteosarcoma, 11 cases of osteoid osteoma, 7 cases of osteoblastoma and 8 cases of newly formed bone in nonmalignant bone diseases. The relationship between the two indexes and clinicopathological characteristics of osteosarcoma was analyzed. RESULTS: In newly formed bone, osteoblastoma, osteoid osteoma and osteosarcoma, the positive expression rate of N-cadherin protein was 75.0%, 71.4%, 63.6% and 35.2%, respectively. The positive expression rate of N-cadherin protein in osteosarcoma was significantly lower than that in osteoid osteoma, osteoblastoma and newly formed bone in nonmalignant bone diseases (P = 0.035). The positive expression rate of N-cadherin protein in osteosarcoma cases with pulmonary metastasis was lower than that in cases without (21.7% vs. 56.3%, P = 0.027). The positive expression rate of N-cadherin protein in osteosarcoma cases died in two years was lower than that in cases lived for more than two years (18.2% vs. 50.0%, P = 0.024). In newly formed bone, osteoblastoma, osteoid osteoma and osteosarcoma, the aberrant expression rate of ß-catenin protein was 12.5%, 28.6%, 27.3% and 66.7%, respectively. The aberrant expression rate of ß-catenin protein in osteosarcoma was significantly higher than that in osteoid osteoma, osteoblastoma and newly formed bone (P = 0.002). Aberrant expression rate of ß-catenin in osteosarcoma cases with pulmonary metastasis was higher than that without (82.6% vs. 43.8%, P = 0.011). An inverse correlation was found between the aberrant expression of ß-catenin and N-cadherin expression in osteosarcoma(r = -0.302, P = 0.027). CONCLUSION: The positive expression rate of N-cadherin is decreased in osteosarcoma while aberrant expression rate of ß-catenin increased. The expression of N-cadherin protein is closely correlated with the metastasis and prognosis of osteosarcoma, but the expression of ß-catenin protein is merely correlated with the metastasis of osteosarcoma.

[Expression of EP-CAM, beta-catenin in the carcinogenesis of squamous cell carcinoma of uterine cervix].

OBJECTIVE: To study the expression of EP-CAM, beta-catenin in the carcinogenesis of squamous cell carcinoma of uterine cervix. METHODS: The expressions of EP-CAM and beta-catenin were detected with immunohistochemical stain in 14 cases of normal cervical squamous epithelium, 32 cases of cervical intraepithelial neoplasia (CIN) and 38 cases of cervical invasive squamous cell carcinoma. RESULTS: The over-expression rates of EP-CAM were 0, 7.1%, 20.0%, 62.5% and 55.3% for normal cervical epithelium, CINI, CINII, CINIII and carcinoma groups. The EP-CAM over-expression rates in CINIII and cervical carcinoma groups were significantly higher than those in normal epithelium and CINI groups (P < 0.001). No aberrant expression of beta-catenin was shown in normal cervical epithelium, while the aberrant expression rates of beta-catenin in CINI, CINII, CINIII and cervical carcinoma group were 28.6%, 40.0%, 62.5% and 84.2%. The aberrant expression rate of beta-catenin increased with the increase in degree of CIN and development of cervical carcinoma. The over-expression rate of EP-CAM was reversely related to the differentiation of cervical squamous cell carcinoma (P < 0.001). CONCLUSION: EP-CAM and beta-catenin may be involved in the carcinogenesis of squamous cell carcinoma of uterine cervix. The over-expression of EP-CAM and aberrant expression of beta-catenin may serve as markers of squamous carcinogenesis of uterine cervix.

[HPV detection and FHIT expression in esophageal squamous carcinoma from high incidence area in Cixian County].

BACKGROUND & OBJECTIVE: Cixian County is one of the high incidence areas of esophageal carcinoma in China. Up to now, no work has been conducted on the possible etiological role of human papilloma virus (HPV) infection on esophageal carcinoma in this local area. The aim of this study was to explore the putative roles of HPV infection on the esophageal carcinogenesis of the patients in Cixian and to elucidate the possible relationship between HPV existence and fragile histidine triad gene (FHIT gene, a putative tumor suppressor gene), expression in squamous cell carcinoma of esophagus. METHODS: The existence of HPV DNA and the expression of FHIT gene at protein level in esophageal carcinoma tissues were determined with PCR and immunohistochemical staining, respectively, in 128 archival paraffin-embedded tissue blocks of esophageal squamous carcinoma from the high incidence area of Cixian and 24 tissue blocks from the non-high incidence area. RESULTS: PCR results showed that the positive detection rate of HPV in esophageal carcinoma tissues from the high incidence area was 20.3%, which was slightly higher than that from the non-high incidence area (8.3%)(P >0.05). Immunohistochemically, aberrant expression of FHIT gene in esophageal carcinoma tissue was found in 75.6% cases from the high incidence area and only 54.2% from the non-high incidence area, which was significantly lower than that of the former (P< 0.05). No correlation was found between the abnormal expression of FHIT and existence of HPV DNA in esophageal squamous carcinoma tissues. CONCLUSION: HPV DNA could be detected in partial esophageal carcinoma cases from the high incidence area of esophageal carcinoma in Cixian. The aberrant expression rate of FHIT protein in the cases from the high incidence area of esophageal carcinoma in Cixian is higher than that from the non-high incidence area.