bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pleiotrophin Secreted from Human Breast Cancer MCF-7-Ptn Cells Activates Stromal Fibroblasts, Induces Epithelial Island Formation, and Remodels the Microenvironment.
Authors: Zuka Masahiko , Chang Yunchao , Wang Zhaoyi , Berenson James R. , Deuel Thomas F. 
Published: October 2019

Abstract
Pleiotrophin (PTN, Ptn) is an 18 kD cytokine that is expressed in many human breast cancers and its gene is inappropriately expressed in cell lines derived from these breast cancers. To study the siginificance of inappropriate expression of Ptn in human breast cancer cells on surrounding stromal cells, we first compared nude mouse xenografts of MCF-7 and MCF-7-Ptn cells. MCF-7-Ptn cells lack the Receptor Protein Tyrosine Phosphatase (RPTP)b/z, the PTN receptor, and thus are not responsive to PTN through autocrine or paracrine stimulation. The MCF-7-Ptn cell xenografts grew rapidly whereas MCF-7 cells xenografts were barely detectable 6 weeks after injection. MCF-7-Ptn cells that were co-injected with equal numbers of NIH3T3 cells grew even more rapidly in the flanks of the nude mice. Surprisingly, the MCF-7-Ptn cell explants developed a morphological phenotype remarkably similar to that of the human invasive ductal carcinoma. We then co-cultured MCF-7 cells that express Ptn (MCF-7-Ptn cells) with NIH 3T3 cells. Secretion of PTN from MCF-7-Ptn cells induced formation of sharply defined clusters of MCF-7-Ptn cells, termed “epithelial islands”, that were surrounded by dense fibrous bands interspersed with NIH 3T3 cells that morphologically closely resemble carcinoma associated fibroblasts (CAFs). A striking increase in tropoelastin and expression of type IV procollagen mRNA was identified in NIH3T3 cells co-cultured with MCF-7-Ptn cells. Furthermore, different markers often resulting from stromal cell-carcinoma cell interactions in breast cancer, including protein kinase C (PKC)-d, and both human and murine matrix metalloproteinase (MMP) 9 were identified either in cells or in the culture media taken from MCF-7-Ptn/NIH3T3 cell co-cultures. The induction of these biochemical and morphological features in the co-cultures of MCF-7-Ptn and NIH3T3 cells was demonstrated to be Ptn expression dependent, PTN-secretion dependent, and NIH3T3 cell dependent. The data suggest that PTN secretion alone from human breast cancer cells with inappropriate expression of Ptn is sufficient to markedly remodel the microenvironment of the breast cancer cell and induce a morphological transition of the MCF-7-Ptn cells and NIH3T3 cells to patterns resembling breast carcinomas through activation of the PTN/RPTPb/z signaling pathway in NIH3T3 cells and reciprocal signaling between the carcinoma stromal cells and the PTN secreting breast cancer cells.
Serum Adiponectin Compared with ZAP-70, CD38 and Immunoglobulin Heavy-Chain Mutation Status as a Predictor of Time to First Treatment in Early B-Cell Chronic Lymphocytic Leukemia.
Authors: Molica Stefano , Vitelli Gaetano , Cutrona Giovanna , Digiesi Giovanna , Mirabelli Rosanna , Giannarelli Diana , Sperduti Isabella , Ferrarini Manlio 
Published: October 2019

Abstract
We analyzed the correlation between well-established biological parameters of prognostic relevance in B-cell chronic lymphocytic leukemia [CLL] (i.e, mutational status of the immunoglobulin heavy chain variable region [IgVH], ZAP-70- and CD38-expression) and serum levels of adiponectin by evaluating the impact of these variables on the time to first treatment [TFT] in a series of 69 previously untreated Binet stage A B-cell CLL patients. Higher levels of adiponectin inversely correlated with peripheral blood lymphocytosis (r=−0.254; P=0.03), male gender (P=0.0002), percentage of ZAP−70-positive (r=− 0.285; P=0.04) and CD38-positive (r=−0.294; P=0.04) B-CLL cells.
After a median follow-up time of 32 months (range, 2–120 months) 28 (40.5%) out of 69 patients experienced a need for chemotherapy. Kaplan-Meier estimates of patients’ TFT, plotted after searching the best cut-off for adiponectin (i.e., 5.88 μg/mL ), demonstrated that low adiponectin concentration was associated with a shorter TFT (median TFT 32 months; P=0.01). The relationship among the various bio-pathological parameters, analyzed by the multiple correspondence analysis (MCA), showed two different clinico-biological profiles. The first, characterized by higher adiponectin serum levels (i.e., > 5.88 μg/mL), higher platelet count (> 174 x 109/L), lower β2-microglobulin [β2-m] (< 2.35 mg/L), presence of mutation in the IgVH and low percentage of either CD38-positive (< 20%) or ZAP-70-positive (< 20%) B-CLL cells, was associated with a stable pattern of disease generally not requiring therapy. The second, defined by lower adiponectin levels, lower platelet count, high β2-m concentration, absence of mutation in the IgVH, high percentage of CD38- and ZAP-70 positive B-cells, was associated with a more progressive pattern of disease and a shorter TFT. The univariate Cox proportional hazard model demonstrated that in addition with lower serum levels of adiponectin (Hazard Ratio [HR], 2.936; P=0.01), the absence of IgVH mutational status (HR, 6.378; P=0.002) and ZAP-70-positivity (HR, 3.314; P=0.02) were associated with a shorter TFT. However, in multivariate analysis only ZAP-70 positivity emerged as predictor of the TFT (HR, 5.187; P=0.008).
Our results indicate that in early B-cell CLL clinico-biological profile including among other parameters adiponectin may provide a useful insight into the complex interrelationship of prognostic variables and semplify their interpretation. However, adiponectin may not replace the need for the determination of ZAP-70 and IgVH mutational status.
Vaccine Properties of a Novel Marker Gene-Free Recombinant Modified Vaccinia Ankara (MVA) Expressing Immunodominant CMV Antigens.
Authors: Diamond Don J. , Wang Zhongde , La Rosa Corinna , Srivastava Tumul , Zhou Wendy , Li Zhongqi , Ly Heang , Akbarnejad Tamineh , Krishnan Aparna , Britt William J. 
Published: October 2019

Abstract
Human CMV continues to be a major risk factor in patients undergoing allogeneic hematopoietic stem cell (HSCT) and solid organ transplant (SOT), despite advancement of antiviral therapy. CMV tegument protein pp65 and CMV immediate early gene product IE1 are both considered immunodominant targets of cell-mediated immunity (CMI) and potentially capable of controlling CMV infection. Similar to healthy adults, CMV tegument protein pp65 and IE1 are frequently recognized by human CD4 and CD8 T cells in HCT and SOT recipients. To better assess their role in host defense, we have constructed a novel attenuated poxvirus (MVA) transfer vector named pZWIIA and generated a recombinant MVA (rMVA) expressing both full-length pp65 and exon4 of IE1 (pp65-IE1-MVA) at high levels, followed by the genetic removal of the bacterial marker gene used to distinguish recombinant forms during the screening process. Immunogenicity evaluation indicates that pp65-IE1-MVA not only can induce robust primary CMI to both antigens in HLA A2.1, B7 and A11 transgenic mice, but also can stimulate vigorous expansion of memory T lymphocyte responses to pp65 and IE1 in PBMC of CMV-positive donors. The recent discovery that additional early antigens of CMV are also well-recognized by healthy adults prompted us to include the IE2 antigen in the vaccine. Evaluation of in vivo immunogenicity of the vaccine expressing IE2 in both transgenic mice and in vitro antigenicity in human peripheral blood lymphocytes of HSCT and SOT recipients will be presented. A novel overlapping peptide library spanning full-length IE2 has been constructed in our laboratory, and used in conjunction with the vaccine expressing IE2, along with IE1 and pp65. Since MVA has a foreign gene capacity of over 30 kilobases (Kb), we have developed a strategy to maximize coverage of haplotypes through insertion of 6 additional antigens into novel insertion sites of the virus, for a total of just 12.0 Kb of the CMV genome. The selected antigens cover 83% of the population, while representing over 22% of the combined CD4 and CD8 T cell repertoire in seropositives. Our studies of the recognition of these antigens expressed in MVA and monitored with our own synthesized over-lapping peptide libraries in both HSCT and SOT recipients will be presented. These properties make the MVA-based vaccine ideal for the dual role of priming and boosting CMV-specific T cell immunity as a means to control CMV disease in recipients of HCT or SOT. pZWIIA alone or in combination with other MVA transfer vectors can be used to generate MVA based multiple-antigen vaccine which have application in vaccine development for a wide spectrum of infectious diseases and cancer.
Early Immunoglobulin Free Light Chain (FLC) Response Post Autologous Peripheral Blood Stem Cell Transplant Predicts for Hematologic Complete Response in Patients with Multiple Myeloma.
Authors: Dispenzieri Angela , Rajkumar S. Vincent , Plevak Matthew F. , Katzmann Jerry A. , Kyle Robert A. , Larson Dirk , Lacy Martha Q. , Clark Raynell , DeGoey Roberta , Hayman Suzanne R. , Kumar Shaji K. , Gertz Morie Abraham 
Published: October 2019

Abstract
Background: FLC is a valuable diagnostic tool to monitor response in patients with AL and oligosecretory MM. It is also prognostic in patients with MGUS. Its value in patients with non-oligosecretory myeloma has not been defined. Because of the short half-life of free-light chains, an early drop in free-light chain might be predictive for long term outcome post PBSCT.
Methods: Between 8/1/03 and 12/10/04, 45 patients with multiple myeloma and abnormal FLC ratios (less than 02.6 or greater 1.65), who were undergoing PBSCT consented to have waste serum from routine post-transplant daily blood draws collected and frozen for future FLC analysis. The immunoglobulin FLC assay (Binding Site, U.K.) was run on samples collected from day +1 until patients were dismissed from our transplant center. Hematologic response was coded according to standard IBMTR criteria at approximately 3 months status post transplant (BJH 1998). Two types of immunoglobulin FLC response were coded: 50% reduction (FLC50) and 90% reduction (FLC90) of the difference of the involved and uninvolved FLC (Leukemia 2006). Two time periods of FLC response were analyzed: early (day 7); and delayed (day 19, range 13 to 24). Endpoints analyzed were overall response (>=PR), complete response rate (CR), progression free survival (PFS), and overall survival (OS).
Results: The median age of the myeloma group was 59 years, and 53% were male. Thirty-three patients were monoclonal kappa, and 12 lambda. Forty-two percent were transplanted in response/plateau, 22% were primary refractory, and 36% relapsed disease. The overall hematologic response rate was 93% with a CR rate of 33%. Twenty-five patients have relapsed at a median of 14.2 months. Twelve patients have died, and median overall survival for the group has not been reached at a median follow-up of 24.6 months. Forty-nine and 58 percent of patients had early and delayed FLC90, respectively. Early FLC90, but not delayed FLC90, predicted for complete response (p<0.001). Neither early nor delayed FLC90 predicted for either PFS or OS. In addition, 89% and 96% of patients had an early and delayed FLC50, respectively. FLC50 at either time point did not predict for overall response, CR, PFS, or OS.
Conclusion: Immunoglobulin free light chain responses are seen within the first weeks post high dose chemotherapy with peripheral blood stem cell transplant. A 90% reduction in FLC seen within the first seven days predicts for complete response. However, this does not yet translate into prolonged PFS or OS, but with longer follow-up separation of the survival curves may be seen.
Toll-Like Receptors 2 and 4 Are Involved in the Induction of Graft-Versus-Host-Disease in Mice.
Authors: Nogai Axel , Heimesaat Markus M. , Thiele Marc , Bereswill Stefan , Thiel Eckhard , Goebel Ulf B. , Uharek Lutz 
Published: October 2019

Abstract
BACKGROUND: Intestinal Graft-versus-Host disease is a frequent and often lethal complication after allogenic stem cell transplantation. Since NOD2 polymorphisms have been recognized as potential triggers of severe intestinal GvHD in humans, we have developed murine transplantation models to investigate the role of different pattern recognition receptors (PRR) in GvHD and GvL. Here we report our results on the role of TLR2 and TLR4 for the induction of GvHD.
METHODS: Severity of GvHD in wildtype (wt) C57B/10 (H-2Db), TLR2−/−, TLR4−/−, and combined TLR2−/−TLR4−/− C57B/10 mice was investigated. Mice received treosulfan 2000 mg/kg from day -3 to -1 and cyclophosphamide 200 mg/kg day -1 prior to injection of 10×10^6 H-2Dd BM cells and 5×10^6 splenocytes (SC). Survival and GvHD score were assessed daily. Engraftment was determined every 2 weeks in pB and at the end of the experiments in bone marrow by flow cytometry. T cell alloreactivity in GvH direction was assessed by MLR using splenocytes as stimulators from PRR-deficient mice or wt as control and CFSE-staining as read-out. The relevance of PRR ligands for the enhancement of GvH alloreactivity was determined by addition of lipid A, lipopetides, or CpG.
RESULTS:
in vivo data: The transfer of 10×10^6 BMC + 5×10^6 SC induced a severe GvHD in all wt recipients, leading to death of 90% of the animals within 20 days. Recipient mice lacking either TLR2 or TLR4 showed only a slightly and not significantly decreased GvHD lethality. In recipients lacking both PPRs, i.e. TLR2 and TLR4, GvHD was generally milder and the majority (60%) of the animals survived until day 20 (p<0.05). However, the long term survival was not significantly improved. Differences in clinical severity of GvHD were confirmed histologically. In vitro data: Stimulation with cells from TLR2−/− and TLR4−/− mice resulted in a decreased alloreactivity in MLR. A median of 2% of Balb/c CD4+ T cells proliferated in response to C57B/10 stimulators. The addition of the TLR2 and TLR4 ligands lipopeptide, Lipid A and CpG significantly (p<0.05) increased the proliferation of CD4+ T cells in a specific manner more than twofold.
CONCLUSION: Our in vivo and in vitro data consistantly show that bacterial components are involved in triggering GvH alloreactivity via different types of PPRs. Binding of bacterial substances to TLR2 and TLR4 leads to activation of the immune system and subsequent induction of GvHD. Our data provide an experimental basis for the development of strategies for modulation of the intestinal gut flora by selective gut decontamination and/or probiotic regimens to prevent GvHD in humans.
Mechanism of Alemtuzumab Depletion of T Cells in Stem Cell Products.
Authors: Gulen Dumrul , Bociek Robert G. , Coskun Ugur , Devetten Marcel P. , Talmadge James E. 
Published: October 2019

Abstract
Alemtuzumab (a humanized murine monoclonal antibody [MAb] against CD52) is an effective drug for in vitro T cell purging of allogeneic stem cell products when a product is incubated with 10mg of antibody for 30 minutes prior to infusion. Recovery of CD34+ cells has been excellent and T cell purging with alemtuzumab has resulted in a decreased incidence of grade 2–4 GVHD and a reduced incidence of extensive GVHD and a low overall risk of graft failure. However, a higher incidence of cytomegalovirus (CMV) reactivation, respiratory and polyoma virus infections have been observed as has an increased relapse rate in patients with chronic myelogenous leukemia. In vitro purging, using alemtuzumab and infusion of unwashed products, has been associated with a reduced incidence of clinically significant GVHD, an increase in infectious complications and a possible increase in relapse rates. The present studies optimize the purging protocol and examine the mechanism of alemtuzumab purging to identify the role of antibody dependant cell mediated cytotoxicity (ADCC) versus antibody dependant complement mediated cytotoxicity (Ab–C’). Both ADCC and Ab–C’ mediated cytotoxicity were active, although ADCC was noticeably more effective. However, cellular viability was improved by incubation with 10% autologous serum. A two hour incubation (1–2×01e8 cells/ml) resulted in a 73% reduction in CD3+ cells, which was increased to 85% after a 16 hour co-incubation. In addition, natural killer (NK) and B cells were purged with alemtuzumab; as were lineage negative (Lin−)-HLA-DR+CD123+ dendritic cells (DCs) but not Lin-HLA-DR+CD11c+ DCs. Immature myeloid suppressor cells (IMSCs), which are Lin−DR−CD33+ or DR−CD14−CD11b+, have a low sensitivity to alemtuzumab cytotoxicity as did hematopoietic progenitor cells (CFU-c function or CD34+ number). This is an exciting observation as IMSCs have the potential to induce T cell tolerance. Optimal T cell purging, with retention of CD34+ hematopoietic stem cell number and activity, requires a 4–6 hr co-incubation of 2×10e8/ml hematopoietic stem cells, 100 ug/ml alemtuzumab in 10% autologous serum at 4C and removal of alemtuzumab by washing in Plasma-Lyte. Co-incubation at 4C and the presence of 10% autologous serum, was required to minimize the toxicity of a 24 hr co-incubation using a high frequency (1–2×10e8/ml) of hematopoietic cells. The selective depletion of T cells, with a retention of IMSCs, suggests that such a purging protocol has potential to not only reduce acute and chronic GVHD but to also induce major histocompatibility complex tolerance. However, our clinical studies need to be completed in order to assess the impact on leukemic relapse.
Interest of FDG-PET Scanning for the Monitoring of Immunotherapies for Follicular Lymphoma.
Authors: Baudard Marion , Comte Frederic , Conge Anne Marie , Mariano-Goulart Denis , Klein Bernard , Rossi Jean-Francois 
Published: October 2019

Abstract
Except in the subset of patients with localized disease, follicular lymphoma (FL), the most common type of indolent non-Hodgkin’s lymphoma, should still be regarded as an incurable malignancy with a relentless relapsing course. New therapeutic approaches have been developed including anti-tumor vaccination strategies with induced humoral and cellular immune responses revealed by sensitive biological analyses but no imaging of the lymphocyte activation developed so far. Because [18F]fluorodeoxyglucose (FDG) is sequestred in metabolically active cells, we prospectively investigated the use of FDG-PET (positron emission tomography) to kinetically monitor cell activation in 7 FL patients vaccinated by repeated intradermal (ID) and intravenous injections of tumor lysate-pulsed autologous dendritic cells (DC-lysate). Patients were administered 5 vaccines and all patients developed a positive delayed-type hypersensitivity reaction at the site of DC-lysate ID injections after several vaccines administration, testifying of the induction of a biological response. Among the 4 patients with measurable disease at time of vaccination, 2 complete remissions (CR) and 2 disease progressions were observed. Three patients with initial normal computed tomographies (CT) were still in CR 12 months after the last vaccine. PETs were performed before vaccination, 14 days and 1 month after the last vaccine. PET revealed initial malignant nodes infracentimetric on concomitant CT, and was very sensitive in documenting disease progression even in patients with no significant variations of CT findings. A transient marked increase in FDG uptake within malignant nodes was observed early after vaccine administration in a patient who achieved a CR (fig 1-3-4-5), while ranges of tracer uptake variations were less important with a continuous increase of FGD uptake, size and number of involved loci in non responding patients (fig 2: follow up of intensity of FDG uptake by 6 tumoral nodes in a non responding pt). In conclusion, FDG-PET is a non invasive imaging procedure which besides being useful for the staging and the evaluation of treatment efficacy in FL, might be of crucial interest to detect cell responses induced by immunotherapies.
Figure Figure
Induction of Polyclonal T-Cell Responses Against Cutaneous T-Cell Lymphomas by Autologous Dendritic Cells Transfected with Amplified Tumor RNA.
Authors: Ni Xiao , Richmond Heather , Shpall Elizabeth , Duvic Madeleine 
Published: October 2019

Abstract
The indolent course of cutaneous T-cell lymphoma (CTCL) patients makes them attractive candidates for immunotherapeutic strategies. However, tumor specific antigens are unknown in CTCL, and thus this is an obstacle to the development of CTCL vaccines. Recently, vaccinations with the tumor antigen repertoire has shown to be a superior strategy to elicit protective immunity for hematological malignancies since it does not require definition of tumor antigens or HLA haplotyping (Borrello IM, Sotomayor EM. Cancer Control, 9:138–151,2002). Studies have shown that dendritic cells (DC) transfected with tumor mRNA amplified from primary tumor cells can stimulate anti-tumor T cell responses (Boczkowski et al, Cancer Research, 60: 1028–1034,2000; Muller et al, Blood, 103: 1763–1769, 2004). In this study, we determined the efficacy of ex vivo induction of anti-CTCL T cell responses using autologous DCs transfected with tumor RNA amplified from CTCL cells. We established a protocol for amplification of whole CTCL mRNA. DCs were generated from CD14+monocytes of patients with CTCL, transfected with amplified tumor RNA (5μg/106), matured with IL-1β, TNF-α, IL-6, and PGE2, and co-cultured with autologous peripheral blood lymphocytes. Induction of CTCL-specific cytotoxic T lymphocytes (CTL), determined by IFN-γ and Granzyme B secretion, was measured by ELISPOT assays. As a result, DCs transfected with amplified tumor RNA induced a 1–3 fold increase in specific IFN-γ secreting and Granzyme B releasing CTLs, compared to control DCs that were not transfected, in 3 of 5 patients. There was only an induction of granzyme B releasing CTLs seen in the fourth patient. There was no induction of either IFN-γ secreting or granzyme B releasing CTLs in the fifth CTCL patient who had low CD8+ T cells in the peripheral blood. These data suggest that vaccinations using DCs transfected with amplified tumor RNA might be a potent new strategy in the treatment of CTCL. It provides the rationale for further investigation of a potentially clinically effective and broadly applicable treatment for CTCL patients, and has the advantage of not requiring large amounts of tumor cells or characterization of a specific relevant antigen in each patient.