Generation of a Fully Human scFv that binds Tumor-Specific Glycoforms

Tumor-specific glycosylation changes are an attractive target for the development of diagnostic and therapeutic applications. Periostin is a glycoprotein with high expression in many tumors of epithelial origin including ovarian cancer. Strategies to target the peptide portion of periostin as a diagnostic or therapeutic biomarker for cancer are limited due to increased expression of periostin in non-cancerous inflammatory conditions. Here, we have screened for antibody fragments that recognize the tumor-specific glycosylation present on glycoforms of periostin containing bisecting N-glycans in ovarian cancer using a yeast-display library of antibody fragments, while subtracting those that bind to the periostin protein with glycoforms found in non-malignant cell types. We generated a biotinylated form of a fully human scFv antibody (scFvC9) that targets the bisecting N-glycans expressed by cancer cells. Validation studies in vitro and in vivo using scFvC9 indicate this antibody can be useful for the development of diagnostic, imaging, and therapeutic applications for cancers that express the antigen

Use of cancer-specific yeast-secreted in vivo biotinylated recombinant antibodies for serum biomarker discovery

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Trumpler 16-26: A New Centrifugal Magnetosphere Discovered via SDSS/APOGEE H-band Spectroscopy

We report the discovery of a new example of the rare class of highly magnetized, rapidly rotating, helium enhanced, early B stars that produce anomalously wide hydrogen emission due to a centrifugal magnetosphere (CM). The star is Trumpler 16-26, a B1.5 V member of the Trumpler 16 open cluster. A CM was initially suspected based on hydrogen Brackett series emission observed in SDSS/APOGEE $H$-band spectra. Similar to the other stars of this type, the emission was highly variable and at all times remarkable due to the extreme velocity separations of the double peaks (up to 1300 km s$^{-1}$.) Another clue lay in the TESS lightcurve, which shows two irregular eclipses per cycle when phased with the likely 0.9718115 day rotation period, similar to the behavior of the well known CM host star $\sigma$ Ori E. To confirm a strong magnetic field and rotation-phase-locked variability, we initiated a follow-up campaign consisting of optical spectropolarimetry and spectroscopy. The associated data revealed a longitudinal magnetic field varying between $-3.1$ and $+1.6$ kG with the period found from photometry. The optical spectra confirmed rapid rotation ($v \sin i=195$ km s$^{-1}$), surface helium enhancement, and wide, variable hydrogen emission. Tr16-26 is thus confirmed as the 20$^{\rm th}$ known, the fourth most rapidly rotating, and the faintest CM host star yet discovered. With a projected dipole magnetic field strength of $B_{\rm d}>11$ kG, Tr16-26 is also among the most magnetic CM stars

Safety, tumor trafficking and immunogenicity of chimeric antigen receptor (CAR)-T cells specific for TAG-72 in colorectal cancer.

BackgroundT cells engineered to express chimeric antigen receptors (CARs) have established efficacy in the treatment of B-cell malignancies, but their relevance in solid tumors remains undefined. Here we report results of the first human trials of CAR-T cells in the treatment of solid tumors performed in the 1990s.MethodsPatients with metastatic colorectal cancer (CRC) were treated in two phase 1 trials with first-generation retroviral transduced CAR-T cells targeting tumor-associated glycoprotein (TAG)-72 and including a CD3-zeta intracellular signaling domain (CART72 cells). In trial C-9701 and C-9702, CART72 cells were administered in escalating doses up to 1010 total cells; in trial C-9701 CART72 cells were administered by intravenous infusion. In trial C-9702, CART72 cells were administered via direct hepatic artery infusion in patients with colorectal liver metastases. In both trials, a brief course of interferon-alpha (IFN-α) was given with each CART72 infusion to upregulate expression of TAG-72.ResultsFourteen patients were enrolled in C-9701 and nine in C-9702. CART72 manufacturing success rate was 100% with an average transduction efficiency of 38%. Ten patients were treated in CC-9701 and 6 in CC-9702. Symptoms consistent with low-grade, cytokine release syndrome were observed in both trials without clear evidence of on target/off tumor toxicity. Detectable, but mostly short-term (≤14 weeks), persistence of CART72 cells was observed in blood; one patient had CART72 cells detectable at 48 weeks. Trafficking to tumor tissues was confirmed in a tumor biopsy from one of three patients. A subset of patients had 111Indium-labeled CART72 cells injected, and trafficking could be detected to liver, but T cells appeared largely excluded from large metastatic deposits. Tumor biomarkers carcinoembryonic antigen (CEA) and TAG-72 were measured in serum; there was a precipitous decline of TAG-72, but not CEA, in some patients due to induction of an interfering antibody to the TAG-72 binding domain of humanized CC49, reflecting an anti-CAR immune response. No radiologic tumor responses were observed.ConclusionThese findings demonstrate the relative safety of CART72 cells. The limited persistence supports the incorporation of co-stimulatory domains in the CAR design and the use of fully human CAR constructs to mitigate immunogenicity

B fields in OB stars (BOB): on the detection of weak magnetic fields in the two early B-type stars beta CMa and epsilon CMa

Within the context of the "B fields in OB stars (BOB)" collaboration, we used the HARPSpol spectropolarimeter to observe the early B-type stars beta CMa (HD44743; B1 II/III) and epsilon CMa (HD52089; B1.5 II). For both stars, we consistently detected the signature of a weak (<30 G in absolute value) longitudinal magnetic field. We determined the physical parameters of both stars and characterise their X-ray spectrum. For beta CMa, our mode identification analysis led to determining a rotation period of 13.6+/-1.2 days and of an inclination angle of the rotation axis of 57.6+/-1.7 degrees, with respect to the line of sight. On the basis of these measurements and assuming a dipolar field geometry, we derived a best fitting obliquity of ~22 degrees and a dipolar magnetic field strength (Bd) of ~100 G (60<Bd<230 G within 1 sigma), below what is typically found for other magnetic massive stars. For epsilon CMa we could only determine a lower limit on the dipolar magnetic field strength of 13 G. For this star, we determine that the rotation period ranges between 1.3 and 24 days. Both stars are expected to have a dynamical magnetosphere. We also conclude that both stars are most likely core hydrogen burning and that they have spent more than 2/3 of their main sequence lifetime. A histogram of the distribution of the dipolar magnetic field strength for the magnetic massive stars known to date does not show the magnetic field "desert" observed instead for intermediate-mass stars. The biases involved in the detection of (weak) magnetic fields in massive stars with the currently available instrumentation and techniques imply that weak fields might be more common than currently observed. Our results show that, if present, even relatively weak magnetic fields are detectable in massive stars and that more observational effort is probably still needed to properly access the magnetic field incidence.Comment: Accepted for publication on A&A. The astroph abstract has been shortened compared to that of the pdf fil

The Role of Innate APOBEC3G and Adaptive AID Immune Responses in HLA-HIV/SIV Immunized SHIV Infected Macaques

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A Mouse to Human Search for Plasma Proteome Changes Associated with Pancreatic Tumor Development

Background: The complexity and heterogeneity of the human plasma proteome have presented significant challenges in the identification of protein changes associated with tumor development. Refined genetically engineered mouse (GEM) models of human cancer have been shown to faithfully recapitulate the molecular, biological, and clinical features of human disease. Here, we sought to exploit the merits of a well-characterized GEM model of pancreatic cancer to determine whether proteomics technologies allow identification of protein changes associated with tumor development and whether such changes are relevant to human pancreatic cancer. Methods and Findings: Plasma was sampled from mice at early and advanced stages of tumor development and from matched controls. Using a proteomic approach based on extensive protein fractionation, we confidently identified 1,442 proteins that were distributed across seven orders of magnitude of abundance in plasma. Analysis of proteins chosen on the basis of increased levels in plasma from tumor-bearing mice and corroborating protein or RNA expression in tissue documented concordance in the blood from 30 newly diagnosed patients with pancreatic cancer relative to 30 control specimens. A panel of five proteins selected on the basis of their increased level at an early stage of tumor development in the mouse was tested in a blinded study in 26 humans from the CARET (Carotene and Retinol Efficacy Trial) cohort. The panel discriminated pancreatic cancer cases from matched controls in blood specimens obtained between 7 and 13 mo prior to the development of symptoms and clinical diagnosis of pancreatic cancer. Conclusions: Our findings indicate that GEM models of cancer, in combination with in-depth proteomic analysis, provide a useful strategy to identify candidate markers applicable to human cancer with potential utility for early detection

HE4 and CA125 as a diagnostic test in ovarian cancer: prospective validation of the Risk of Ovarian Malignancy Algorithm

BACKGROUND: Recently, a Risk of Ovarian Malignancy Algorithm (ROMA) utilising human epididymis secretory protein 4 (HE4) and CA125 successfully classified patients as presenting a high or low risk for epithelial ovarian cancer (EOC). We validated this algorithm in an independent prospective study. METHODS: Women with a pelvic mass, who were scheduled to have surgery, were enrolled in a prospective study. Preoperative serum levels of HE4 and CA125 were measured in 389 patients. The performance of each of the markers, as well as that of ROMA, was analysed. RESULTS: When all malignant tumours were included, ROMA (receiver operator characteristic (ROC)-area under curve (AUC) = 0.898) and HE4 (ROC-AUC) = 0.857) did not perform significantly better than CA125 alone (ROC-AUC = 0.877). Using a cutoff for ROMA of 12.5% for pre-menopausal patients, the test had a sensitivity of 67.5% and a specificity of 87.9%. With a cutoff of 14.4% for post-menopausal patients, the test had a sensitivity of 90.8% and a specificity of 66.3%. For EOC vs benign disease, the ROC-AUC of ROMA increased to 0.913 and for invasive EOC vs benign disease to 0.957. CONCLUSION: This independent validation study demonstrated similar performance indices to those recently published. However, in this study, HE4 and ROMA did not increase the detection of malignant disease compared with CA125 alone. Although the initial reports were promising, measurement of HE4 serum levels does not contribute to the diagnosis of ovarian cancer. British Journal of Cancer (2011) 104, 863-870. doi:10.1038/sj.bjc.6606092 www.bjcancer.com Published online 8 February 2011 (C) 2011 Cancer Research U