Heat labile enterotoxin of E. coli: a potential adjuvant for transcutaneous cancer immunotherapy

Escherichia coli heat labile enterotoxin (LT) has been shown to penetrate intact skin and to activate adaptive immunity. A nontoxic mutant, nLT, and its B subunit (LTB), have been evaluated separately for their potential use as a tool for transcutaneous delivery of antigens for cancer immunotherapy. We have shown that FITC-labeled nLT is taken up by human dendritic cells (hDC) in vitro and in mouse skin, and induces maturation and activation of hDC in vitro. hDC matured with nLT enhanced nonspecific melanoma antigen uptake and presentation to autologous CD8+ T cells. In mouse in vivo studies, nLT or LTB were applied on the skin either mixed with recombinant gp100 or genetically fused with a multiepitope polypeptide (MEP). Fused LTB-MEP induced antibody production that was dependent on LTB cell binding. We conclude that LT derivatives may be useful for the transcutaneous delivery of tumor antigens for cancer immunotherapy.     

With a careful look: still no low-level confound to face pop-out

In this issue of Vision Research, VanRullen, R. (2006). On second glance: Still no high-level pop-out effect for faces. Vision Research, in press. challenges our earlier Vision Research paper, “At first sight: A high-level pop-out effect for faces” (Hershler, O., & Hochstein, S. (2005). At first sight: A high-level pop-out effect for faces. Vision Research, 45, 1707-1724). In that paper, we showed that faces pop-out from a great variety of heterogeneous distractors. This search must have been based on a holistic combination of facial features, since it could not have relied on any single low-level distinguishing feature—each of which was present in at least some of the distractors. VanRullen implies that the pop-out effect is not limited to faces, is not holistic, and is due to a low-level confound, namely that the “low-level” Fourier amplitude spectrum may differentiate between faces and other categories. We now show that he fails to substantiate all three claims. His first experiment replicates our own and shows once again that faces do indeed pop-out, while other objects, such as cars, do not. The claim regarding the non-holistic nature of face search is based on a failure to differentiate between holistic processing for face detection and for individual face identification. His central claim is that the Fourier amplitude spectrum is processed low-level and could be used for face pop-out. However, changing the amplitude spectrum may well affect high-level representations as well. For example, his demonstration uses hybrid images which are extremely fuzzy, rendering them difficult to identify. More importantly, this claim would lead to the conclusion that targets with a non-face phase spectrum and only a face amplitude spectrum would pop-out among distractors with different amplitude spectra. We demonstrate that this is, of course, not the case and that the Fourier amplitude is not the hoped for “low-level confound”. Until another such “hidden” low level feature is found, we must accept that face pop out depends on a high level mechanism.     

Multiple adaptive mechanisms to chronic liver disease revealed at early stages of liver carcinogenesis in the Mdr2-knockout mice

Molecular events preceding the development of hepatocellular carcinoma were studied in the Mdr2-knockout (Mdr2-KO) mice. These mice lack the liver-specific P-glycoprotein responsible for phosphatidylcholine transport across the canalicular membrane. Portal inflammation ensues at an early age followed by hepatocellular carcinoma development after the age of 1 year. Liver tissue samples of Mdr2-KO mice in the early and late precancerous stages of liver disease were subjected to histologic, biochemical, and gene expression profiling analysis. In an early stage, multiple protective mechanisms were found, including induction of many anti-inflammatory and antioxidant genes and increase of total antioxidant capacity of liver tissue. Despite stimulation of hepatocyte DNA replication, their mitotic activity was blocked at this stage. In the late stage of the disease, although the total antioxidant capacity of liver tissue of Mdr2-KO mice was normal, and inflammation was less prominent, many protective genes remained overexpressed. Increased mitotic activity of hepatocytes resulted in multiple dysplastic nodules, some of them being steatotic. Expression of many genes regulating lipid and phospholipid metabolism was distorted, including up-regulation of choline kinase A, a known oncogene. Many other oncogenes, including cyclin D1, Jun, and some Ras homologues, were up-regulated in Mdr2-KO mice at both stages of liver disease. However, we found no increase of Ras activation. Our data suggest that some of the adaptive mechanisms induced in the early stages of hepatic disease, which protect the liver from injury, could have an effect in hepatocarcinogenesis at later stages of the disease in this hepatocellular carcinoma model.     

MUC1/X protein immunization enhances cDNA immunization in generating anti-MUC1 alpha/beta junction antibodies that target malignant cells

MUC1 has generated considerable interest as a tumor marker and potential target for tumor killing. To date, most antibodies against MUC1 recognize epitopes within the highly immunogenic alpha chain tandem repeat array. A major shortcoming of such antibodies is that the MUC1 alpha chain is shed into the peripheral circulation, sequesters circulating antitandem repeat array antibodies, and limits their ability to even reach targeted MUC1-expressing cells. Antibodies recognizing MUC1 epitopes tethered to the cell surface would likely be more effective. MUC1 alpha subunit binding the membrane-tethered beta subunit provides such an epitope. By use of a novel protocol entailing immunization with cDNA encoding full-length MUC1 (MUC1/TM) followed by boosting with the alternatively spliced MUC1/X isoform from which the tandem repeat array has been deleted, we generated monoclonal antibodies, designated DMC209, which specifically bind the MUC1 alpha/beta junction. DMC209 is exquisitely unique for this site; amino acid mutations, which abrogate MUC1 cleavage, also abrogate DMC209 binding. Additionally, DMC209 specifically binds the MUC1 alpha/beta junction on full-length MUC1/TM expressed by breast and ovarian cancer cell lines and on freshly obtained, unmanipulated MUC1-positive malignant plasma cells of multiple myeloma. DMC209 is likely to have clinical application by targeting MUC1-expressing cells directly and as an immunotoxin conjugate. Moreover, the novel immunization procedure used in generating DMC209 can be used to generate additional anti-MUC1 alpha/beta junction antibodies, which may, analogously to Herceptin, have cytotoxic activity. Lastly, sequential immunization with MUC1/TM cDNA acting as a nonspecific adjuvant followed by protein of interest may prove to be a generalizable method to yield high-titer specific antibodies.     

Heparanase promotes growth, angiogenesis and survival of primary breast tumors

Despite great strides toward diagnosis and therapy, breast cancer remains a most threatening disease in its incidence, morbidity and mortality; therefore, additional knowledge regarding the molecular mechanisms contributing to breast cancer progression, as well as new targets for drug discovery are highly needed. Heparanase is the predominant enzyme involved in cleavage of heparan sulfate, the main polysaccharide component of the extracellular matrix. Experimental and clinical data indicate that heparanase plays important roles in cancer metastasis and angiogenesis. In breast carcinoma patients, heparanase expression correlates with the metastatic potential of the tumor. The present study was undertaken to investigate the role of heparanase in local growth and angiogenesis of primary breast tumors. MCF-7 breast carcinoma cells were stable transfected with the human heparanase (H-hpa) cDNA, or empty vector (mock), and injected into the mammary pad of nude mice. MRI was applied to monitor progression of tumor growth and angiogenesis. We demonstrate that tumors produced by cells overexpressing heparanase grew faster and were 7-fold larger than tumors produced by mock transfected cells. This enhanced growth was accompanied by increased tumor vascularization and a higher degree of vessel maturation. Histological examination ascribed the differences in tumor growth to heparanase-stimulated cell proliferation and survival. In-vitro experiments reinforced heparanase role as a survival factor under stress conditions. Moreover, H-hpa tumor cells infiltrate into the adjacent stroma, promoting formation of highly vascularized fibrous bands. Our results emphasize the significance and clarify the involvement of heparanase in primary breast cancer progression by generating a supportive microenvironment that promotes tumor growth, angiogenesis and survival.     

Achalasia after vertical banded gastroplasty for morbid obesity: A case report

Achalasia is an unusual motility disorder that can be seen in conjunction with obesity. The prevalence of achalasia is unknown in obese patients and when present, the clinical characteristics are atypical. We report a case of achalasia that was diagnosed 13 years after a vertical-banded gastroplasty was performed.     

The effectiveness of Internet-based blood glucose monitoring system on improving diabetes control in adolescents with type 1 diabetes

Landau Z, Mazor‐Aronovitch K, Boaz M, Blaychfeld‐Magnazi M, Graph‐Barel C, Levek‐Motola N, Pinhas‐Hamiel O. The effectiveness of Internet‐based blood glucose monitoring system on improving diabetes control in adolescents with type 1 diabetes. Objective: To determine whether the use of an Internet‐based blood glucose monitoring system could improve glycemic control in adolescents with type 1 diabetes mellitus (T1DM). Methods: In a randomized, controlled clinical trial, a total of 70 adolescent subjects with T1DM were recruited. Subjects randomized to the intervention group (n = 36) were instructed to submit their blood glucose levels weekly by Internet to the Diabetes Care Team during a period of 6 months. Subjects randomized to the control group (n = 34) did not submit results but were under routine follow‐up. Results: At baseline, patients were 15.1 ± 2.6 years of age with mean HbA1c of 8.3 ± 1.3%. At the 6‐month follow‐up period, no by‐group differences in change from baseline to end of treatment HbA1c levels were detected. In the intervention group, 12/36 did not submit blood glucose levels and were classified as non‐compliant. In a secondary exploratory analysis in which non‐compliant patients were omitted, HbA1c values in the compliant intervention group declined from 8.5 ± 1.7% at baseline to 8.2 ± 1.2% at 6 months, while in the control group HbA1c values increased from 8.2 ± 1.1 to 8.4 ± 1.1%, this difference did not reach statistical significance. Conclusions: An Internet‐based blood glucose monitoring system was not associated with improved glycemic control in adolescents with T1DM. Identification of a sub‐group of compliant subjects who may improve metabolic control by using this tool is needed.     

Targeting the tumor microenvironment by immunotherapy: part 2

Cancer therapy was traditionally centered on the neoplastic cells. This included mainly surgery, radiation, and chemotherapy, in some cases hormone therapy and to a lesser extent immunotherapy--all traditionally targeted to the highly proliferating mutated tumor cells. In view of our present understanding of the powerfull influence of the tumor microenvironment (TME) on cancer behavior and response--and lack of response--to treatment, this previously ignored constituent of cancer now has to be considered as an important, even indispensable target for therapy. The TME may be targeted both to its immune and to its nonimmune components. The various immune evasion elements of the TME should be targeted as well.