Kate Rittenhouse-Olson

Cancer vaccines and carbohydrate epitopes.

Tumor-associated carbohydrate antigens (TACA) result from the aberrant glycosylation that is seen with transformation to a tumor cell. The carbohydrate antigens that have been found to be tumor-associated include the mucin related Tn, Sialyl Tn, and Thomsen-Friedenreich antigens, the blood group Lewis related Lewis(Y), Sialyl Lewis(X) and Sialyl Lewis(A), and Lewis(X) (also known as stage-specific embryonic antigen-1, SSEA-1), the glycosphingolipids Globo H and stage-specific embryonic antigen-3 (SSEA-3), the sialic acid containing glycosphingolipids, the gangliosides GD2, GD3, GM2, fucosyl GM1, and Neu5GcGM3, and polysialic acid. Recent developments have furthered our understanding of the T-independent type II response that is seen in response to carbohydrate antigens. The selection of a vaccine target antigen is based on not only the presence of the antigen in a variety of tumor tissues but also on the role this antigen plays in tumor growth and metastasis. These roles for TACAs are being elucidated. Newly acquired knowledge in understanding the T-independent immune response and in understanding the key roles that carbohydrates play in metastasis are being applied in attempts to develop an effective vaccine response to TACAs. The role of each of the above mentioned carbohydrate antigens in cancer growth and metastasis and vaccine attempts using these antigens will be described.

Assignment of Weight-Based Antibody Units for 13 Serotypes to a Human Antipneumococcal Standard Reference Serum, Lot 89-S(F)

ABSTRACT Weight-based immunoglobulin G (IgG), IgM, IgA, and total Ig antibody assignments were made to human antipneumococcal standard reference serum lot 89-S, also known as lot 89-SF, for Streptococcus pneumoniae capsular polysaccharide (PnPs) serotypes 2, 6A, 8, 9N, 10A, 11A, 12F, 15B, 19A, 17F, 20, 22F, and 33F, as well as for C-polysaccharide (C-Ps), extending the standards usefulness for pneumococcal vaccine evaluation beyond the original serotype 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19F, and 23F assignments (S. A. Quataert, C. S. Kirch, L. J. Quackenbush Wiedl, D. C. Phipps, S. Strohmeyer, C. O. Cimino, J. Skuse, and D. V. Madore, Clin. Diagn. Lab. Immunol. 2:590-597, 1995). The additional 14 assignments were determined using an equivalence of absorbance method with an anti-PnPs serotype 6B reference enzyme-linked immunosorbent assay (EIA). To assure accuracy, anti-PnPs EIA for serotype 14 antibodies, a previously assigned serotype, was performed concurrently. This method assures consistency of the new microgram-per-microliter assignments with previous antiserotype assignments to lot 89-S. The sum of the experimentally derived isotype assignments for anti-PnPs serotypes in lot 89-S agrees well with the separately determined total Ig assignment for each serotype. The lot 89-S assignments for serotypes 1, 5, 6B, 14, 18C, 19F, and 23F were used for pneumococcal conjugate vaccine clinical trial evaluation and to generate data in efficacy trials where serological correlates for protection have been proposed. The assignment of antibody concentrations to additional pneumococcal serotypes in this reference reagent facilitates the consistent and accurate comparison of serum antibody concentrations across clinical trials.

Effect of particulate matter air pollution on C-reactive protein: a review of epidemiologic studies

Abstract Inflammatory response is implicated as a biologic mechanism that links particulate matter (PM) air pollution with health effects. C-reactive protein (CRP), an important acutephase reactant with profound proinflammatory properties, is used clinically as an indicator of the presence and intensity of inflammation. In vitro and in vivo animal studies suggest that CRP levels increase in response to PM exposure, but there was no consistency in epidemiologic studies. Herein, a systematic review was conducted to examine the association between PM exposure and serum CRP levels in humans. Elevated CRP levels were consistently found among children, and CRP elevations were also observed among healthy adults, albeit requiring higher peak levels of PM exposure. PM-induced CRP responses were not consistently found in adults with chronic inflammatory conditions, perhaps because of the use of anti-inflammatory medications in this population. Of the eight examined randomized trials, only one trial with a longer intervention period supported the effect of PM exposure on CRP concentrations. To provide conclusive evidence, further epidemiologic studies are needed to better quantify the magnitude of CRP level changes in response to PM with well-defined study populations and better control of various confounding factors.

Glycodendrimers: versatile tools for nanotechnology

Combining nanotechnology with glycobiology has triggered an exponential growth of research activities in the design of novel functional bionanomaterials (glyconanotechnology). More specifically, recent synthetic advances towards the tailored and versatile design of glycosylated nanoparticles namely glyconanoparticles, considered as synthetic mimetics of natural glycoconjugates, paved the way toward diverse biomedical applications. The accessibility of a wide variety of these structured nanosystems, in terms of shapes, sizes, and organized around stable nanoparticles have readily contributed to their development and applications in nanomedicine. In this context, glycosylated gold-nanoparticles (GNPs), glycosylated quantum dots (QDs), fullerenes, single-wall natotubes (SWNTs), and self-assembled glycononanoparticles using amphiphilic glycopolymers or glycodendrimers have received considerable attention to afford powerful imaging, therapeutic, and biodiagnostic devices. This review will provide an overview of the most recent syntheses and applications of glycodendrimers in glycoscience that have permitted to deepen our understanding of multivalent carbohydrate-protein interactions. Together with synthetic breast cancer vaccines, inhibitors of bacterial adhesions to host tissues including sensitive detection devices, these novel bionanomaterials are finding extensive relevance.

Computational Screening of the Human TF-Glycome Provides a Structural Definition for the Specificity of Anti-Tumor Antibody JAA-F11.

Recombinant antibodies are of profound clinical significance; yet, anti-carbohydrate antibodies are prone to undesirable cross-reactivity with structurally related-glycans. Here we introduce a new technology called Computational Carbohydrate Grafting (CCG), which enables a virtual library of glycans to be assessed for protein binding specificity, and employ it to define the scope and structural origin of the binding specificity of antibody JAA-F11 for glycans containing the Thomsen-Friedenreich (TF) human tumor antigen. A virtual library of the entire human glycome (GLibrary-3D) was constructed, from which 1,182 TF-containing human glycans were identified and assessed for their ability to fit into the antibody combining site. The glycans were categorized into putative binders, or non-binders, on the basis of steric clashes with the antibody surface. The analysis employed a structure of the immune complex, generated by docking the TF-disaccharide (Galβ1-3GalNAcα) into a crystal structure of the JAA-F11 antigen binding fragment, which was shown to be consistent with saturation transfer difference (STD) NMR data. The specificities predicted by CCG were fully consistent with data from experimental glycan array screening, and confirmed that the antibody is selective for the TF-antigen and certain extended core-2 type mucins. Additionally, the CCG analysis identified a limited number of related putative binding motifs, and provided a structural basis for interpreting the specificity. CCG can be utilized to facilitate clinical applications through the determination of the three-dimensional interaction of glycans with proteins, thus augmenting drug and vaccine development techniques that seek to optimize the specificity and affinity of neutralizing proteins, which target glycans associated with diseases including cancer and HIV.

Development and Characterization of Antibodies to Carbohydrate Antigens

Antibodies to carbohydrate antigens are critical for the study of bacteria, tumors, blood groups, and cell-cell adhesion interactions; for the analysis of viral, hormone, and toxin receptors; and, finally, for analysis of the glycosylation of recombinant proteins. However, antibodies to carbohydrate structures are more difficult to develop because of the T-cell-independent response to carbohydrates. This can result in the production of low affinity and difficult to work with IgM antibodies to these molecules. Screening technologies that include IgM antibodies can cause selections of antibodies with low-affinity binding sites because of the net avidity enhancement. Unfortunately, the low-affinity binding site can also have a similar affinity for unwanted structures. Production of antibodies using cellular extracts can result in antibodies that react with multiple related structures, and therefore the resultant bioassays have sensitivity or specificity problems. Protein conjugates of saccharides for the production of polyclonal and monoclonal antibodies to carbohydrate structures can be used to solve these problems. For monoclonal antibody development to oligosaccharides, mapping with closely related saccharides allows the determination of the areas of the saccharide to which the antibody binds so that conclusions can be made concerning which saccharide structures will cross-react. Determination of the reactivity of the produced antibodies with related saccharide structures is essential prior to utilization.

Preclinical studies with JAA-F11 anti-Thomsen-Friedenreich monoclonal antibody for human breast cancer.

AIM The Thomsen-Friedenreich antigen (TF-Ag) is a disaccharide hidden on normal cells, but selectively exposed on the surface of breast, colon, prostate and bladder cancer cells. JAA-F11, a highly specific monoclonal antibody to TF-Ag, reduces metastasis and prolongs survival in a mouse model. In addition,(124)I-JAA-F11 localizes 4T1 tumors in mice. These studies continue translation of JAA-F11 to human breast cancer. MATERIALS & METHODS & RESULTS: Of the 41 human breast cancer cell lines tested, 78% were positive for reactivity with JAA-F11 by whole-cell enzyme immunoassay and positivity occurred unrelated to estrogen, progesterone or HER2 receptor status. JAA-F11 inhibited the growth rate of the human cancer cell lines tested. At 1 h, approximately 80% of JAA-F11 internalized in the three cell lines tested. (124)I-JAA-F11 specifically imaged human triple-negative tumors in mice by microPET. CONCLUSION The results highlight the potential that humanized JAA-F11 may have for immunotherapy and drug conjugate therapy in breast cancer patients.

Synthesis and antibody binding properties of glycodendrimers bearing the tumor related T-antigen

Breast cancer marker T-antigen (Gal(β1-3)αGalNAc) was prepared as an allyl glycoside that was transformed into an active ester and coupled to a series of poly(amidoamine) dendrimers showing strong binding to mouse monoclonal IgG antibodies and serving as useful coating antigens in microtiter plates.

Jaa-f11: extending the life of mice with breast cancer.

JAA-F11 antibody (Ab) is a monoclonal Ab that is specific for the Thomsen–Friedenreich antigen, Galβ1-3GalNAcα (TF-Ag). TF-Ag, discovered in the late 1920s, is a tumor-associated carbohydrate Ag of many clinically widespread carcinomas. In a mouse model, JAA-F11 Ab significantly extended median survival time of animals with metastatic 4T1 breast tumors and caused > 50% inhibition of lung metastasis. 124Iodine labeled JAA-F11 Ab in in vivo micro positron emission tomography showed tumor specificity in a mouse breast tumor model, with no preferential uptake by any other organ. Human cancer cell adhesion to vascular endothelium was also blocked by JAA-F11. Structural specificity of the Ab was shown with glycan array analysis and indicated that this Ab, unlike many other Abs to TF-Ag, will not bind to a related glycolipid on natural killer cells, kidney or spleen. Patients with higher levels of naturally occurring anti-TF-Ag Ab appear to have a better prognosis, indicating that passive transfer of JAA-F11 or active immunization, resulting in production of anti-TF-Ag Ab, would clinically be beneficial for the patient.

Carbohydrate moieties as vaccine candidates: targeting the sweet spot in the immune response.

Advances in the use of carbohydrates as vaccine candidates for the prevention of infectious and malignant diseases was the topic for a meeting in Rockville, MD, sponsored by the National Institute of Allergy and Infectious Diseases involving a diverse group of scientists. Participants included research scientists and clinicians from academia and industry, and representatives from the National Institutes of Health and US Food and Drug Administration. This workshop was the third in a series of meetings designed to address issues relating to the immune response to carbohydrate antigens and how this information is used in the development of vaccines. Participants also identified roadblocks, research opportunities and resource needs. The meeting was organized into sessions that focused on recent advances in the immune response to microbial and cancer carbohydrate antigens, glycomics, novel vaccine approaches, novel adjuvants and delivery systems.