Development of a Versatile Cancer Vaccine Format Targeting Antigen-Presenting Cells Using Proximity-Based Sortase A-Mediated Ligation of T-Cell Epitopes.

Cancer vaccines are a promising strategy to increase tumor-specific immune responses in patients who do not adequately respond to checkpoint inhibitors. Cancer vaccines that contain patient-specific tumor antigens are most effective but also necessitate the production of patient-specific vaccines. This study aims to develop a versatile cancer vaccine format in which patient-specific tumor antigens can be site-specifically conjugated by a proximity-based Sortase A (SrtA)-mediated ligation (PBSL) approach to antibodies that specifically bind to antigen-presenting cells to stimulate immune responses.

PD-L1 checkpoint blockade promotes regulatory T cell activity that underlies therapy resistance.

Despite the clinical success of immune checkpoint blockade (ICB), in certain cancer types, most patients with cancer do not respond well. Furthermore, in patients for whom ICB is initially successful, this is often short-lived because of the development of resistance to ICB. The mechanisms underlying primary or secondary ICB resistance are incompletely understood.

CRISPR/Cas9-based Engineering of Immunoglobulin Loci in Hybridoma Cells.

Development of the hybridoma technology by Köhler and Milstein (1975) has revolutionized the immunological field by enabling routine use of monoclonal antibodies (mAbs) in research and development efforts, resulting in their successful application in the clinic today. While recombinant good manufacturing practices production technologies are required to produce clinical grade mAbs, academic laboratories and biotechnology companies still rely on the original hybridoma lines to stably and effortlessly produce high antibody yields at a modest price. In our own work, we were confronted with a major issue when using hybridoma-derived mAbs: there was no control over the antibody format that was produced, a flexibility that recombinant production does allow.

Regulatory T Cell Depletion Using a CRISPR Fc-Optimized CD25 Antibody.

Regulatory T cells (T) are major drivers behind immunosuppressive mechanisms and present a major hurdle for cancer therapy. T are characterized by a high expression of CD25, which is a potentially valuable target for T depletion to alleviate immune suppression. The preclinical anti-CD25 (αCD25) antibody, clone PC-61, has met with modest anti-tumor activity due to its capacity to clear T from the circulation and lymph nodes, but not those that reside in the tumor.

Multiscale imaging of therapeutic anti-PD-L1 antibody localization using molecularly defined imaging agents.

Background: While immune checkpoint inhibitors such as anti-PD-L1 antibodies have revolutionized cancer treatment, only subgroups of patients show durable responses. Insight in the relation between clinical response, PD-L1 expression and intratumoral localization of PD-L1 therapeutics could improve patient stratification. Therefore, we present the modular synthesis of multimodal antibody-based imaging tools for multiscale imaging of PD-L1 to study intratumoral distribution of PD-L1 therapeutics.

Dual Site-Specific Chemoenzymatic Antibody Fragment Conjugation Using CRISPR-Based Hybridoma Engineering.

Functionalized antibodies and antibody fragments have found applications in the fields of biomedical imaging, theranostics, and antibody-drug conjugates (ADC). In addition, therapeutic and theranostic approaches benefit from the possibility to deliver more than one type of cargo to target cells, further challenging stochastic labeling strategies. Thus, bioconjugation methods to reproducibly obtain defined homogeneous conjugates bearing multiple different cargo molecules, without compromising target affinity, are in demand.

Functional diversification of hybridoma-produced antibodies by CRISPR/HDR genomic engineering.

Hybridoma technology is instrumental for the development of novel antibody therapeutics and diagnostics. Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted.

ICAM3-Fc Outperforms Receptor-Specific Antibodies Targeted Nanoparticles to Dendritic Cells for Cross-Presentation.

Optimal targeting of nanoparticles (NP) to dendritic cells (DCs) receptors to deliver cancer-specific antigens is key to the efficient induction of anti-tumour immune responses. Poly (lactic-co-glycolic acid) (PLGA) nanoparticles containing tètanus toxoid and gp100 melanoma-associated antigen, toll-like receptor adjuvants were targeted to the DC-SIGN receptor in DCs by specific humanized antibodies or by ICAM3-Fc fusion proteins, which acts as the natural ligand. Despite higher binding and uptake efficacy of anti-DC-SIGN antibody-targeted NP vaccines than ICAM3-Fc ligand, no difference were observed in DC activation markers CD80, CD83, CD86 and CCR7 induced.

Transient Peripapillary Retinoschisis in Glaucomatous Eyes.

. To investigate transient focal microcystic retinoschisis in glaucomatous eyes in images obtained with several imaging techniques used in daily glaucoma care. .

Genome-Wide Association Mapping of Fertility Reduction upon Heat Stress Reveals Developmental Stage-Specific QTLs in Arabidopsis thaliana.

For crops that are grown for their fruits or seeds, elevated temperatures that occur during flowering and seed or fruit set have a stronger effect on yield than high temperatures during the vegetative stage. Even short-term exposure to heat can have a large impact on yield. In this study, we used Arabidopsis thaliana to study the effect of short-term heat exposure on flower and seed development.

Accuracy of matching optic discs with visual fields: the European Structure and Function Assessment Trial (ESAFAT).

Purpose: To determine the ability of ophthalmologists across Europe to match stereoscopic optic disc photographs to visual fields of varying severity.

Design: Evaluation and comparison of 2 diagnostic tests.

Participants: A total of 109 of 260 invited ophthalmologists in 11 European countries.

RPE-normalized RNFL attenuation coefficient maps derived from volumetric OCT imaging for glaucoma assessment.

Purpose: We present spatial retinal nerve fiber layer (RNFL) attenuation coefficient maps for healthy and glaucomatous eyes based on optical coherence tomography (OCT) measurements. Quantitative analyses of differences between healthy and glaucomatous eyes were performed.

Methods: Peripapillary volumetric images of 10 healthy and 8 glaucomatous eyes were acquired by a Spectralis OCT system.

Surveillance for ocular hypertension: an evidence synthesis and economic evaluation.

Objectives: To determine effective and efficient monitoring criteria for ocular hypertension [raised intraocular pressure (IOP)] through (i) identification and validation of glaucoma risk prediction models; and (ii) development of models to determine optimal surveillance pathways.

Design: A discrete event simulation economic modelling evaluation. Data from systematic reviews of risk prediction models and agreement between tonometers, secondary analyses of existing datasets (to validate identified risk models and determine optimal monitoring criteria) and public preferences were used to structure and populate the economic model.

The mode of inheritance in tetraploid cut roses.

Tetraploid hybrid tea roses (Rosa hybrida) represent most of the commercial cultivars of cut roses and form the basis for breeding programmes. Due to intensive interspecific hybridizations, modern cut roses are complex tetraploids for which the mode of inheritance is not exactly known. The segregation patterns of molecular markers in a tetraploid mapping population of 184 genotypes, an F(1) progeny from a cross of two heterozygous parents, were investigated for disomic and tetrasomic inheritance.

The effect of glaucoma on the optical attenuation coefficient of the retinal nerve fiber layer in spectral domain optical coherence tomography images.

Purpose: To demonstrate the effect of glaucoma on the optical attenuation coefficient of the retinal nerve fiber layer (RNFL) in Spectral Domain Optical Coherence Tomography (SD-OCT) images.

Methods: We analyzed images of the peripapillary areas in 10 healthy and 30 glaucomatous eyes (mild, moderate, and advanced glaucoma, 10 eyes each), scanned with the Spectralis OCT (Heidelberg Engineering GmbH, Dossenheim, Germany). To calculate the RNFL attenuation coefficient (μ(att)), determined by the scattering properties of the RNFL, we used a model that normalized the reflectivity of the RNFL by the retinal pigment epithelium.

Automated segmentation by pixel classification of retinal layers in ophthalmic OCT images.

Current OCT devices provide three-dimensional (3D) in-vivo images of the human retina. The resulting very large data sets are difficult to manually assess. Automated segmentation is required to automatically process the data and produce images that are clinically useful and easy to interpret.

Early perfusion of a free RPE-choroid graft in patients with exudative macular degeneration can be imaged with spectral domain-OCT.

Purpose: To study early flow and revascularization in a free, autologous, retinal pigment epithelium (RPE)-choroid graft.

Methods: This prospective cohort study used spectral domain-optical coherence tomography (SD-OCT) after RPE-choroid graft surgery in 12 patients. This SD-OCT was combined with fluorescein angiography (FA) and indocyanine green angiography (ICGA) in 5 patients.

The ability of short-wavelength automated perimetry to predict conversion to glaucoma.

Purpose: Short-wavelength automated perimetry (SWAP) has been claimed to predict conversion to glaucoma 3 to 4 years before standard automated perimetry (SAP) defects occur. This study compared the moment of glaucomatous conversion between SWAP and SAP.

Design: Prospective, longitudinal follow-up study.

DNA barcoding discriminates the noxious invasive plant species, floating pennywort (Hydrocotyle ranunculoides L.f.), from non-invasive relatives.

Floating pennywort (Hydrocotyle ranunculoides L.f.), a member of the plant family Araliaceae originating from North America, is an example of an invasive aquatic species posing serious problems to the management of waterways outside of its original distribution area in Australia and Western Europe.

The prognosis of patients operated on for gastric cancer in relation to the percentage of lymph nodes invaded by tumour.

Introduction: In evaluating the type of gastrectomy and lymphadenectomy for gastric cancer, adequate prognosis has been dependant on the retrieval of at least 15 lymph nodes. We propose an alternative method in which the prognostic value is evaluated, according to whether or not more than 20% of the retrieved lymph nodes are invaded by tumour.

Materials & Methods: Sixty-five patients (36 men, 29 women) with a median age of 69 years (mean age 68.

Residual inorganic soil nitrogen in grass and maize on sandy soil.

Nitrogen (N) remaining as inorganic ('mineral') soil N at crop harvest (N(minH)) contributes to nitrate leaching. N(minH) data from 20 (grass) and 78 (maize) experiments were examined to identify main determinants of N(minH). N-rate (A) explained 51% (grass) and 34% (maize) of the variance in N(minH).

Alpha-gliadin genes from the A, B, and D genomes of wheat contain different sets of celiac disease epitopes.

Background: Bread wheat (Triticum aestivum) is an important staple food. However, wheat gluten proteins cause celiac disease (CD) in 0.5 to 1% of the general population.

Skin reactions due to low molecular weight heparin in pregnancy: a strategic dilemma.

Introduction: Heparin-induced allergic reactions may cause problems if heparin administration is needed for thrombo-embolic disease in pregnancy.

Case Reports: We report two cases of hypersensitivity to low molecular weight heparin (LMWH) in pregnancy.

Discussion: Alternative methods and new antithrombotic agents are discussed.

Ex-situ conservation of Black poplar in Europe: genetic diversity in nine gene bank collections and their value for nature development.

Populus nigra L. is a pioneer tree species of riparian ecosystems that is threatened with extinction because of the loss of its natural habitat. To evaluate the existing genetic diversity of P.