@article{10836, author = {Pranger, Christina L. and Fazekas-Singer, Judit and Köhler, Verena K. and Pali‐Schöll, Isabella and Fiocchi, Alessandro and Karagiannis, Sophia N. and Zenarruzabeitia, Olatz and Borrego, Francisco and Jensen‐Jarolim, Erika}, issn = {1398-9995}, journal = {Allergy}, keywords = {Immunology, Immunology and Allergy}, number = {5}, pages = {1553--1556}, publisher = {Wiley}, title = {{PIPE‐cloned human IgE and IgG4 antibodies: New tools for investigating cow's milk allergy and tolerance}}, doi = {10.1111/all.14604}, volume = {76}, year = {2021}, } @article{7864, abstract = {Purpose of review: Cancer is one of the leading causes of death and the incidence rates are constantly rising. The heterogeneity of tumors poses a big challenge for the treatment of the disease and natural antibodies additionally affect disease progression. The introduction of engineered mAbs for anticancer immunotherapies has substantially improved progression-free and overall survival of cancer patients, but little efforts have been made to exploit other antibody isotypes than IgG. Recent findings: In order to improve these therapies, ‘next-generation antibodies’ were engineered to enhance a specific feature of classical antibodies and form a group of highly effective and precise therapy compounds. Advanced antibody approaches include among others antibody-drug conjugates, glyco-engineered and Fc-engineered antibodies, antibody fragments, radioimmunotherapy compounds, bispecific antibodies and alternative (non-IgG) immunoglobulin classes, especially IgE. Summary: The current review describes solutions for the needs of next-generation antibody therapies through different approaches. Careful selection of the best-suited engineering methodology is a key factor in developing personalized, more specific and more efficient mAbs against cancer to improve the outcomes of cancer patients. We highlight here the large evidence of IgE exploiting a highly cytotoxic effector arm as potential next-generation anticancer immunotherapy.}, author = {Singer, Judit and Singer, Josef and Jensen-Jarolim, Erika}, issn = {14736322}, journal = {Current opinion in allergy and clinical immunology}, number = {3}, pages = {282--289}, publisher = {Wolters Kluwer}, title = {{Precision medicine in clinical oncology: the journey from IgG antibody to IgE}}, doi = {10.1097/ACI.0000000000000637}, volume = {20}, year = {2020}, } @article{8225, abstract = {Birch pollen allergy is among the most prevalent pollen allergies in Northern and Central Europe. This IgE-mediated disease can be treated with allergen immunotherapy (AIT), which typically gives rise to IgG antibodies inducing tolerance. Although the main mechanisms of allergen immunotherapy (AIT) are known, questions regarding possible Fc-mediated effects of IgG antibodies remain unanswered. This can mainly be attributed to the unavailability of appropriate tools, i.e., well-characterised recombinant antibodies (rAbs). We hereby aimed at providing human rAbs of several classes for mechanistic studies and as possible candidates for passive immunotherapy. We engineered IgE, IgG1, and IgG4 sharing the same variable region against the major birch pollen allergen Bet v 1 using Polymerase Incomplete Primer Extension (PIPE) cloning. We tested IgE functionality and IgG blocking capabilities using appropriate model cell lines. In vitro studies showed IgE engagement with FcεRI and CD23 and Bet v 1-dependent degranulation. Overall, we hereby present fully functional, human IgE, IgG1, and IgG4 sharing the same variable region against Bet v 1 and showcase possible applications in first mechanistic studies. Furthermore, our IgG antibodies might be useful candidates for passive immunotherapy of birch pollen allergy.}, author = {Köhler, Verena K. and Crescioli, Silvia and Fazekas-Singer, Judit and Bax, Heather J. and Hofer, Gerhard and Pranger, Christina L. and Hufnagl, Karin and Bianchini, Rodolfo and Flicker, Sabine and Keller, Walter and Karagiannis, Sophia N. and Jensen-Jarolim, Erika}, issn = {1422-0067}, journal = {International Journal of Molecular Sciences}, number = {16}, publisher = {MDPI}, title = {{Filling the antibody pipeline in allergy: PIPE cloning of IgE, IgG1 and IgG4 against the major birch pollen allergen Bet v 1}}, doi = {10.3390/ijms21165693}, volume = {21}, year = {2020}, } @article{8226, author = {Gotovina, Jelena and Bianchini, Rodolfo and Fazekas-Singer, Judit and Herrmann, Ina and Pellizzari, Giulia and Haidl, Ian D. and Hufnagl, Karin and Karagiannis, Sophia N. and Marshall, Jean S. and Jensen‐Jarolim, Erika}, issn = {0105-4538}, journal = {Allergy}, publisher = {Wiley}, title = {{Epinephrine drives human M2a allergic macrophages to a regulatory phenotype reducing mast cell degranulation in vitro}}, doi = {10.1111/all.14299}, year = {2020}, } @article{8227, author = {Ilieva, Kristina M. and Fazekas-Singer, Judit and Bax, Heather J. and Crescioli, Silvia and Montero‐Morales, Laura and Mele, Silvia and Sow, Heng Sheng and Stavraka, Chara and Josephs, Debra H. and Spicer, James F. and Steinkellner, Herta and Jensen‐Jarolim, Erika and Tutt, Andrew N. J. and Karagiannis, Sophia N.}, issn = {0105-4538}, journal = {Allergy}, number = {10}, pages = {1985--1989}, publisher = {Wiley}, title = {{AllergoOncology: Expression platform development and functional profiling of an anti‐HER2 IgE antibody}}, doi = {10.1111/all.13818}, volume = {74}, year = {2019}, } @article{8228, abstract = {Background: Atopics have a lower risk for malignancies, and IgE targeted to tumors is superior to IgG in fighting cancer. Whether IgE-mediated innate or adaptive immune surveillance can confer protection against tumors remains unclear. Objective: We aimed to investigate the effects of active and passive immunotherapy to the tumor-associated antigen HER-2 in three murine models differing in Epsilon-B-cell-receptor expression affecting the levels of expressed IgE. Methods: We compared the levels of several serum specific anti-HER-2 antibodies (IgE, IgG1, IgG2a, IgG2b, IgA) and the survival rates in low-IgE ΔM1M2 mice lacking the transmembrane/cytoplasmic domain of Epsilon-B-cell-receptors expressing reduced IgE levels, high-IgE KN1 mice expressing chimeric Epsilon-Gamma1-B-cell receptors with 4-6-fold elevated serum IgE levels, and wild type (WT) BALB/c. Prior engrafting mice with D2F2/E2 mammary tumors overexpressing HER-2, mice were vaccinated with HER-2 or vehicle control PBS using the Th2-adjuvant Al(OH)3 (active immunotherapy), or treated with the murine anti-HER-2 IgG1 antibody 4D5 (passive immunotherapy). Results: Overall, among the three strains of mice, HER-2 vaccination induced significantly higher levels of HER-2 specific IgE and IgG1 in high-IgE KN1, while low-IgE ΔM1M2 mice had higher IgG2a levels. HER-2 vaccination and passive immunotherapy prolonged the survival in tumor-grafted WT and low-IgE ΔM1M2 strains compared with treatment controls; active vaccination provided the highest benefit. Notably, untreated high-IgE KN1 mice displayed the longest survival of all strains, which could not be further extended by active or passive immunotherapy. Conclusion: Active and passive immunotherapies prolong survival in wild type and low-IgE ΔM1M2 mice engrafted with mammary tumors. High-IgE KN1 mice have an innate survival benefit following tumor challenge.}, author = {Singer, Josef and Achatz-Straussberger, Gertrude and Bentley-Lukschal, Anna and Fazekas-Singer, Judit and Achatz, Gernot and Karagiannis, Sophia N. and Jensen-Jarolim, Erika}, issn = {1939-4551}, journal = {World Allergy Organization Journal}, number = {7}, publisher = {Elsevier}, title = {{AllergoOncology: High innate IgE levels are decisive for the survival of cancer-bearing mice}}, doi = {10.1016/j.waojou.2019.100044}, volume = {12}, year = {2019}, } @article{8229, abstract = {Food proteins may get nitrated by various exogenous or endogenous mechanisms. As individuals might get recurrently exposed to nitrated proteins via daily diet, we aimed to investigate the effect of repeatedly ingested nitrated food proteins on the subsequent immune response in non-allergic and allergic mice using the milk allergen beta-lactoglobulin (BLG) as model food protein in a mouse model. Evaluating the presence of nitrated proteins in food, we could detect 3-nitrotyrosine (3-NT) in extracts of different foods and in stomach content extracts of non-allergic mice under physiological conditions. Chemically nitrated BLG (BLGn) exhibited enhanced susceptibility to degradation in simulated gastric fluid experiments compared to untreated BLG (BLGu). Gavage of BLGn to non-allergic animals increased interferon-γ and interleukin-10 release of stimulated spleen cells and led to the formation of BLG-specific serum IgA. Allergic mice receiving three oral gavages of BLGn had higher levels of mouse mast cell protease-1 (mMCP-1) compared to allergic mice receiving BLGu. Regardless of the preceding immune status, non-allergic or allergic, repeatedly ingested nitrated food proteins seem to considerably influence the subsequent immune response.}, author = {Ondracek, Anna S. and Heiden, Denise and Oostingh, Gertie J. and Fuerst, Elisabeth and Fazekas-Singer, Judit and Bergmayr, Cornelia and Rohrhofer, Johanna and Jensen-Jarolim, Erika and Duschl, Albert and Untersmayr, Eva}, issn = {2072-6643}, journal = {Nutrients}, number = {10}, publisher = {MDPI}, title = {{Immune effects of the nitrated food allergen beta-lactoglobulin in an experimental food allergy model}}, doi = {10.3390/nu11102463}, volume = {11}, year = {2019}, } @article{8231, author = {Fazekas-Singer, Judit and Singer, Josef and Ilieva, Kristina M. and Matz, Miroslawa and Herrmann, Ina and Spillner, Edzard and Karagiannis, Sophia N. and Jensen-Jarolim, Erika}, issn = {0091-6749}, journal = {Journal of Allergy and Clinical Immunology}, number = {3}, pages = {973--976.e11}, publisher = {Elsevier}, title = {{AllergoOncology: Generating a canine anticancer IgE against the epidermal growth factor receptor}}, doi = {10.1016/j.jaci.2018.04.021}, volume = {142}, year = {2018}, } @article{8232, abstract = {Anti-epidermal growth factor receptor (EGFR) antibody therapy is used in EGFR expressing cancers including lung, colon, head and neck, and bladder cancers, however results have been modest. Near infrared photoimmunotherapy (NIR-PIT) is a highly selective tumor treatment that employs an antibody-photo-absorber conjugate which is activated by NIR light. NIR-PIT is in clinical trials in patients with recurrent head and neck cancers using cetuximab-IR700 as the conjugate. However, its use has otherwise been restricted to mouse models. This is an effort to explore larger animal models with NIR-PIT. We describe the use of a recombinant canine anti-EGFR monoclonal antibody (mAb), can225IgG, conjugated to the photo-absorber, IR700DX, in three EGFR expressing canine transitional cell carcinoma (TCC) cell lines as a prelude to possible canine clinical studies. Can225-IR700 conjugate showed specific binding and cell-specific killing after NIR-PIT on EGFR expressing cells in vitro. In the in vivo study, can225-IR700 conjugate demonstrated accumulation of the fluorescent conjugate with high tumor-to-background ratio. Tumor-bearing mice were separated into 4 groups: (1) no treatment; (2) 100 μg of can225-IR700 i.v. only; (3) NIR light exposure only; (4) 100 μg of can225-IR700 i.v., NIR light exposure. Tumor growth was significantly inhibited by NIR-PIT treatment compared with the other groups (p < 0.001), and significantly prolonged survival was achieved (p < 0.001 vs. other groups) in the treatment groups. In conclusion, NIR-PIT with can225-IR700 is a promising treatment for canine EGFR-expressing cancers, including invasive transitional cell carcinoma in pet dogs, that could provide a pathway to translation to humans.}, author = {Nagaya, Tadanobu and Okuyama, Shuhei and Ogata, Fusa and Maruoka, Yasuhiro and Knapp, Deborah W. and Karagiannis, Sophia N. and Fazekas-Singer, Judit and Choyke, Peter L. and LeBlanc, Amy K. and Jensen-Jarolim, Erika and Kobayashi, Hisataka}, issn = {1949-2553}, journal = {Oncotarget}, pages = {19026--19038}, publisher = {Impact Journals}, title = {{Near infrared photoimmunotherapy targeting bladder cancer with a canine anti-epidermal growth factor receptor (EGFR) antibody}}, doi = {10.18632/oncotarget.24876}, volume = {9}, year = {2018}, } @article{8233, abstract = {The M2a subtype of macrophages plays an important role in human immunoglobulin E (IgE-mediated allergies) and other Th2 type immune reactions. In contrast, very little is known about these cells in the dog. Here we describe an in vitro method to activate canine histiocytic DH82 cells and primary canine monocyte-derived macrophages (MDMs) toward the M2a macrophages using human cytokines. For a side-by-side comparison, we compared the canine cells to human MDMs, and the human monocytic cell line U937 activated towards M1 and M2a cells on the cellular and molecular level. In analogy to activated human M2a cells, canine M2a, differentiated from both DH82 and MDMs, showed an increase in CD206 surface receptor expression compared to M1. Interestingly, canine M2a, but not M1 derived from MDM, upregulated the high-affinity IgE receptor (FcεRI). Transcription levels of M2a-associated genes (IL10, CCL22, TGFβ, CD163) showed a diverse pattern between the human and dog species, whereas M1 genes (IDO1, CXCL11, IL6, TNF-α) were similarly upregulated in canine and human M1 cells (cell lines and MDMs). We suggest that our novel in vitro method will be suitable in comparative allergology studies focussing on macrophages.}, author = {Herrmann, Ina and Gotovina, Jelena and Fazekas-Singer, Judit and Fischer, Michael B. and Hufnagl, Karin and Bianchini, Rodolfo and Jensen-Jarolim, Erika}, issn = {0145-305X}, journal = {Developmental & Comparative Immunology}, number = {5}, pages = {118--127}, publisher = {Elsevier}, title = {{Canine macrophages can like human macrophages be in vitro activated toward the M2a subtype relevant in allergy}}, doi = {10.1016/j.dci.2018.01.005}, volume = {82}, year = {2018}, } @article{8234, abstract = {Molecular imaging probes such as PET-tracers have the potential to improve the accuracy of tumor characterization by directly visualizing the biochemical situation. Thus, molecular changes can be detected early before morphological manifestation. The A3 adenosine receptor (A3AR) is described to be highly expressed in colon cancer cell lines and human colorectal cancer (CRC), suggesting this receptor as a tumor marker. The aim of this preclinical study was the evaluation of FE@SUPPY as a PET-tracer for CRC using in vitro imaging and in vivo PET imaging. First, affinity and selectivity of FE@SUPPY and its metabolites were determined, proving the favorable binding profile of FE@SUPPY. The human adenocarcinoma cell line HT-29 was characterized regarding its hA3AR expression and was subsequently chosen as tumor graft. Promising results regarding the potential of FE@SUPPY as a PET-tracer for CRC imaging were obtained by autoradiography as ≥2.3-fold higher accumulation of FE@SUPPY was found in CRC tissue compared to adjacent healthy colon tissue from the same patient. Nevertheless, first in vivo studies using HT-29 xenografts showed insufficient tumor uptake due to (1) poor conservation of target expression in xenografts and (2) unfavorable pharmacokinetics of FE@SUPPY in mice. We therefore conclude that HT-29 xenografts are not adequate to visualize hA3ARs using FE@SUPPY.}, author = {Balber, T. and Singer, Judit and Berroterán-Infante, N. and Dumanic, M. and Fetty, L. and Fazekas-Singer, J. and Vraka, C. and Nics, L. and Bergmann, M. and Pallitsch, K. and Spreitzer, H. and Wadsak, W. and Hacker, M. and Jensen-Jarolim, E. and Viernstein, H. and Mitterhauser, M.}, issn = {1555-4309}, journal = {Contrast Media & Molecular Imaging}, publisher = {Hindawi}, title = {{Preclinical in vitro and in vivo evaluation of [18F]FE@SUPPY for cancer PET imaging: Limitations of a xenograft model for colorectal cancer}}, doi = {10.1155/2018/1269830}, volume = {2018}, year = {2018}, } @article{8274, abstract = {Background/Aim: Our aim was to investigate the crosstalk between tumor and immune cells (M2 macrophages) and its effects on cyclo-oxygenase-2 (COX2) regulation in canine mammary tumors (CMT). Materials and Methods: Sh1b CMT cells and human BT474 mammary or HT29 colon cancer cells were co-cultured with canine peripheral blood mononuclear cells (PBMCs) or with macrophage-like differentiated THP1 monocytes (dTHP1). Intracellular COX2 expression by PBMCs, dTHP1 and cancer cells was evaluated by flow cytometry. Results: Co-culturing of Sh1b and canine PBMCs induced COX2 overexpression in CMT cells. In turn, COX2 expression by PBMCs, mostly CD68+ macrophages, was attenuated by co-culture with Sh1b (p=0.0001). In accordance, co-culture with dTHP1 prompted intracellular production of COX2 in both Sh1b CMT cells and HT29 human colon cancer cells and reduced production of COX2 in BT474 human mammary cancer cells. The intracellular COX2 expression from dTHP1 decreased when treated with conditioned medium from cultured Sh1b and HT29 cancer cells. Conclusion: Bidirectional COX2 regulation between cancer and monocytes/macrophages might shape a tolerogenic tumor microenvironment in CMT.}, author = {Carvalho, Maria Isabel and Bianchini, Rodolfo and Fazekas-Singer, Judit and Herrmann, Ina and Flickinger, Irene and Thalhammer, Johann G. and Pires, Isabel and Jensen-Jarolim, Erika and Queiroga, Felisbina L.}, issn = {1791-7530}, journal = {Anticancer Research}, number = {5}, pages = {2811--2817}, publisher = {International Institute of Anticancer Research}, title = {{Bidirectional regulation of COX-2 expression between cancer cells and macrophages}}, doi = {10.21873/anticanres.12525}, volume = {38}, year = {2018}, } @article{8235, abstract = {Due to large homology of human and canine EGFR, dogs suffering from spontaneous EGFR+ cancer can be considered as ideal translational models. Thereby, novel immunotherapeutic compounds can be developed for both human and veterinary patients. This study describes the radiolabeling of a canine anti-EGFR IgG antibody (can225IgG) with potential diagnostic and therapeutic value in comparative clinical settings. Can225IgG was functionalized with DTPA for subsequent chelation with the radionuclide 99mTc. Successful coupling of 10 DTPA molecules per antibody on average was proven by significant mass increase in MALDI-TOF spectroscopy, gel electrophoresis and immunoblots. Following functionalization and radiolabeling, 99mTc-DTPA-can225IgG fully retained its binding capacity towards human and canine EGFR in flow cytometry, immuno- and radioblots, and autoradiography. The affinity of radiolabeled can225IgG was determined to KD 0.8 ±0.0031 nM in a real-time kinetics assay on canine carcinoma cells by a competition binding technique. Stability tests of the radiolabeled compound identified TRIS buffered saline as the ideal formulation for short-term storage with 87.11 ±6.04% intact compound being still detected 60 minutes post radiolabeling. High stability, specificity and EGFR binding affinity pinpoint towards 99mTc-radiolabeled can225IgG antibody as an ideal lead compound for the first proof-of-concept diagnostic and therapeutic applications in canine cancer patients.}, author = {Fazekas-Singer, Judit and Berroterán-Infante, Neydher and Rami-Mark, Christina and Dumanic, Monika and Matz, Miroslawa and Willmann, Michael and Andreae, Fritz and Singer, Josef and Wadsak, Wolfgang and Mitterhauser, Markus and Jensen-Jarolim, Erika}, issn = {1949-2553}, journal = {Oncotarget}, pages = {83128--83141}, publisher = {Impact Journals}, title = {{Development of a radiolabeled caninized anti-EGFR antibody for comparative oncology trials}}, doi = {10.18632/oncotarget.20914}, volume = {8}, year = {2017}, } @article{8236, abstract = {Th2 immunity and allergic immune surveillance play critical roles in host responses to pathogens, parasites and allergens. Numerous studies have reported significant links between Th2 responses and cancer, including insights into the functions of IgE antibodies and associated effector cells in both antitumour immune surveillance and therapy. The interdisciplinary field of AllergoOncology was given Task Force status by the European Academy of Allergy and Clinical Immunology in 2014. Affiliated expert groups focus on the interface between allergic responses and cancer, applied to immune surveillance, immunomodulation and the functions of IgE‐mediated immune responses against cancer, to derive novel insights into more effective treatments. Coincident with rapid expansion in clinical application of cancer immunotherapies, here we review the current state‐of‐the‐art and future translational opportunities, as well as challenges in this relatively new field. Recent developments include improved understanding of Th2 antibodies, intratumoral innate allergy effector cells and mediators, IgE‐mediated tumour antigen cross‐presentation by dendritic cells, as well as immunotherapeutic strategies such as vaccines and recombinant antibodies, and finally, the management of allergy in daily clinical oncology. Shedding light on the crosstalk between allergic response and cancer is paving the way for new avenues of treatment.}, author = {Jensen-Jarolim, E. and Bax, H. J. and Bianchini, R. and Capron, M. and Corrigan, C. and Castells, M. and Dombrowicz, D. and Daniels-Wells, T. R. and Fazekas, Judit and Fiebiger, E. and Gatault, S. and Gould, H. J. and Janda, J. and Josephs, D. H. and Karagiannis, P. and Levi-Schaffer, F. and Meshcheryakova, A. and Mechtcheriakova, D. and Mekori, Y. and Mungenast, F. and Nigro, E. A. and Penichet, M. L. and Redegeld, F. and Saul, L. and Singer, J. and Spicer, J. F. and Siccardi, A. G. and Spillner, E. and Turner, M. C. and Untersmayr, E. and Vangelista, L. and Karagiannis, S. N.}, issn = {0105-4538}, journal = {Allergy}, number = {6}, pages = {866--887}, publisher = {Wiley}, title = {{AllergoOncology - the impact of allergy in oncology: EAACI position paper}}, doi = {10.1111/all.13119}, volume = {72}, year = {2017}, } @article{8237, abstract = {Monoclonal antibodies find broad application as therapy for various types of cancer by employing multiple mechanisms of action against tumors. Manipulating the Fc-mediated functions of antibodies that engage immune effector cells, such as NK cells, represents a strategy to influence effector cell activation and to enhance antibody potency and potentially efficacy. We developed a novel approach to generate and ascertain the functional attributes of Fc mutant monoclonal antibodies. This entailed coupling single expression vector (pVitro1) antibody cloning, using polymerase incomplete primer extension (PIPE) polymerase chain reaction, together with simultaneous Fc region point mutagenesis and high yield transient expression in human mammalian cells. Employing this, we engineered wild type, low (N297Q, NQ), and high (S239D/I332E, DE) FcR-binding Fc mutant monoclonal antibody panels recognizing two cancer antigens, HER2/neu and chondroitin sulfate proteoglycan 4. Antibodies were generated with universal mutagenic primers applicable to any IgG1 pVitro1 constructs, with high mutagenesis and transfection efficiency, in small culture volumes, at high yields and within 12 days from design to purified material. Antibody variants conserved their Fab-mediated recognition of target antigens and their direct anti-proliferative effects against cancer cells. Fc mutations had a significant impact on antibody interactions with Fc receptors (FcRs) on human NK cells, and consequently on the potency of NK cell activation, quantified by immune complex-mediated calcium mobilization and by antibody-dependent cellular cytotoxicity (ADCC) of tumor cells. This strategy for manipulation and testing of Fc region engagement with cognate FcRs can facilitate the design of antibodies with defined effector functions and potentially enhanced efficacy against tumor cells.}, author = {Ilieva, Kristina M. and Fazekas-Singer, Judit and Achkova, Daniela Y. and Dodev, Tihomir S. and Mele, Silvia and Crescioli, Silvia and Bax, Heather J. and Cheung, Anthony and Karagiannis, Panagiotis and Correa, Isabel and Figini, Mariangela and Marlow, Rebecca and Josephs, Debra H. and Beavil, Andrew J. and Maher, John and Spicer, James F. and Jensen-Jarolim, Erika and Tutt, Andrew N. and Karagiannis, Sophia N.}, issn = {1664-3224}, journal = {Frontiers in Immunology}, publisher = {Frontiers}, title = {{Functionally active Fc mutant antibodies recognizing cancer antigens generated rapidly at high yields}}, doi = {10.3389/fimmu.2017.01112}, volume = {8}, year = {2017}, } @article{8239, abstract = {Acrolein, a highly reactive unsaturated aldehyde, is generated in large amounts during smoking and is best known for its genotoxic capacity. Here, we aimed to assess whether acrolein at concentrations relevant for smokers may also exert immunomodulatory effects that could be relevant in allergy or cancer. In a BALB/c allergy model repeated nasal exposure to acrolein abrogated allergen-specific antibody and cytokine formation, and led to a relative accumulation of regulatory T cells in the lungs. Only the acrolein-treated mice were protected from bronchial hyperreactivity as well as from anaphylactic reactions upon challenge with the specific allergen. Moreover, grafted D2F2 tumor cells grew faster and intratumoral Foxp3+ cell accumulation was observed in these mice compared to sham-treated controls. Results from reporter cell lines suggested that acrolein acts via the aryl-hydrocarbon receptor which could be inhibited by resveratrol and 3′-methoxy-4′-nitroflavone Acrolein- stimulation of human PBMCs increased Foxp3+ expression by T cells which could be antagonized by resveratrol. Our mouse and human data thus revealed that acrolein exerts systemic immunosuppression by promoting Foxp3+ regulatory cells. This provides a novel explanation why smokers have a lower allergy, but higher cancer risk.}, author = {Roth-Walter, Franziska and Bergmayr, Cornelia and Meitz, Sarah and Buchleitner, Stefan and Stremnitzer, Caroline and Fazekas, Judit and Moskovskich, Anna and Müller, Mario A. and Roth, Georg A. and Manzano-Szalai, Krisztina and Dvorak, Zdenek and Neunkirchner, Alina and Jensen-Jarolim, Erika}, issn = {2045-2322}, journal = {Scientific Reports}, publisher = {Springer Nature}, title = {{Janus-faced Acrolein prevents allergy but accelerates tumor growth by promoting immunoregulatory Foxp3+ cells: Mouse model for passive respiratory exposure}}, doi = {10.1038/srep45067}, volume = {7}, year = {2017}, } @article{8240, abstract = {Background/Aim: Cancer cell lines are indispensible surrogate models in cancer research, as they can be used off-the-shelf, expanded to the desired extent, easily modified and exchanged between research groups for affirmation, reproduction or follow-up experiments. As malignant cells are prone to genomic instability, phenotypical changes may occur after certain passages in culture. Thus, cell lines have to be regularly authenticated to ensure data quality. In between experiments these cell lines are often stored in liquid nitrogen for extended time periods. Although freezing of cells is a necessary evil, little research is performed on how long-term storage affects cancer cell lines. Therefore, this study investigated the effects of a 28-year long liquid nitrogen storage period on BT474 cells with regard to phenotypical changes, differences in cell-surface receptor expression as well as cytokine and gene expressional variations. Methods: Two batches of BT474 cells, one frozen in 1986, the other directly purchased from ATCC were investigated by light microscopy, cell growth analysis, flow cytometry and cytokine as well as whole-transcriptome expression profiling. Results: The cell lines were morphologically indifferent and showed similar growth rates and similar cell-surface receptor expression. Transcriptome analysis revealed significant differences in only 26 of 40,716 investigated RefSeq transcripts with 4 of them being up-regulated and 22 down-regulated. Conclusion: This study demonstrates that even after very long periods of storage in liquid nitrogen, cancer cell lines display only minimal changes in their gene expression profiles. However, also such minor changes should be carefully assessed before continuation of experiments, especially if phenotypic alterations can be additionally observed.}, author = {Fazekas, Judit and Grunt, Thomas W. and Jensen-Jarolim, Erika and Singer, Josef}, issn = {1949-2553}, journal = {Oncotarget}, pages = {35076--35087}, publisher = {Impact Journals}, title = {{Long term storage in liquid nitrogen leads to only minor phenotypic and gene expression changes in the mammary carcinoma model cell line BT474}}, doi = {10.18632/oncotarget.16623}, volume = {8}, year = {2017}, } @article{8241, abstract = {Background: Anticancer vaccines could represent a valuable complementary strategy to established therapies, especially in settings of early stage and minimal residual disease. HER-2 is an important target for immunotherapy and addressed by the monoclonal antibody trastuzumab. We have previously generated HER-2 mimotope peptides from phage display libraries. The synthesized peptides were coupled to carriers and applied for epitope-specific induction of trastuzumab-like IgG. For simplification and to avoid methodological limitations of synthesis and coupling chemistry, we herewith present a novel and optimized approach by using adeno-associated viruses (AAV) as effective and high-density mimotope-display system, which can be directly used for vaccination. Methods: An AAV capsid display library was constructed by genetically incorporating random peptides in a plasmid encoding the wild-type AAV2 capsid protein. AAV clones, expressing peptides specifically reactive to trastuzumab, were employed to immunize BALB/c mice. Antibody titers against human HER-2 were determined, and the isotype composition and functional properties of these were tested. Finally, prophylactically immunized mice were challenged with human HER-2 transfected mouse D2F2/E2 cells. Results: HER-2 mimotope AAV-vaccines induced antibodies specific to human HER-2. Two clones were selected for immunization of mice, which were subsequently grafted D2F2/E2 cells. Both mimotope AAV clones delayed the growth of tumors significantly, as compared to controls. Conclusion: In this study, a novel mimotope AAV-based platform was created allowing the isolation of mimotopes, which can be directly used as anticancer vaccines. The example of trastuzumab AAV-mimotopes demonstrates that this vaccine strategy could help to establish active immunotherapy for breast-cancer patients.}, author = {Singer, Josef and Manzano-Szalai, Krisztina and Fazekas, Judit and Thell, Kathrin and Bentley-Lukschal, Anna and Stremnitzer, Caroline and Roth-Walter, Franziska and Weghofer, Margit and Ritter, Mirko and Pino Tossi, Kerstin and Hörer, Markus and Michaelis, Uwe and Jensen-Jarolim, Erika}, issn = {2162-402X}, journal = {OncoImmunology}, number = {7}, publisher = {Taylor & Francis}, title = {{Proof of concept study with an HER-2 mimotope anticancer vaccine deduced from a novel AAV-mimotope library platform}}, doi = {10.1080/2162402x.2016.1171446}, volume = {5}, year = {2016}, } @article{8242, author = {Einhorn, Lukas and Fazekas, Judit and Muhr, Martina and Schoos, Alexandra and Oida, Kumiko and Singer, Josef and Panakova, Lucia and Manzano-Szalai, Krisztina and Jensen-Jarolim, Erika}, issn = {0091-6749}, journal = {Journal of Allergy and Clinical Immunology}, number = {2}, publisher = {Elsevier}, title = {{Generation of recombinant FcεRIα of dog, cat and horse for component-resolved allergy diagnosis in veterinary patients}}, doi = {10.1016/j.jaci.2014.12.1263}, volume = {135}, year = {2015}, } @article{8244, abstract = {Passive immunotherapy with monoclonal antibodies represents a cornerstone of human anticancer therapies, but has not been established in veterinary medicine yet. As the tumor-associated antigen EGFR (ErbB-1) is highly conserved between humans and dogs, and considering the effectiveness of the anti-EGFR antibody cetuximab in human clinical oncology, we present here a “caninized” version of this antibody, can225IgG, for comparative oncology studies. Variable region genes of 225, the murine precursor of cetuximab, were fused with canine constant heavy gamma and kappa chain genes, respectively, and transfected into Chinese hamster ovary (CHO) DUKX-B11 cells. Of note, 480 clones were screened and the best clones were selected according to productivity and highest specificity in EGFR-coated ELISA. Upon purification with Protein G, the recombinant cetuximab-like canine IgG was tested for integrity, correct assembly, and functionality. Specific binding to the surface of EGFR-overexpressing cells was assessed by flow cytometry and immunofluorescence; moreover, binding to canine mammary tissue was demonstrated by immunohistochemistry. In cell viability and proliferation assays, incubation with can225IgG led to significant tumor cell growth inhibition. Moreover, this antibody mediated significant tumor cell killing via phagocytosis in vitro. We thus present here, for the first time, the generation of a canine IgG antibody and its hypothetical structure. On the basis of its cetuximab-like binding site, on the one hand, and the expression of a 91% homologous EGFR molecule in canine cancer, on the other hand, this antibody may be a promising research compound to establish passive immunotherapy in dog patients with cancer.}, author = {Singer, J. and Fazekas, Judit and Wang, W. and Weichselbaumer, M. and Matz, M. and Mader, A. and Steinfellner, W. and Meitz, S. and Mechtcheriakova, D. and Sobanov, Y. and Willmann, M. and Stockner, T. and Spillner, E. and Kunert, R. and Jensen-Jarolim, E.}, issn = {1535-7163}, journal = {Molecular Cancer Therapeutics}, number = {7}, pages = {1777--1790}, publisher = {American Association for Cancer Research}, title = {{Generation of a canine anti-EGFR (ErbB-1) antibody for passive immunotherapy in dog cancer patients}}, doi = {10.1158/1535-7163.mct-13-0288}, volume = {13}, year = {2014}, }