TY - CHAP A1 - Neumann-Schaal, M. A1 - Messerschmidt, Katrin A1 - Grenz, N. A1 - Micheel, Burkhard A1 - Heilmann, K. T1 - Use of antibody gene library for the isolation of specific single chain antibodies by ampicillin-antigen conjugates T2 - Immunology : an official journal of the British Society for Immunology Y1 - 2012 SN - 0019-2805 VL - 137 IS - 3 SP - 661 EP - 661 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - VanderVen, Peter F. M. A1 - Ehler, Elisabeth A1 - Vakeel, Padmanabhan A1 - Eulitz, Stefan A1 - Schenk, Jörg A. A1 - Milting, Hendrik A1 - Micheel, Burkhard A1 - Fürst, Dieter Oswald T1 - Unusual splicing events result in distinct Xin isoforms that associate differentially with filamin c and Mena/ VASP N2 - Filamin c is the predominantly expressed filamin isoform in striated muscles. It is localized in myofibrillar Z- discs, where it binds FATZ and myotilin, and in myotendinous junctions and intercalated discs. Here, we identify Xin, the protein encoded by the human gene 'cardiomyopathy associated 1' (CMYA1) as filamin c binding partner at these specialized structures where the ends of myofibrils are attached to the sarcolemma. Xin directly binds the EVH1 domain proteins Mena and VASP. In the adult heart, Xin and Mena/VASP colocalize with filamin c in intercalated discs. In cultured cardiomyocytes, the proteins also localize in the nonstriated part of myofibrils, where sarcomeres are assembled and an extensive reorganization of the actin cytoskeleton occurs. Unusual intraexonic splicing events result in the existence of three Xin isoforms that associate differentially with its ligands. The identification of the complex filamin c-Xin-Mena/VASP provides a first glance on the role of Xin in the molecular mechanisms involved in developmental and adaptive remodeling of the actin cytoskeleton during cardiac morphogenesis and sarcomere assembly. (c) 2006 Elsevier Inc. All rights reserved Y1 - 2006 U6 - https://doi.org/10.1016/j.yexcr.2006.03.015 ER - TY - JOUR A1 - Bier, Frank Fabian A1 - Ehrentreich-Förster, Eva A1 - Scheller, Frieder W. A1 - Makower, Alexander A1 - Eremenko, A. V. A1 - Wollenberger, Ursula A1 - Bauer, Christian G. A1 - Pfeiffer, Dorothea A1 - Micheel, Burkhard T1 - Ultrasensitive biosensors Y1 - 1996 ER - TY - JOUR A1 - Micheel, Burkhard T1 - Tumorantigene und ihre Nutzung für eine Therapie mit Antikörpern Y1 - 1998 ER - TY - JOUR A1 - Engel, Robert A1 - Micheel, Burkhard A1 - Hanack, Katja T1 - Three-dimensional cell culture approach for in vitro immunization and the production of monoclonal antibodies JF - Biomedical materials : materials for tissue engineering and regenerative medicine N2 - The generation of monoclonal antibodies using an in vitro immunization approach is a promising alternative to conventional hybridoma technology. As recently published, the in vitro approach enables an antigen-specific activation of B lymphocytes within 10-12 d followed by immortalization and subsequent selection of hybridomas. This in vitro process can be further improved by using a three-dimensional surrounding to stabilize the complex microenvironment required for a successful immune reaction. In this study, the suitability of Geltrex as a material for the generation of monoclonal antigen-specific antibodies by in vitro immunization was analyzed. We could show that dendritic cells, B cells, and T cells were able to travel through and interact inside of the matrix, leading to the antigen-specific activation of T and B cells. For cell recovery and subsequent hybridoma technique the suitability of dispase and Corning cell recovery solution (CRS) was compared. In our experiments, the use of dispase resulted in a severe alteration of cell surface receptor expression patterns and significantly higher cell death, while we could not detect an adverse effect of Corning CRS. Finally, an easy approach for high-density cell culture was established by printing an alginate ring inside a cell culture vessel. The ring was filled with Geltrex, cells, and medium to ensure a sufficient supply during cultivation. Using this approach, we were able to generate monoclonal hybridomas that produce antigen-specific antibodies against ovalbumin and the SARS-CoV-2 nucleocapsid protein. KW - monoclonal antibody KW - hybridoma technology KW - in vitro immunization KW - 3D KW - cell culture KW - Geltrex Y1 - 2022 U6 - https://doi.org/10.1088/1748-605X/ac7b00 SN - 1748-6041 SN - 1748-605X VL - 17 IS - 5 PB - Inst. of Physics CY - London ER - TY - JOUR A1 - Schlag, Peter M. A1 - Osterziel, Karl Joseph A1 - Özcelik, Cemil A1 - Scherneck, Siegfried A1 - Wenzel, Katrin A1 - Daskalow, Katjana A1 - Herse, Florian A1 - Seitz, Susanne A1 - Zacharias, Ute A1 - Schenk, Jörg A. A1 - Schulz, Herbert A1 - Hübner, Norbert A1 - Micheel, Burkhard T1 - The protein phosphatase 1 inhibitor KEPI is down regulated in breast cancer cell lines and tissues and involved in the regulation of the tumour suppressor EGR1 via the MEK-ERK pathway N2 - KEPI is a protein kinase C-potentiated inhibitory protein for type 1 Ser/Thr protein phosphatases. We found no or reduced expression of KEPI in breast cancer cell lines, breast tumors and metastases in comparison to normal breast cell lines and tissues, respectively. KEPI protein expression and ubiquitous localization was detected with a newly generated antibody. Ectopic KEPI expression in MCF7 breast cancer cells induced differential expression of 95 genes, including the up-regulation of the tumor suppressors EGR1 (early growth response 1) and PTEN (phosphatase and tensin homolog), which is regulated by EGR1. We further show that the up-regulation of EGR1 in MCF7/KEPI cells is mediated by MEK-ERK signaling. The inhibition of this pathway by the MEK inhibitor UO126 led to a strong decrease in EGR1 expression in MCF7/KEPI cells. These results reveal a novel role for KEPI in the regulation of the tumor suppressor gene EGR1 via activation of the MEK-ERK MAPK pathway. Y1 - 2008 UR - http://www.atypon-link.com/doi/abs/10.1515/BC.2007.062 ER - TY - JOUR A1 - Heilmann, Katja A1 - Groth, Thomas A1 - Behrsing, Olaf A1 - Wagner, Albrecht A1 - Schossig-Tiedemann, Michael A1 - Lendlein, Andreas A1 - Micheel, Burkhard T1 - The influence of the chemical composition of cell culture material on the growth and antibody production of hybridoma cells N2 - The multiplication and antibody production of murine hybridoma cells cultured on five different polymer membranes were tested and compared with conventional tissue culture polystyrene (TCPS). Membranes were prepared from polyacrylonitrile (PAN) and acrylonitrile copolymerized with N-vinylpyrrolidone (NVP20, NVP30), Na-methallylsulfonate (NaMAS) and N-(3-amino-propyl-methacrylamide-hydrochloride) (APMA). Cell number and antibody concentration were quantified as criteria for viability and productivity. Adhesion of hybridoma cells was characterized by vital and scanning electron microscopy. The results suggest that a strong adhesion of cells, observed on APMA and TCPS, increased cell growth but reduced monoclonal antibody production. In contrast membranes with lowered adhesivity such as NVP20 provided favourable conditions for monoclonal antibody production. In addition it was shown that this membrane also possessed a minor fouling as indicated by the low decrease of water flux across the membrane after protein adsorption. It was concluded that NVP20 could be a suitable material for the development of hollow fibre membranes for bioreactors. Y1 - 2005 UR - http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T3C-4DPYNGY- 4&_coverDate=02%2F09%2F2005&_alid=268995355&_rdoc=1&_fmt=&_orig=search&_qd=1&_cdi=4943&_sort=d&view=c&_acct=C000053886&_v e ER - TY - JOUR A1 - Micheel, Burkhard T1 - Stichworte zu Thema Immunologie Y1 - 2000 ER - TY - JOUR A1 - Micheel, Burkhard T1 - Stichworte zu Thema Immunologie Y1 - 1999 ER - TY - JOUR A1 - Jandrig, Burkhard A1 - Seitz, Susanne A1 - Hinzmann, Bernd A1 - Arnold, Wolfgang A1 - Micheel, Burkhard A1 - Koelble, Konrad A1 - Siebert, Reiner A1 - Schwartz, Arnfried A1 - Ruecker, Karin A1 - Schlag, Peter M. A1 - Scherneck, Siegfried A1 - Rosenthal, Andra T1 - ST18 is a breast cancer tumor suppressor gene at human chromosome 8q11.2 N2 - We have identified a gene, ST18 (suppression of tumorigenicity 18, breast carcinoma, zinc-finger protein), within a frequent imbalanced region of chromosome 8q11 as a breast cancer tumor suppressor gene. The ST18 gene encodes a zinc-finger DNA-binding protein with six fingers of the C2HC type (configuration Cys-X5-Cys-X12-His-X4-Cys) and an SMC domain. ST18 has the potential to act as transcriptional regulator. ST18 is expressed in a number of normal tissues including mammary epithelial cells although the level of expression is quite low. In breast cancer cell lines and the majority of primary breast tumors, ST18 mRNA is significantly downregulated. A 160 bp region within the promoter of the ST18 gene is hypermethylated in about 80% of the breast cancer samples and in the majority of breast cancer cell lines. The strong correlation between ST18 promoter hypermethylation and loss of ST18 expression in tumor cells suggests that this epigenetic mechanism is responsible for tumor-specific downregulation. We further show that ectopic ST18 expression in MCF-7 breast cancer cells strongly inhibits colony formation in soft agar and the formation of tumors in a xenograft mouse model Y1 - 2004 UR - http://www.nature.com/cgi-taf/DynaPage.taf?file=/onc/journal/v23/n57/abs/ 1208131a.html&dynoptions=doi1113987275 ER - TY - THES A1 - Benkert, Alexander A1 - Scheller, Frieder W. A1 - Schössler, W. A1 - Micheel, Burkhard A1 - Warsinke, Axel T1 - Size exclusion redox-labeled immunoassay (SERI) : a new format for homogeneous amperometric creatinine determination Y1 - 2000 ER - TY - JOUR A1 - Stöcklein, Walter F. M. A1 - Rohde, M. A1 - Scharte, Gudrun A1 - Behrsing, Olaf A1 - Warsinke, Axel A1 - Micheel, Burkhard A1 - Scheller, Frieder W. T1 - Sensitive detection of triazine and phenylurea pesticides in pure organic solvent by enzyme linked immunsorbent assay (ELISA): stabilities, solubilities and sensitives Y1 - 2000 ER - TY - CHAP A1 - Holzlöhner, Pamela A1 - Schliebs, Erik A1 - Maier, Natalia A1 - Füner, Jonas A1 - Micheel, Burkhard A1 - Heilmann, Katja T1 - Production of monoclonal camelid antibodies by means of hybridoma technology T2 - The journal of immunology Y1 - 2013 SN - 0022-1767 VL - 190 PB - American Assoc. of Immunologists CY - Bethesda ER - TY - JOUR A1 - Stech, Marlitt A1 - Merk, Helmut A1 - Schenk, Jörg A. A1 - Stöcklein, Walter F. M. A1 - Wüstenhagen, Doreen Anja A1 - Micheel, Burkhard A1 - Duschl, Claus A1 - Bier, Frank Fabian A1 - Kubick, Stefan T1 - Production of functional antibody fragments in a vesicle-based eukaryotic cell-free translation system JF - Journal of biotechnology N2 - Cell-free protein synthesis is of increasing interest for the rapid and high-throughput synthesis of many proteins, in particular also antibody fragments. In this study, we present a novel strategy for the production of single chain antibody fragments (scFv) in a eukaryotic in vitro translation system. This strategy comprises the cell-free expression, isolation and label-free interaction analysis of a model antibody fragment synthesized in two differently prepared insect cell lysates. These lysates contain translocationally active microsomal structures derived from the endoplasmic reticulum (ER), allowing for posttranslational modifications of cell-free synthesized proteins. Both types of these insect cell lysates enable the synthesis and translocation of scFv into ER-derived vesicles. However, only the one that has a specifically adapted redox potential yields functional active antibody fragments. We have developed a new methodology for the isolation of functional target proteins based on the translocation of cell-free produced scFv into microsomal structures and subsequent collection of protein-enriched vesicles. Antibody fragments that have been released from these vesicles are shown to be well suited for label-free binding studies. Altogether, these results show the potential of insect cell lysates for the production, purification and selection of antibody fragments in an easy-to-handle and time-saving manner. KW - Cell-free KW - In vitro translation KW - Single chain antibody (scFv) KW - Insect lysate KW - Surface plasmon resonance Y1 - 2012 U6 - https://doi.org/10.1016/j.jbiotec.2012.08.020 SN - 0168-1656 VL - 164 IS - 2 SP - 220 EP - 231 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Rohde, M. A1 - Schenk, Jörg A. A1 - Heymann, Stephan A1 - Behrsing, Olaf A1 - Scharte, Gudrun A1 - Kempter, Gerhard A1 - Woller, Jochen A1 - Höhne, Wolfgang A1 - Warsinke, Axel A1 - Micheel, Burkhard T1 - Production and characterization of monoclonal antibodeis against urea derivatives Y1 - 1998 ER - TY - JOUR A1 - Messerschmidt, Katrin A1 - Degen, Janine A1 - Micheel, Burkhard T1 - Oxidoreductase activity of multifunctional monoclonal antibody B13-DE1 JF - Journal of molecular recognition : an international journal devoted to research on specific molecular recognition in chemistry, biology, biotechnology and medicine N2 - The monoclonal antibody B13-DE1 binds fluorescein, several fluorescein derivatives, and three peptide mimotopes. Our results revealed that this antibody also catalyzed the redox reaction of resazurin to resorufin, which are both structurally related to fluorescein. By using sodium sulfite as a reducing agent, the antibody B13-DE1 lowered the activation energy of this reaction. The Michaelis-Menten constant and turnover number of the catalyzed reaction were determined as 4.2 mu mol/l and 0.0056 s(-1), respectively. Because the results showed that fluorescein inhibited the catalytic activity of the antibody, we assume that the antigen-binding site and the catalytic active site are identical. KW - catalytic antibody KW - fluorescein KW - oxidoreductase KW - resazurin KW - resorufin Y1 - 2011 U6 - https://doi.org/10.1002/jmr.1136 SN - 0952-3499 VL - 24 IS - 6 SP - 930 EP - 934 PB - Wiley-Blackwell CY - Malden ER - TY - JOUR A1 - Behrsing, Olaf A1 - Micheel, Burkhard T1 - Monoklonale Antikörper : Grundlagen und ihre Bedeutung in Diagnostik und Therapie Y1 - 2008 SN - 978-3-540-69412-0 ER - TY - JOUR A1 - Micheel, Burkhard T1 - Monoklonale Antikörper Y1 - 2003 ER - TY - GEN A1 - Holzlöhner, Pamela A1 - Butze, Monique A1 - Hebel, Nicole A1 - Weschke, Daniel A1 - Schliebs, E. A1 - Naumann, F. A1 - Füner, J. A1 - Micheel, Burkhard A1 - Hanack, Katja T1 - Monoclonal mouse antibodies against PBMC subpopulations of New World camelides T2 - European journal of immunology Y1 - 2016 SN - 0014-2980 SN - 1521-4141 VL - 46 SP - 1175 EP - 1175 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Micheel, Burkhard T1 - Monoclonal Antibodies Y1 - 2006 SN - 978-3-540-44244-8 ER - TY - JOUR A1 - Werner, Deljana A1 - Behrsing, Olaf A1 - Scharte, Gudrun A1 - Woller, Jochen A1 - Steup, Martin A1 - Micheel, Burkhard T1 - Monoclonal anti-diuron antibodies prevent inhibition of photosynthesis by diuron Y1 - 2002 ER - TY - JOUR A1 - Heilmann, Katja A1 - Groth, Thomas A1 - Schossig, Michael A1 - Lendlein, Andreas A1 - Micheel, Burkhard T1 - Modulation of hybridoma cell growth and antibody production by coating cell culture material with extracellular matrix proteins N2 - The influence of coating polystyrene tissue culture plates with different proteins on murine hybridoma cell growth and antibody production was investigated. Fibronectin, collagen I, bovine serum albumin and laminin were used to coat NUNC and COSTAR cell culture plates. Cell number and antibody concentration in culture fluids were quantified as indicators for cell viability, proliferation and productivity. Adhesive behaviour, morphology, expression of surface receptors of hybridoma cells and the presence of tyrosine-phosphorylated proteins in cell lysates were characterized by cell adhesion experiments, microscopy, flow cytometry and Western Blot analysis. It was shown that coatings with fibronectin (0.2 ;g/ml) lead to a substantial improvement of cell growth by 50-70% and an increase of monoclonal antibody production by 100-120%. Collagen I coatings showed an improvement in cell growth by 30-70% and by 60% for the production of monoclonal antibodies. Coatings with BSA and laminin had minor effects on these parameters. It was found that the hybridoma cell lines used in this study did not express the ;2-chain of the ;2;1-integrin, which is responsible for binding to collagen and laminin. However, the presence of ;1- integrin on the cell surface was shown, which should enable hybridoma cells to bind fibronectin. We propose, therefore, that fibronectin adsorption to cell culture materials may be a promising approach to enhance the production of monoclonal antibodies by cultivated hybridoma cells. Y1 - 2007 UR - http://www.sciencedirect.com/science/journal/1369703X U6 - https://doi.org/10.1016/j.bej.2007.01.035 SN - 1369-703X ER - TY - JOUR A1 - Benkert, Alexander A1 - Micheel, Burkhard A1 - Schössler, W. A1 - Warsinke, Axel T1 - Mischen und gleich messen : creatinin spezifisch und einfach bestimmen mit den Size exclusion redox-labeled immunoassay Y1 - 2001 ER - TY - JOUR A1 - Vogt, Birgit A1 - Warncke, Marit A1 - Micheel, Burkhard A1 - Sheriff, Ahmed T1 - Lentiviral gene transfer of CTLA4 generates B cells with reduced costimulatory properties : brief definite report N2 - Peripheral T-cell (TC) tolerance can be induced by tolerogenic antigen-presenting cell (APC). A prerequisite is the reduction or blockade of B7 of APC. Besides dendritic cell, B cells can be used as APC. Here, we show the generation B cells with reduced B7 expression by lentiviral transduction of endoplasmic reticulum (ER)-directed CTLA4. Vectors coding for the human CTL4-Ig were used for the human B-cell line Raji. Transduction efficiency was over 90% (MOI = 3). For the murine B-cell line A20 and for primary mouse B cells, murine CTLA4 was used. We show that B cells with reduced B7 expression reduce the antigen (Ag) specific TC proliferation in vitro. B cells expressing an ER-directed CTLA4 may reduce Ag-specific immune responses. Y1 - 2009 UR - http://informahealthcare.com/loi/aut U6 - https://doi.org/10.1080/08916930902832470 SN - 0891-6934 ER - TY - JOUR A1 - Sheriff, Ahmed A1 - Vogt, B. A1 - Baumgart, Martin A1 - Montag, C. A1 - Hollenbach, B. A1 - Schenk, Jörg A. A1 - Ulrich, J. A1 - Ellias, F. A1 - Micheel, Burkhard T1 - Intracellular capture of B7 in antigen-presenting cells reduces costimulatory activity N2 - CTLA-4 gene constructs were designed to express CTLA-4 exclusively in the endoplasmic reticulum (ER). Four different CTLA-4 gene constructs were transfected into HEK 293 (human embryonic kidney) and A20 (Balb/c mouse B lymphoma) cells. All constructs contained an ER retention signal and coded for CTLA-4 expression in the ER. One of the constructs, which contained the membrane part of CTLA-4, coded for an expression both on the cell surface and in the ER. Three of the expressed CTLA-4 types (including the ER-membrane-expressed form) caused a reduced surface expression of B7 in the A20 cells. Only constructs which allow dimerization of CTLA-4 showed this effect. It is assumed that intracellular CTLA-4 bound B7 and inhibited therefore the transport of B7 to the surface. The binding obviously caused also an enhanced degradation of the complexes because both proteins showed a low concentration in the transfected cell lines. CTLA-4-transfected and B7-reduced A20 cells showed a diminished costimulating activity upon T cells. This was demonstrated by a reduced proliferation of T cells from ovalbumin-immunized Balb/c mice, incubated with ovalbumin peptide-primed CTLA-4-transfected A20 cells. Y1 - 2003 UR - http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WBK-47T23XK- 8&_coverDate=02%2F21%2F2003&_alid=268969757&_rdoc=1&_fmt=&_orig=search&_qd=1&_cdi=6713&_sort=d&view=c&_acct=C000053886&_v e ER - TY - JOUR A1 - Schenk, Jörg A. A1 - Matyssek, Franziska A1 - Micheel, Burkhard T1 - Interleukin 4 increases the antibody response against Rubisco in mice N2 - The influence of interleukin 4 (IL-4) on antibody titer in serum and spleen culture supernatant in mice immunized with spinach (Spinacia oleracea L.) Rubisco was investigated. Therefore, we boosted one mouse additionally to the antigen with recombinant mouse IL-4. We found that the Rubisco-specific antibody titer in serum as well as in spleen cell culture supernatant was significantly enhanced in the IL-4 mouse. Most of the antibodies were of the IgG1 subclass. After hybridoma generation, Rubisco-specific antibodies were found in more than 95% of the wells tested compared to about 12% of the control mouse. Y1 - 2004 ER - TY - CHAP A1 - Listek, Martin A1 - Micheel, Burkhard A1 - Heilmann, K. T1 - Insertion of artificial cell surface receptors for antigen-specific labelling of hybridoma cells T2 - Immunology : an official journal of the British Society for Immunology Y1 - 2012 SN - 0019-2805 VL - 137 SP - 651 EP - 651 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Michelchen, Sophia A1 - Micheel, Burkhard A1 - Hanack, Katja T1 - In vitro immunization approach to generate specific murine monoclonal IgG antibodies JF - Journal of immunological methods : JIM N2 - Generating a monoclonal antibody to date is a time intense process that requires immunization of laboratory animals. The transfer of the humoral immune response into in vitro settings enables a shortening of this process and circumvents the necessity of in vivo immunization. However, to orchestrate the complex interplay of dendritic cells, T and B lymphocytes in vitro is very challenging. We therefore aimed for a simplified approach focusing on the protagonist of antibody production: the B lymphocyte. We activated purified murine B lymphocytes alone in vitro by using combinations of antigen and stimuli. We were able to induce a specific antibody response within ten days of culture against a viral coat protein as model antigen. Antibodies were of both IgM and IgG subclass. The stimulated B lymphocytes were transformed into permanently antibody-producing hybridomas by cell fusion technology. We furthermore used this method to induce a specific antibody response against L. pneumophila in vitro. We thus established a useful and effective in vitro protocol to generate monoclonal antibodies. By overcoming the necessity of in vivo immunization this protocol may be the first step towards a universal strategy to generate antibodies from various species. KW - Monoclonal antibody KW - Hybridoma technology KW - In vitro immunization KW - B cell activation Y1 - 2021 U6 - https://doi.org/10.1016/j.jim.2021.113149 SN - 0022-1759 SN - 1872-7905 VL - 499 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bartel, Manuela A1 - Hartmann, Stefanie A1 - Lehmann, Karola A1 - Postel, Kai A1 - Quesada, Humberto A1 - Philipp, Eva E. R. A1 - Heilmann, Katja A1 - Micheel, Burkhard A1 - Stuckas, Heiko T1 - Identification of sperm proteins as candidate biomarkers for the analysis of reproductive isolation in Mytilus: a case study for the enkurin locus JF - Marine biology : international journal on life in oceans and coastal waters N2 - Sperm proteins of the marine sessile mussels of the Mytilus edulis species complex are models to investigate reproductive isolation and speciation. This study aimed at identifying sperm proteins and their corresponding genes. This was aided by the use of monoclonal antibodies that preferentially bind to yet unknown sperm molecules. By identifying their target molecules, this approach identified proteins with relevance to Mytilus sperm function. This procedure identified 16 proteins, for example, enkurin, laminin, porin and heat shock proteins. The potential use of these proteins as genetic markers to study reproductive isolation is exemplified by analysing the enkurin locus. Enkurin evolution is driven by purifying selection, the locus displays high levels of intraspecific variation and species-specific alleles group in distinct phylogenetic clusters. These findings characterize enkurin as informative candidate biomarker for analyses of clinal variation and differential introgression in hybrid zones, for example, to understand determinants of reproductive isolation in Baltic Mytilus populations. Y1 - 2012 U6 - https://doi.org/10.1007/s00227-012-2005-7 SN - 0025-3162 VL - 159 IS - 10 SP - 2195 EP - 2207 PB - Springer CY - New York ER - TY - JOUR A1 - Böttger, Volker A1 - Peters, L.-E. A1 - Micheel, Burkhard T1 - Identification of peptide mimotopes for the fluorescein hapten binding of monoclonal antibody B13-DE1 Y1 - 1999 ER - TY - JOUR A1 - Sellrie, Frank A1 - Warsinke, Axel A1 - Micheel, Burkhard T1 - Homogeneous indirect fluorescence quenching immunoassay for the determination of low molecular weight substances JF - Analytical & bioanalytical chemistry N2 - This paper describes the principle of a homogeneous indirect fluorescence quenching immunoassay that uses monoclonal antibodies. It is a carrier-free assay system that is performed completely in solution. The assay system was established for the determination of a low molecular weight substance (hapten), the herbicide diuron, used as a model analyte. A fluorescein-monuron conjugate together with a fluorescence-quenching monoclonal anti-fluorescein antibody and an anti-analyte antibody (here an anti-diuron/monuron monoclonal antibody) were used as central components of the assay. The fluorescein-monuron conjugate can be bound either by the anti-fluorescein monoclonal antibody or by the anti-diuron/ monuron monoclonal antibody. Due to steric hindrance, binding of both antibodies to the conjugate was not possible at the same time. By selecting the antibody concentrations appropriately, a dynamic equilibrium can be established that permits the preferential binding of the anti-diuron/monuron antibody to the conjugate, which allows the fluorescein in the conjugate to fluoresce. This equilibrium can be easily altered by adding free analyte (diuron), which competes with the conjugate to bind to the anti-diuron/monuron antibody. A reduction of anti-diuron/monuron antibody binding to the conjugate results in an increase in the binding of the anti-fluorescein antibody, which leads to a decrease in the fluorescence of the conjugate. The fluorescence is therefore a direct indicator of the state of equilibrium of the system and thus also the presence of free unconjugated analyte. The determination of an analyte based on this test principle does not require any washing steps. After the test components are mixed, the dynamic equilibrium is rapidly reached and the results can be obtained in less than 5 min by measuring the fluorescence of the fluorescein. We used this test principle for the determination of diuron, which was demonstrated for concentrations of approximately 5 nM. Y1 - 2008 UR - http://www.springerlink.com/content/n7227875454216v7/ VL - 386 IS - 2 SP - 206 EP - 210 ER - TY - JOUR A1 - Holzlöhner, Pamela A1 - Butze, Monique A1 - Maier, Natalia A1 - Hebel, Nicole A1 - Schliebs, Erik A1 - Micheel, Burkhard A1 - Fuener, Jonas A1 - Heidicke, Gabriele A1 - Hanack, Katja T1 - Generation of murine monoclonal antibodies with specificity against conventional camelid IgG1 and heavy-chain only IgG2/3 JF - Veterinary Immunology and Immunopathology N2 - Camelids possess antibodies with a conventional four-chain structure consisting of two heavy and two light chains (of subclass IgG1) but further they also generate heavy-chain only antibodies (of subclass IgG2 and 3) which are fully functional in antigen binding. In this study subclass-specific murine monoclonal antibodies specific to conventional camelid IgG1 and heavy-chain only IgG2/3 were generated and validated for the use as potent secondary detection reagents. The monoclonal antibodies are able to differentiate between all camelid IgGs, conventional four-chain camelid antibodies (of subclass IgG1) and exclusively heavy chain-only antibodies (of subclasses IgG2 and IgG3). Further these antibodies were used to detect specific immune responses after vaccination of Camelids against bovine corona- and rotavirus strains and different E.coli. and Clostridia - antigens and to identify Erysipelothrix rhusiopathiae infected animals within a herd. The described antibodies are suitable as new secondary agents for the detection of different camelid subclasses and the validation of camelid immune reactions. KW - Camelid antibodies KW - Heavy-chain only antibodies KW - Monoclonal antibodies KW - Secondary antibodies KW - Vaccination KW - Disease monitoring Y1 - 2018 U6 - https://doi.org/10.1016/j.vetimm.2018.01.006 SN - 0165-2427 SN - 1873-2534 VL - 197 SP - 1 EP - 6 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Daskalow, Katjana A1 - Boisguerin, Prisca A1 - Jandrig, Burkhard A1 - van Landeghem, Frank K. H. A1 - Volkmer, Rudolf A1 - Micheel, Burkhard A1 - Schenk, Jörg A. T1 - Generation of an antibody against the protein phosphatase 1 inhibitor KEPI and characterization of the epitope N2 - A monoclonal antibody against the potential tumor suppressor kinase-enhanced protein phosphatase 1 (PP1) inhibitor KEPI (PPP1R14C) was generated and characterized. Human KEPI was expressed in Escherichia coli and used to immunize Balb/c mice. Using hybridoma technology, one clone, G18AF8, was isolated producing antibodies which bound specifically to the KEPI protein in ELISA, immunoblotting and flow cytometry. The antibody was also successfully applied to stain KEPI protein in paraffin sections of human brain. The epitope was mapped using peptide array technology and confirmed as GARVFFQSPR. This corresponds to the N-terminal region of KEPI. Amino acid substitution analysis revealed that two residues, F and Q, are essential for binding. Affinity of binding was determined by competitive ELISA as 1 mu M. In Western blot assays testing G18AF8 antibody on brain samples of several species, reactivity with hamster, rat and chicken samples was found, suggesting a broad homology of this KEPI epitope in vertebrates. This antibody could be used in expression studies at the protein level e.g. in tumor tissues. Y1 - 2010 UR - http://ar.iiarjournals.org/ SN - 0250-7005 ER - TY - JOUR A1 - Schlör, Anja A1 - Holzlöhner, Pamela A1 - Listek, Martin A1 - Grieß, Cindy A1 - Butze, Monique A1 - Micheel, Burkhard A1 - Hentschel, Christian A1 - Sowa, Mandy A1 - Roggenbuck, Dirk A1 - Schierack, Peter A1 - Füner, Jonas A1 - Schliebs, Erik A1 - Goihl, Alexander A1 - Reinhold, Dirk A1 - Hanack, Katja T1 - Generation and validation of murine monoclonal and camelid recombinant single domain antibodies specific for human pancreatic glycoprotein 2 JF - New biotechnology N2 - Pancreatic secretory zymogen-granule membrane glycoprotein 2 (GP2) has been identified as a major autoantigenic target in Crohn’s disease patients. It was reported recently that a long (GP2a) and a short (GP2b) isoform of GP2 exist and that in the outcome of inflammatory bowel diseases (IBD) GP2-specific autoantibodies probably appear as new serological markers for diagnosis and therapeutic monitoring. To investigate this further and in order to establish diagnostic tools for the discrimination of both GP2 isoforms, a set of different murine monoclonal and camelid recombinant single domain antibodies (camelid VHH) was generated and validated in various enzyme-linked immunosorbent assay (ELISA) formats, immunofluorescence on transgenic cell lines and immunohistochemistry on monkey pancreas tissue sections. Out of six binders identified, one was validated as highly specific for GP2a. This murine monoclonal antibody (mAb) was used as capture antibody in construction of a sandwich ELISA for the detection of GP2a. Camelid VHHs or a second murine mAb served as detection antibodies in this system. All antibodies were also able to stain GP2a or GP2b on transgenic cell lines as well as on pancreatic tissue in immunohistochemistry. The KD values measured for the camelid VHHs were between 7 nM and 23pM. This set of specific binders will enable the development of suitable diagnostic tools for GP2-related studies in IBD. KW - glycoprotein GP2 KW - Monoclonal antibodies KW - Camelid single domain antibodies Y1 - 2018 U6 - https://doi.org/10.1016/j.nbt.2018.03.006 SN - 1871-6784 SN - 1876-4347 VL - 45 SP - 60 EP - 68 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Schenk, Jörg A. A1 - Sellrie, Frank A1 - Böttger, Volker A1 - Micheel, Burkhard A1 - Stöcklein, Walter F. M. T1 - Generation and application of a fluorescein-specific single chain antibody N2 - A recombinant single chain antibody fragment (designated scDE1) of the murine monoclonal anti-fluorescein antibody B13-DE1 was generated using the original hybridoma cells as source for the variable antibody heavy and light chain (VH and VL) genes. After cloning the variable genes into a phage vector a functional antibody fragment was selected by phage display panning. Recombinant antibody could be expressed as phage antibody and as soluble single chain antibody in Escherichia coli. High yield of scDE1 could also be detected in bacterial culture supernatant. The scDE1 showed the same binding specificity as the parental monoclonal antibody, i.e. it bound fluorescein, fluorescein derivatives and a fluorescein peptide mimotope. Surface plasmon resonance revealed a K(D) of 19 nM for the scDE1 compared to 0.7 nM for the monoclonal antibody. The isolated soluble scDE1 could easily be conjugated to horseradish peroxidase which allowed the use of the conjugate as universal indicator for the detection of fluorescein-labelled proteins in different immunoassays. Detection of hCG in urine was performed as a model system using scDE1. In addition to E. coli the scFv genes could also be transferred and expressed in eukaryotic cells. Finally, we generated HEK293 cells expressing the scDE1 at the cell surface. Y1 - 2007 UR - http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VRJ-4P3DY33- 1&_user=1584062&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000053886&_version=1&_urlVersion=0&_userid=1584062&md5=e 4 ER - TY - JOUR A1 - Gedvilaite, Alma A1 - Frömmel, C. A1 - Sasnauskas, K. A1 - Micheel, Burkhard A1 - Özel, M. A1 - Behrsing, Olaf A1 - Staniulis, J. A1 - Jandrig, Burkhard A1 - Scherneck, Siegfried A1 - Ulrich, R. T1 - Formation of immunogenic virus-like particles by inserting epitopes into surface-exposed regions of hamster polyomavirus major capsid protein Y1 - 2000 ER - TY - JOUR A1 - Hess, Anne-Katrin A1 - Bartel, Manuela A1 - Roth, Karina A1 - Messerschmidt, Katrin A1 - Heilmann, Katja A1 - Kenchington, Ellen A1 - Micheel, Burkhard A1 - Stuckas, Heiko T1 - Expression of M6 and M7 lysin in Mytilus edulis is not restricted to sperm, but occurs also in oocytes and somatic tissue of males and females JF - Molecular reproduction and development N2 - Sperm proteins of marine sessile invertebrates have been extensively studied to understand the molecular basis of reproductive isolation. Apart from molecules such as bindin of sea urchins or lysin of abalone species, the acrosomal protein M7 lysin of Mytilus edulis has been analyzed. M7 lysin was found to be under positive selection, but mechanisms driving the evolution of this protein are not fully understood. To explore functional aspects, this study investigated the protein expression pattern of M7 and M6 lysin in gametes and somatic tissue of male and female M. edulis. The study employs a previously published monoclonal antibody (G26-AG8) to investigate M6 and M7 lysin protein expression, and explores expression of both genes. It is shown that these proteins and their encoding genes are expressed in gametes and somatic tissue of both sexes. This is in contrast to sea urchin bindin and abalone lysin, in which gene expression is strictly limited to males. Although future studies need to clarify the functional importance of both acrosomal proteins in male and female somatic tissue, new insights into the evolution of sperm proteins in marine sessile invertebrates are possible. This is because proteins with male-specific expression (bindin, lysin) might evolve differently than proteins with expression in both sexes (M6/M7 lysin), and the putative function of both proteins in females opens the possibility that the evolution of M6/M7 lysin is under sexual antagonistic selection, for example, mutations beneficial to the acrosomal function that are less beneficial the function in somatic tissue of females.Mol. Reprod. Dev. 79: 517-524, 2012. Y1 - 2012 U6 - https://doi.org/10.1002/mrd.22056 SN - 1040-452X VL - 79 IS - 8 SP - 517 EP - 524 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Kneissel, Sandra A1 - Queitsch, Iris A1 - Petersen, Gabriele A1 - Behrsing, Olaf A1 - Micheel, Burkhard A1 - Düberl, Stefan T1 - Epitope structures recognised by antibodies against the major coat protein (g8p) of filamentous bacteriophage fd (Inoviridae) Y1 - 1999 ER - TY - JOUR A1 - Scheller, Frieder W. A1 - Makower, Alexander A1 - Ghindilis, A. L. A1 - Bier, Frank Fabian A1 - Ehrentreich-Förster, Eva A1 - Wollenberger, Ursula A1 - Bauer, Christian G. A1 - Micheel, Burkhard A1 - Pfeiffer, Dorothea A1 - Szeponik, Jan A1 - Michael, N. A1 - Kaden, H. T1 - Enzyme sensors for subnanomolar concentrations Y1 - 1995 ER - TY - JOUR A1 - Stöcklein, Walter F. M. A1 - Behrsing, Olaf A1 - Scharte, Gudrun A1 - Micheel, Burkhard A1 - Benkert, Alexander A1 - Schössler, W. A1 - Warsinke, Axel A1 - Scheller, Frieder W. T1 - Enzyme kinetic assays with surface plasmon resonance (BIAcore) based on competition between enzyme and creatinine antibody Y1 - 2000 ER - TY - JOUR A1 - Warncke, Max A1 - Vogt, Birgit A1 - Ulrich, Jacqueline A1 - von Laer, Meike Dorothee A1 - Beyer, Winfried A1 - Klump, Hannes A1 - Micheel, Burkhard A1 - Sheriff, Ahmed T1 - Efficient in vitro transduction of naive murine B cells with lentiviral vectors N2 - The aim of this study was to determine the impact of lentiviral transduction on primary murine B cells. Studying B cell activities in vivo or using them for tolerance induction requires that the cells remain unaltered in their biological behavior except for expression of the transgene. As we show here, murine B cells can efficiently be transduced by lentiviral, VSV-G-pseudotyped vectors without the necessity of prior activation. Culture with LPS gave enhanced transduction efficiencies but led to the upregulation of CD86 and proliferation of the cells. Transduction of naive B cells by lentiviral vectors was dependent on multiplicity of infection and did not lead to a concomitant activation. Furthermore, the transduced cells could be used for studies in the NOD mouse system without altering the onset of diabetes. We conclude that lentiviral gene transfer into naive B cells is a powerful tool for manipulation of B cells for therapeutic applications. Y1 - 2004 UR - http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WBK-4C707VR- 7&_coverDate=06%2F04%2F2004&_alid=269000954&_rdoc=1&_fmt=&_orig=search&_qd=1&_cdi=6713&_sort=d&view=c&_acct=C000053886&_v e ER - TY - JOUR A1 - Stöcklein, Walter F. M. A1 - Warsinke, Axel A1 - Micheel, Burkhard A1 - Kempter, Gerhard A1 - Höhne, Wolfgang A1 - Scheller, Frieder W. T1 - Diphenylurea hapten sensing with a monoclonal antibody and its Fab fragment : kinetic and thermodynamic investigations Y1 - 1998 ER - TY - JOUR A1 - Stöcklein, Walter F. M. A1 - Warsinke, Axel A1 - Micheel, Burkhard A1 - Höhne, Wolfgang A1 - Woller, Jochen A1 - Kempter, Gerhard A1 - Scheller, Frieder W. T1 - Detection of diphenylurea derivatives with biospecific interaction analysis (BIA) : Kinetic investigations Y1 - 1997 ER - TY - JOUR A1 - Stuckas, Heiko A1 - Messerschmidt, Katrin A1 - Putzler, Sascha A1 - Baumann, Otto A1 - Schenk, Jörg A. A1 - Tiedemann, Ralph A1 - Micheel, Burkhard T1 - Detection and characterization of gamete-specific molecules in Mytilus edulis using selective antibody production N2 - The mussel Mytilus edulis can be used as model to study the molecular basis of reproductive isolation because this species maintains its species integrity, despite of hybridizing in zones of contact with the closely related species M. trossulus or M. galloprovincialis. This study uses selective antibody production by means of hybridoma technology to identify molecules which are involved in sperm function of M. edulis. Fragmented sperm were injected into mice and 25 hybridoma cell clones were established to obtain monoclonal antibodies (mAb). Five clones were identified producing mAb targeting molecules putatively involved in sperm function based on enzyme immunoassays, dot and Western blotting as well as immunostaining of tissue sections. Specific localization of these mAb targets on sperm and partly also in somatic tissue suggests that all five antibodies bind to different molecules. The targets of the mAb obtained from clone G26-AG8 were identified using mass spectrometry (nano-LC-ESI-MS/MS) as M6 and M7 lysin. These acrosomal proteins have egg vitelline lyses function and are highly similar (76%) which explains the cross reactivity of mAb G26- AG8. Furthermore, M7 lysin was recently shown to be under strong positive selection suggesting a role in interspecific reproductive isolation. This study shows that M6 and M7 lysin are not only found in the sperm acrosome but also in male somatic tissue of the mantle and the posterior adductor muscle, while being completely absent in females. The monoclonal antibody G26-AG8 described here will allow elucidating M7/M6 lysin function in somatic and gonad tissue of adult and developing animals. Y1 - 2009 UR - http://www3.interscience.wiley.com/cgi-bin/jhome/37692 U6 - https://doi.org/10.1002/Mrd.20916 SN - 1040-452X ER - TY - CHAP A1 - Heilmann, Katja A1 - Wand, Inga A1 - Holzlöhner, Pamela A1 - Micheel, Burkhard T1 - Cooperation of dendritic cells with naive lymphocyte populations to induce the generation of antigen-specific antibodies in vitro T2 - The journal of immunology Y1 - 2012 SN - 0022-1767 VL - 188 IS - 6 PB - American Assoc. of Immunologists CY - Bethesda ER - TY - JOUR A1 - Wand, Inga A1 - Holzlöhner, Pamela A1 - Neupert, Steffi A1 - Micheel, Burkhard A1 - Heilmann, Katja T1 - Cooperation of dendritic cells with naive lymphocyte populations to induce the generation of antigen-specific antibodies in vitro JF - Journal of biotechnology N2 - The production of monoclonal antibodies by hybridoma technology is dependent on lymphocytes taken from vertebrates which have to be immunized against the corresponding antigen. We present here our first experiments which should allow the replacement of this in vivo immunization step by an in vitro immunization procedure. This work provides new possibilities for the specific activation of immune cells in order to use them for the generation of antibodies which are not of murine origin. Bone marrow-derived dendritic cells were loaded with antigen and co-cultured with naive T and B lymphocytes of non-immunized mice. The interaction and activation of the different cell types were investigated by measuring the expression of specific cell surface markers, the release of activation-dependent interleukins and the secretion of antigen-specific antibodies. We could demonstrate that dendritic cells process and present antigen fragments and activate T cells, that T cells proliferate and release activation-induced interleukins, and that B cells maturate under the influence of activated T cells and secrete antigen-specific antibodies. KW - In vitro immunization KW - Activation of dendritic cells KW - Interaction of T and B cells with antigen-presenting cells KW - Induction of antibody responses Y1 - 2011 U6 - https://doi.org/10.1016/j.jbiotec.2011.09.002 SN - 0168-1656 VL - 156 IS - 3 SP - 173 EP - 181 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Sellrie, Frank A1 - Schenk, Jörg A. A1 - Behrsing, Olaf A1 - Drechsel, Oliver A1 - Micheel, Burkhard T1 - Cloning and characterization of a single chain antibody to glucose oxidase from a murine hybridoma N2 - Glucose oxidase (GOD) is an oxidoreductase catalyzing the reaction of glucose and oxygen to peroxide and gluconolacton (EC 1.1.3.4.). GOD is a widely used enzyme in biotechnology. Therefore the production of monoclonal antibodies and antibody fragments to GOD are of interest in bioanalytics and even tumor therapy. We describe here the generation of a panel of monoclonal antibodies to native and heat inactivated GOD. One of the hybridomas, E13BC8, was used for cloning of a single chain antibody (scFv). This scFv was expressed in Escherichia coli XL1-blue with the help of the vector system pOPE101. The scFv was isolated from the periplasmic fraction and detected by western blotting. It reacts specifically with soluble active GOD but does not recognize denatured GOD adsorbed to the solid phase. The same binding properties were also found for the monoclonal antibody E13BC8. Y1 - 2007 UR - http://www.jbmb.or.kr/fulltext/jbmb/view.php?vol=40&page=875 ER - TY - JOUR A1 - Lawatscheck, Robert A1 - Aleksaite, Egle A1 - Schenk, Jörg A. A1 - Micheel, Burkhard A1 - Jandrig, Burkhard A1 - Holland, Gudrun A1 - Sasnauskas, Kestutius A1 - Gedvilaite, Alma A1 - Ulrich, Rainer Günter T1 - Chimeric polyomavirus-derived virus-like particles : the immunogenicity of an inserted peptide applied without adjuvant to mice depends on its insertion site and its flanking linker sequence N2 - We inserted the sequence of the carcinoembryonic antigen-derived T cell epitope CAP-1-6D (CEA) into different positions of the hamster polyomavirus major capsid protein VP1. Independently from additional flanking linkers, yeast- expressed VP1 proteins harboring the CEA insertion between VP1 amino acid residues 80 and 89 (site 1) or 288 and 295 (site 4) or simultaneously at both positions assembled to chimeric virus-like particles (VLPs). BALB/c mice immunized with adjuvant-free VLPs developed VP1- and epitope-specific antibodies. The level of the CEA-specific antibody response was determined by the insertion site, the number of inserts, and the flanking linker. The strongest CEA-specific antibody response was observed in mice immunized with VP1 proteins harboring the CEA insert at site 1. Moreover, the CEA- specific antibodies in these mice were still detectable 6 mo after the final booster immunization. Our results indicate that hamster polyomavirus-derived VLPs represent a highly immunogenic carrier for foreign insertions that might be useful for clinical and therapeutic applications. Y1 - 2007 UR - http://www.liebertonline.com/vim U6 - https://doi.org/10.1089/vim.2007.0023 SN - 0882-8245 ER - TY - JOUR A1 - Stöcklein, Walter F. M. A1 - Warsinke, Axel A1 - Micheel, Burkhard A1 - Höhne, Wolfgang A1 - Woller, Jochen A1 - Kempter, Gerhard A1 - Scheller, Frieder W. T1 - Characterization of a monoclonal antibody and its Fab fragment against diphenylurea hapten with BIA Y1 - 1998 SN - 3-8154-3540-4 ER - TY - GEN A1 - Hanack, Katja A1 - Schloer, Anja A1 - Holzloehner, Pamela A1 - Listek, Martin A1 - Bauer, Cindy A1 - Butze, Monique A1 - Micheel, Burkhard A1 - Hentschel, Christian A1 - Sowa, Mandy A1 - Roggenbuck, Dirk A1 - Schierack, Peter A1 - Fuener, Jonas A1 - Schliebs, Erik A1 - Goihl, Alexander A1 - Reinhold, Dirk T1 - Camelid nanobodies specific to human pancreatic glycoprotein 2 T2 - The journal of immunology N2 - Pancreatic secretory zymogen-granule membrane glycoprotein 2 (GP2) has been identified to be a major autoantigenic target in Crohn’s disease patients. It was discussed recently that a long and a short isoform of GP2 exists whereas the short isoform is often detected by GP2-specific autoantibodies. In the outcome of inflammatory bowel diseases, these GP2-specific autoantibodies are discussed as new serological markers for diagnosis and therapeutic monitoring. To investigate this further, camelid nanobodies were generated by phage display and selected against the short isoform of GP2 in order to isolate specific tools for the discrimination of both isoforms. Nanobodies are single domain antibodies derived from camelid heavy chain only antibodies and characterized by a high stability and solubility. The selected candidates were expressed, purified and validated regarding their binding properties in different enzyme-linked immunosorbent assays formats, immunofluorescence, immunohistochemistry and surface plasmon resonance spectroscopy. Four different nanobodies could be selected whereof three recognize the short isoform of GP2 very specifically and one nanobody showed a high binding capacity for both isoforms. The KD values measured for all nanobodies were between 1.3 nM and 2.3 pM indicating highly specific binders suitable for the application as diagnostic tool in inflammatory bowel disease. Y1 - 2016 SN - 0022-1767 SN - 1550-6606 VL - 196 SP - 313 EP - 328 PB - American Assoc. of Immunologists CY - Bethesda ER -