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  • 1
    Keywords: CANCER ; EXPRESSION ; tumor ; carcinoma ; Germany ; human ; DISEASE ; GENE ; MONOCLONAL-ANTIBODY ; PATIENT ; DONOR ; ANTIGEN ; SKIN ; BREAST-CANCER ; IMMUNE-RESPONSES ; antibodies ; CELL-LINE ; LINE ; TUMOR-ASSOCIATED ANTIGENS ; CYTOLYTIC T-LYMPHOCYTES ; SERUM ; CELL CARCINOMA ; renal cell carcinoma ; RE ; HUMAN-MELANOMA ; SEREX ; human renal cell carcinoma ; AUTOLOGOUS ANTIBODY ; cancer-testis antigen ; CDNA CLONING ; immunome ; SADA ; VALOSIN-CONTAINING PROTEIN
    Abstract: Serological analysis of cDNA expression libraries (SEREX) has proven to be a useful technique in the quest to elucidate the repertoire of immunogenic gene products in human cancer. We have applied the SEREX method to human renal cell carcinoma (RCC) in order to identify associated immunogenic gene products. cDNA expression libraries were prepared from a RCC tumor, a RCC cell line and human testis. The 3 libraries were screened with sera from 35 RCC patients and 15 healthy controls. Approximately 4.5 x 10(6) phage plaques were screened resulting in 234 positive clones, which corresponded to 74 different gene products. The seroreactivity toward 49 of these antigens was assessed. Seroreactivity to 21 (43%) of the antigens was similar in RCC patients and healthy controls, 9 antigens (18%) elicited antibodies more frequently and 19 antigens (39%) solely in RCC patients. In the reverse setting, reactivity of RCC patients' sera was tested against a panel of 44 previously identified "tumor-associated" antigens via the SADA (serum antibody detection array) method; 6 antigens reacted with RCC patients' and healthy donors' sera, 8 were reactive only with RCC patients' sera. From the 27 antigens identified by SEREX and SADA, which did not react with sera from healthy controls, 10 antigens reacted with a significant proportion of RCC patients' sera and 77% of RCC patients' sera reacted at least with one of these antigens. Sera from patients with nonmalignant renal diseases or an autoimmune disease did not react with these 10 antigens. (c) 2005 Wiley-Liss, Inc
    Type of Publication: Journal article published
    PubMed ID: 16331622
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  • 2
    Abstract: The yeast Saccharomyces cerevisiae is ideal for systematic studies relying on collections of modified strains (libraries). Despite the significance of yeast libraries and the immense variety of available tags and regulatory elements, only a few such libraries exist, as their construction is extremely expensive and laborious. To overcome these limitations, we developed a SWAp-Tag (SWAT) method that enables one parental library to be modified easily and efficiently to give rise to an endless variety of libraries of choice. To showcase the versatility of the SWAT approach, we constructed and investigated a library of approximately 1,800 strains carrying SWAT-GFP modules at the amino termini of endomembrane proteins and then used it to create two new libraries (mCherry and seamless GFP). Our work demonstrates how the SWAT method allows fast and effortless creation of yeast libraries, opening the door to new ways of systematically studying cell biology.
    Type of Publication: Journal article published
    PubMed ID: 26928762
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  • 3
    Keywords: CANCER ; CELLS ; EXPRESSION ; tumor ; carcinoma ; CELL ; Germany ; human ; GENE ; PROTEIN ; PROTEINS ; cell line ; TISSUE ; TUMORS ; LINES ; PATIENT ; MESSENGER-RNA EXPRESSION ; FAMILY ; TISSUES ; ANTIGEN ; ANTIGENS ; SKIN ; T-CELLS ; CELL-LINES ; antibodies ; antibody ; TARGET ; FOCI ; CELL-LINE ; LINE ; MELANOMA ; Jun ; IMMUNOTHERAPY ; MALIGNANT-MELANOMA ; CANCER-IMMUNOTHERAPY ; CANCER/TESTIS ANTIGENS ; MELANOMA PATIENTS ; TARGETS ; malignant melanoma ; cell lines ; AUTOANTIBODIES ; protein expression ; TUMOR-ASSOCIATED ANTIGENS ; TRICHOSTATIN-A ; RABBIT ; ONCOLOGY ; RECOMBINANT ; RE ; FAMILIES ; HUMAN-MELANOMA ; GAGE ; HETEROGENEOUS EXPRESSION ; human melanoma ; analysis ; PHASE-I TRIAL ; trichostatin A ; autoantibody ; IMMUNOHISTOCHEMICAL ANALYSIS ; UNIT ; 5-AZA-2'-DEOXYCYTIDINE ; 5'-aza-2'-deoxycytidine ; serum autoantibodies
    Abstract: Cancer/testis antigens are considered as promising targets for immunotherapy against tumors including malignant melanoma. One group of these antigens is the GAGE antigen family. In this study, we obtained recombinant GAGE-7b protein against which a rabbit antiserum was generated. The polyclonal, monospecific antibodies were used to analyze the expression of GAGE family proteins in human melanoma tissues and cell lines. GAGE expression in melanoma cell lines ranged from 41% to 58% and in melanoma tissues from 22% to 53%. Immunohistochemical analysis of melanoma tumors revealed a rather heterogeneous expression of GAGE resulting in individual positive cells or foci of stained cells. Furthermore, we could show that autoantibodies against GAGE family proteins are detectable in 6% of melanoma patients. Besides, we first demonstrated that the expression of GAGE family proteins can be stimulated with 5'-aza-2'-deoxycytidine and trichostatin A. Through upregulation of protein expression GAGE family proteins might develop into promising targets for immunotherapy of melanoma and other tumors. (C) 2006 Elsevier Ireland Ltd. All rights reserved
    Type of Publication: Journal article published
    PubMed ID: 17194529
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  • 4
    Keywords: CANCER ; CELLS ; EXPRESSION ; tumor ; BLOOD ; CELL ; Germany ; DIAGNOSIS ; GENE ; GENES ; PROTEINS ; TISSUE ; PATIENT ; TISSUES ; ANTIGEN ; SKIN ; T cell ; T-CELL ; SUPPRESSION ; BREAST-CANCER ; antibodies ; IDENTIFICATION ; SSH ; MIGRATION ; HUMAN HOMOLOG ; STRATEGIES ; IMMUNE-RESPONSE ; IMMUNOTHERAPY ; CTCL ; PERIPHERAL-BLOOD ; TUMOR-ASSOCIATED ANTIGENS ; NON-HODGKINS-LYMPHOMA ; MYCOSIS-FUNGOIDES ; SERUM ; RECOMBINANT ; RE ; PATTERN ; ARRAY ; T-CELL LYMPHOMA ; CUTANEOUS LYMPHOMAS ; humoral immune response ; SEREX ; analysis ; TESTS ; ENGLAND ; SADA ; SEROLOGICAL ANALYSIS
    Abstract: The knowledge of tumor-associated antigens is required for most types of immunotherapy and can substantially facilitate diagnosis. To identify potential tumor-associated genes expressed in cutaneous T-cell lymphoma (CTCL), we used three complementary strategies: antigens which elicit a humoral immune response in CTCL patients were detected by serological analysis of a recombinant cDNA expression library. cDNAs differentially expressed in CTCL but not peripheral blood monocytes were identified by comparative cDNA hybridization and suppression subtractive hybridization. We identified 43 genes selectively expressed by CTCL cells, that have not yet been described in the context of CTCL development, but most of which had been reported to be associated with cancer. Expression analysis by database mining and subsequently RT-PCR on selected clones confirmed their selective expression in CTCL tissues. Serological tests showed that 15 clones were recognized by sera of CTCL patients but not of healthy donors. Analysis of serological tests for 11 clones using serum antibody detection array (SADA) and 100 sera of controls and CTCL patients each revealed up to 5% reactive sera in the tumor group. The expression pattern of the detected clones and their immunogenicity demonstrates that they might be relevant for the understanding of CTCL and suggests particularly three clones, HD-CL-41 (DRAK2), HD-CL-49 (nudC) and HD-CL-12 (ZNF195) for further analysis with respect to their prognostic and therapeutic value for CTCL
    Type of Publication: Journal article published
    PubMed ID: 17979976
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  • 5
    Keywords: tumor ; BLOOD ; IN-VIVO ; THERAPY ; CLASSIFICATION ; SITE ; PATIENT ; ACTIVATION ; INDUCTION ; ANTIGEN ; ANTIGENS ; T-CELL ; ASSOCIATION ; antibodies ; antibody ; TARGET ; TRIAL ; LESIONS ; LYMPHOMA ; genetics ; IMMUNOTHERAPY ; TARGETS ; PERIPHERAL-BLOOD ; B-CELL LYMPHOMA ; PHASE-II ; MYCOSIS-FUNGOIDES ; DISORDERS ; REGRESSION ; monitoring ; SEZARY-SYNDROME ; PHASE ; LYMPHOMAS ; tumor antigen ; B-CELL LYMPHOMAS ; prospective ; RECOMMENDATIONS ; CLINICAL-RESPONSE ; B-CELL ; EORTC ; Genetic ; IMMUNE ; PROPOSAL ; Type ; CLINICAL-RESPONSES ; Cutaneous ; LYMPHOPROLIFERATIVE DISORDER ; TASK-FORCE ; TREATMENT-OF-CANCER
    Abstract: Cutaneous lymphomas (CLs) are a heterogeneous group of lymphoproliferative disorders that are manageable by immunotherapy. Twenty-one patients were enrolled in a prospective open-label, dose-escalation multi-center study evaluating the effects of repeated TG1042 [adenovirus-interferon (IFN)-gamma] intralesional injections in patients with primary CLs, of which 18 were of T-cell and 3 of B-cell type. Repeated intralesional therapy using TG1042 consistently results in local tumor regressions in about half of treated patients and one-third of patients also in regressions in noninjected distant lesions, likely reflecting the systemic immune activation after intralesional therapy. Treatment was well tolerated with few adverse events including injection site reactions, chills, lymphopenia, and fever. Immune monitoring in the peripheral blood demonstrated systemic immune activation and the induction of antibodies against tumor antigens in some patients without clear association with clinical responses. CLs, in particular B-cell lymphomas with high objective response rates, seem to be excellent targets for this type of immunotherapy
    Type of Publication: Journal article published
    PubMed ID: 20372104
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  • 6
    Publication Date: 2018-01-19
    Description: The biogenesis of mitochondria, chloroplasts, and Gram-negative bacteria requires the insertion of β-barrel proteins into the outer membranes. Homologous Omp85 proteins are essential for membrane insertion of β-barrel precursors. It is unknown if precursors are threaded through the Omp85-channel interior and exit laterally or if they are translocated into the membrane at the Omp85-lipid interface. We have mapped the interaction of a precursor in transit with the mitochondrial Omp85-channel Sam50 in the native membrane environment. The precursor is translocated into the channel interior, interacts with an internal loop, and inserts into the lateral gate by β-signal exchange. Transport through the Omp85-channel interior followed by release through the lateral gate into the lipid phase may represent a basic mechanism for membrane insertion of β-barrel proteins.
    Keywords: Cell Biology
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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