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  • 1
    Keywords: PEPTIDE ; CELLS ; EXPRESSION ; IN-VITRO ; tumor ; TUMOR-CELLS ; Germany ; human ; PROTEIN ; PROTEINS ; TUMORS ; MICE ; RELEASE ; COMPLEX ; RESPONSES ; COMPLEXES ; SERA ; REDUCTION ; DENDRITIC CELLS ; BINDING ; SEQUENCE ; antibodies ; NEUTRALIZING ANTIBODIES ; PARTICLES ; ASSAY ; ESCHERICHIA-COLI ; GLUTATHIONE ; LYMPHOCYTES ; VIRUS-LIKE PARTICLES ; HPV ; HPV16 ; VACCINE ; IMMUNE-RESPONSE ; vaccination ; L1 ; SEQUENCE-ANALYSIS ; glutathione-S-transferase ; HPV PSEUDOVIRIONS ; NASAL IMMUNIZATION
    Abstract: We analyzed capsomeres of human papillomavirus type 16 (HPV16) consisting of the L1 major structural protein for their ability to trigger a cytotoxic T-cell (CTL) response. To this end, we immunized C57BL/6 mice and used the L1(165-173) peptide for ex vivo restimulation of splenocytes prior to analysis (Cr-51 release assay and enzyme-linked immunospot assay [ELISPOT]). This peptide was identified in this study as a D-b-restricted naturally processed CTL epitope by HPV16 L1 sequence analysis, major histocompatibility complex class I binding, and Cr-51 release assays following immunization of C57BL/6 mice with HPV16 L1 virus-like particles (VLPs). HPV16 L1 capsomeres were obtained by purification of HPV16 L1 lacking 10 N-terminal amino acids after expression in Escherichia coli as a glutathione S-transferase fusion protein (GST-HPV16 L1DeltaN10). Sedimentation analysis revealed that the majority of the purified protein consisted of pentameric capsomeres, and assembled particles were not observed in minor contaminating higher-molecular-weight material. Subcutaneous (s.c.) as well as intranasal immunization of C57BL/6 m ice with HPV16 L1 capsomeres triggered an L1-specific CTL response in a dose- dependent manner as measured by ELISPOT and Cr-51 release as say. Significant reduction of contaminating bacterial endotoxin (lipopolysaccharide) from the capsomere preparation did not diminish the immunogenicity. Antibody responses (serum and vaginal) were less robust under the experimental conditions employed. In addition, s.c. vaccination with HPV16 L1 capsomeres induced regression of established tumors expressing L1 determinants (C3 tumor cells). Our data demonstrate that capsomeres are potent inducers of CTL responses similar to completely assembled T=7 VLPs. This result is of potential relevance for the development of (combined prophylactic and therapeutic) HPV-specific vaccines, since capsomeres can be produced easily and also can be modified to incorporate heterologous sequences such as early HPV proteins
    Type of Publication: Journal article published
    PubMed ID: 12663770
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  • 2
    Keywords: IN-VITRO ; Germany ; human ; GENERATION ; PROTEIN ; PROTEINS ; MICE ; RESPONSES ; ANTIGEN ; ANTIGENS ; LYMPH-NODES ; papillomavirus ; antibodies ; antibody ; NEUTRALIZING ANTIBODIES ; PARTICLES ; virus ; CERVICAL-CANCER ; CAPSID PROTEIN ; human papillomavirus ; TYPE-16 ; VIRUS-LIKE PARTICLES ; HPV ; HUMAN-PAPILLOMAVIRUS ; VACCINE ; L1 ; NASAL IMMUNIZATION ; FUSION PROTEIN ; CHOLERA-TOXIN ; CYTOTOXIC T-LYMPHOCYTES ; SERUM ; IMMUNIZATION ; LEVEL ; immunology ; CTA1-D2D1 ; capsomeres ; CTA1-DD VACCINE ADJUVANT ; CTB ; HUMAN PAPILLOMAVIRUSES ; intranasal immunization ; MUCOSAL IMMUNIZATION
    Abstract: Prophylactic immunization against human papillomaviruses (HPVs) aims preferentially at the generation of antibodies, which are directed against the virus capsid proteins. DNA-free virus-like particles or their pentameric subunits, the capsomeres represent suitable antigens. Here we investigated if anti-HPV16 L1 specific antibodies and U-specific CTL induced by intranasal immunization with capsomeres in sera and vaginal washings of C57B16 mice can be enhanced by co-application of the non-toxic cholera toxin adjuvants CTA1-D2D1 or CTB. We found that CTA1-D2D1 elevated L1-specific serum IgG antibodies in a dose-dependent manner and both CTA1-D2D1 and CTB significantly increased L1-specific IgA antibody levels in the vaginal lumen. Furthermore, CTA1-D2D I and CTB enhanced L1-specific CTL responses. (c) 2005 Elsevier Ltd. All rights reserved
    Type of Publication: Journal article published
    PubMed ID: 16455165
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  • 3
    Keywords: CELLS ; EXPRESSION ; IN-VITRO ; CELL ; Germany ; human ; IN-VIVO ; VITRO ; VIVO ; DISEASE ; DISEASES ; PROTEIN ; MOLECULES ; MICE ; ACTIVATION ; LIGAND ; RESPONSES ; DNA ; MARKER ; INDUCTION ; ANTIGEN ; DENDRITIC CELLS ; T-CELLS ; papillomavirus ; MOLECULE ; BONE-MARROW ; MATURATION ; MOUSE ; LESIONS ; HUMANS ; MARKERS ; human papillomavirus ; TYPE-16 ; VACCINES ; PARAMETERS ; HUMAN-PAPILLOMAVIRUS ; MHC CLASS-I ; ONCOPROTEIN ; VACCINE ; vaccination ; T lymphocyte ; FLUORESCENCE ; ANTITUMOR IMMUNITY ; TYPE-2 ; chemokine ; CTL ; TOLL-LIKE RECEPTORS ; CYTOKINE ; ELISA ; RE ; PATTERN ; INCREASE ; TRANSFECTION ; mRNA ; CD40 LIGAND ; methods ; dendritic cell ; pharmacology ; BONE ; ENGLAND ; comparison ; vehicle ; VEHICLES ; MARROW ; CYTOTOXIC ACTIVITY ; CD86 ; CTL RESPONSE ; SOLUBLE CD40 LIGAND
    Abstract: Background: Dendritic cells (DCs) mediate the generation of strong cytotoxic T-lymphocyte (CTL) responses by functioning in antigen presentation and exerting adjuvant properties. We compared several activation markers and parameters of biological activity of DNA- and mRNA-transfected DCs in vitro and in vivo. Methods: CpG-matured, bone marrow derived C57BL/6 mouse DCs were electroporated either with enhanced green fluorescence protein (EGFP) or human papillomavirus type 16 (HPV16) E7 expression plasmids or in vitro transcribed mRNAs encoding for the codon-optimized E7 or a shuffled version thereof. Activation marker expression and antigen presentation was analysed by fluorescence-activated cell sorting. The migratory behaviour of transfected DCs were investigated by in vitro chemotaxis experiments and cytokine expression by ELISA. CTL-priming capacity of transfected DCs were determined by vaccination of mice. Results: mRNA transfection produced a two- to fourfold increase of the activation markers CD40, CD80, CD86 and MHC I and MHC 11 molecules. Predominately antigenexpressing DCs migrated after mRNA transfection. Furthermore, mRNA-transfected DCs were capable of inducing a chemokine gradient. After maturation, electroporation and activation with soluble CD40 ligand and interferon-y, DCs displayed a T-helper cell type 2 cytokine expression pattern. Nevertheless, E7-transfected DCs were able to prime E7-specific CTL responses in vivo. The highest E7-specific CTL frequencies were found in mice immunized with mRNA-transfected DCs. The in vitro expanded CTLs exerted functional E7-specific cytotoxic activity. Conclusions: Genetically modified DCs are suitable vehicles for the induction of U-specific CTL responses in mice and hence could help to eradicate HPV-associated lesions in humans
    Type of Publication: Journal article published
    PubMed ID: 18672528
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  • 4
    Keywords: tumor ; Germany ; human ; TUMORS ; MICE ; RESPONSES ; INDUCTION ; DENDRITIC CELLS ; ESCHERICHIA-COLI ; CERVICAL-CANCER ; antigen presentation ; CHOLERA-TOXIN ; HEAT-LABILE ENTEROTOXIN ; IMMUNIZATION ; PROTEIN-A ; T helper cell ; cytotoxic T lymphocyte ; ADP-RIBOSYLATING EXOTOXINS ; CPG-OLIGONUCLEOTIDES ; CTA1-D2D1 ; human papillomavirus type 16 E7 (HPV-16 E7) ; ovalbumin ; transcutaneous immunization ; tumor protection ; VACCINE ADJUVANT
    Type of Publication: Journal article published
    PubMed ID: 16052529
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  • 5
    ISSN: 1520-5126
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1089-7550
    Source: AIP Digital Archive
    Topics: Physics
    Notes: The magnetic behavior of Ni(SCN)2 has been studied at low temperatures for the first time. A Curie–Weiss fit, χM=C/(T−θ), to the susceptibility between 100 and 300 K yields g=2.13±0.01 (S=1) and θ=39.8±1.4 K. Systematic curvature in χ−1 vs T is evident, however. Despite the large positive θ Ni(SCN)2 appears to order antiferromagnetically at Tc=52±1 K, slightly below a maximum in χ(T) at T(χmax)=57.2±0.5 K, with χmax=0.0331±0.0003 emu/mol. The ratio Tc/T(χmax)=0.91±0.02 does not suggest lower magnetic dimensionality. Magnetization isotherms are linear to 16 kG; some features suggesting lower temperature transitions occur. Well above Tc the susceptibility is analyzed assuming axial and rhombic crystal field distortions, i.e., D[Sˆz2−S(S+1)/3] and E[Sˆx2−Sˆy2] spin Hamiltonian terms, with exchange incorporated in a mean field approximation. An extraordinarily large ||D/k||≈119 K seems to emerge, a result which is very provisional lacking single crystal data. A mean field analysis of Tc and θ yields ferromagnetic intrachain exchange J1/k=8.6±0.5 K and antiferromagnetic interchain exchange J2/k=−0.76±0.4 K. It seems more likely that D is negative, but even if it is positive the exchange interaction is large enough to induce magnetic order at finite Tc in light of theories relating Tc, J, and D.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1089-7550
    Source: AIP Digital Archive
    Topics: Physics
    Notes: Previous magnetization and susceptibility measurements on the mixed magnet Co1−xMnxCl2⋅2H2O disclosed a spin glass transition near 2.45 K over a wide composition range.1 Recent heat capacity and NMR measurements have confirmed and extended this finding.2 The time dependence of the thermoremanent magnetization (TRM) below Tg for an x=0.452 mixture was studied in some detail1 and was found to conform approximately to a decay of stretched exponential type, or perhaps slightly more accurately to the product of a stretched exponential and a power law. Small systematic deviations of data from fitted curves were apparent however. Recently a percolation model for relaxation in random systems was proposed,3 and yielded significantly improved fits to the TRM decay in an Au:Fe spin glass. The model assumes dispersive excitations within fixed finite domains, and includes as parameters the fastest and slowest relaxation rates characterizing the spectrum of domains. We find that this model also permits much better fits to be obtained for the TRM decay in Co1−xMnxCl2⋅2H2O, x=0.452. Systematic deviations that were present when using more traditional decay functions are virtually eliminated. The variation of fitted parameters with cooling field and temperature is also explored.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1089-7550
    Source: AIP Digital Archive
    Topics: Physics
    Notes: The magnetic properties of Ni(SCN)2(C2H5OH)2 have been studied, the first Ni(II) system in the general family of compounds M(SCN)2(ROH)2 (where M=divalent Mn, Fe, Co, or Ni and R=CH3, C2H5, i- or n-C3H7) to be examined at low temperatures. In contrast to previously studied Mn(II) and Co(II) members of this family, which exhibit predominant antiferromagnetism, the present compound is ferromagnetic. The susceptibility of a polycrystalline sample is of Curie–Weiss form only above 75 K, with g¯ = 2.175 ± 0.01 and S=1 and with θ=24.1±1.0 K. The initial susceptibility is well accounted for by an asymptotic critical law, χ0 = Γ[T/Tc − 1]−γ, in the reduced temperature range 0.147–0.013, with Tc = 13.081 ± 0.01 K, γ=1.354±0.02, and Γ=0.0925±0.003 emu/mol. The γ value is between 3D-XY and 3D-Heisenberg model predictions. The susceptibility in the paramagnetic regime well above Tc is analyzed including the effects of axial and rhombic crystal field distortions, represented by D[Sˆ2z − S(S + 1)/3] and E[Sˆ2x − Sˆ2y] terms in the spin Hamiltonian, and incorporating exchange interactions in a mean-field approximation. An excellent fit is obtained with g=2.175, D/k=−64.3±5 K, E/k=23.1±4 K, and zJ/k=19.4±1 K. The magnitude of D is the largest we know of in a Ni(II) compound. The three spin states of the 3A crystal field ground term are strongly split, probably contributing to the relatively low value of Tc compared to zJ/k. Some two-dimensional character in the exchange, as has been found for other members of this family of compounds, may also contribute.
    Type of Medium: Electronic Resource
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