Parenteral administration of foreign proteins results in earthworms in significantly increased levels of the antigen-binding protein (ABP). The earthworms (Lumbricus terrestris and Eisenia foetida; Oligochaeta, Annelida) were stimulated with different proteins and the ABP response was followed with an anti-ABP monoclonal antibody and in ligand assay with biotin conjugates of all proteins used for stimulation. ABP levels are increased after the stimulation with different proteins irrespective of the size and extent of glycosylation. Molar mass of the ABP molecules was always the same, 56 kDa in L. terrestris and 60 kDa in E. foetida. The level of the specificity is considerably lower in comparison with immunoglobulins since ABP reacts not only with the protein used for stimulation but also, though to lesser extent, with related proteins.

• MeSH
• antigeny heterofilní MeSH
• antigeny * MeSH
• monoklonální protilátky MeSH
• Oligochaeta imunologie   MeSH
• proteiny imunologie   MeSH
• vazba proteinů imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny heterofilní MeSH
• antigeny * MeSH
• monoklonální protilátky MeSH
• proteiny MeSH

Earthworms respond to parenteral stimulation with a protein antigen by the formation of an antigen-binding protein. Earthworms were parenterally stimulated with different proteins and the protein binding was estimated in vitro on both humoral and cellular levels. The binding was significantly higher when the same protein was used for in vivo stimulation. The degree of specificity of the antigen-binding protein after the secondary in vivo challenge increased, but even so it was considerably lower than that of vertebrate immunoglobulins.

• MeSH
• adjuvancia imunologická farmakologie   MeSH
• epitopy MeSH
• hapteny farmakologie   MeSH
• imunizace MeSH
• kyselina arsanilová farmakologie   MeSH
• Oligochaeta imunologie   MeSH
• proteiny imunologie   MeSH
• sérový albumin farmakologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• adjuvancia imunologická MeSH
• epitopy MeSH
• hapteny MeSH
• kyselina arsanilová MeSH
• proteiny MeSH
• sérový albumin MeSH

Activated CD8+ T cells (TCD8+) can directly recognize malignant cells because processed fragments of tumour associated antigens (TAA), 8-10 amino acids in length and complexed with MHC class I molecules, are displayed on tumour cell surfaces. Tumour cells have been genetically modified in a variety of ways in efforts to enhance the immune recognition of TAA. An alternative strategy is the expression of TAA in recombinant or synthetic form. This has been made possible by the recent cloning of TAA recognized by TCD8+. In this communication we review recent work in our own laboratory on the expression of TAA as synthetic peptide, by "naked" plasmid DNA injected intramuscularly or transdermally, and by recombinant viruses including vaccinia (rVV), fowlpox (rFV) and adenovirus (rAd). The expression of TAA in recombinant and synthetic forms allows increased control over the quantity, location, and kinetics of TAA presentation and can result in powerful, specific, anti-tumour immune responses.

• MeSH
• antigeny nádorové genetika   imunologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• imunoterapie MeSH
• interferon gama genetika   terapeutické užití   MeSH
• molekulární sekvence - údaje MeSH
• myši MeSH
• oligopeptidy chemická syntéza   genetika   imunologie   MeSH
• prezentace antigenu * MeSH
• regulace exprese virových genů MeSH
• rekombinantní proteiny imunologie   MeSH
• sekvence aminokyselin MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• antigeny nádorové MeSH
• interferon gama MeSH
• oligopeptidy MeSH
• rekombinantní proteiny MeSH

Prostate-specific membrane antigen (PSMA) is a well-characterized tumor marker associated with prostate cancer and neovasculature of most solid tumors. PSMA-specific ligands are thus being developed to deliver imaging or therapeutic agents to cancer cells. Here, we report on a crystal structure of human PSMA in complex with A9g, a 43-bp PSMA-specific RNA aptamer, that was determined to the 2.2 Å resolution limit. The analysis of the PSMA/aptamer interface allows for identification of key interactions critical for nanomolar binding affinity and high selectivity of A9g for human PSMA. Combined with in silico modeling, site-directed mutagenesis, inhibition experiments and cell-based assays, the structure also provides an insight into structural changes of the aptamer and PSMA upon complex formation, mechanistic explanation for inhibition of the PSMA enzymatic activity by A9g as well as its ligand-selective competition with small molecules targeting the internal pocket of the enzyme. Additionally, comparison with published protein-RNA aptamer structures pointed toward more general features governing protein-aptamer interactions. Finally, our findings can be exploited for the structure-assisted design of future A9g-based derivatives with improved binding and stability characteristics.

• MeSH
• antigeny povrchové chemie   MeSH
• aptamery nukleotidové chemie   MeSH
• buňky PC-3 MeSH
• glutamátkarboxypeptidasa II chemie   MeSH
• HEK293 buňky MeSH
• interakční proteinové domény a motivy MeSH
• lidé MeSH
• ligandy MeSH
• molekulární struktura MeSH
• nádorové biomarkery chemie   MeSH
• nádory prostaty metabolismus   MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• A9g RNA aptamer MeSH Prohlížeč
• antigeny povrchové MeSH
• aptamery nukleotidové MeSH
• FOLH1 protein, human MeSH Prohlížeč
• glutamátkarboxypeptidasa II MeSH
• ligandy MeSH
• nádorové biomarkery MeSH

Interaction between an antigen-presenting cell and a T cell, and their subsequent conjugation are a prerequisite for the formation of the immunological synapse and productive, antigen-dependent activation of T cells. This initial interaction is accompanied by recognition of the presented antigen by the T cell receptor, and by changes in the morphology of the interacting cells and in actin cytoskeleton structure in the site of interaction. The experimental protocol below describes a simple assay for quantitative assessment of antigen-presenting cells-T cell conjugation using confocal microscopy or flow cytometry.

• Klíčová slova
• B3Z, Confocal microscopy, Conjugation assay, Flow cytometry, Immunological synapse, MutuDC,
• MeSH
• aktivace lymfocytů MeSH
• antigen prezentující buňky * MeSH
• imunologické synapse fyziologie   MeSH
• mikrofilamenta MeSH
• receptory antigenů T-buněk MeSH
• T-lymfocyty * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• receptory antigenů T-buněk MeSH

Neonatal transplantation tolerance was induced in B10.A mice by the injection of spleen and bone marrow cells from semiallogeneic [C57BL/10(B10) x B10.A] F1 donors. The neonatally treated mice accepted skin grafts from B10 donors. Spleen cells from tolerant animals did not respond by proliferation to tolerated B10 antigens in vitro. However, spleen cells from tolerant mice recognized specific (B10) antigens and synthesized mRNA for the inducible 55-kDa interleukin-2 receptor (IL-2R) as did cells from normal animals. Maintenance of this early phase of cell activation upon contact with tolerated antigens is direct evidence against clonal deletion as a mechanism, in this particular model of neonatally induced transplantation tolerance.

• MeSH
• biologické modely MeSH
• buňky kostní dřeně MeSH
• exprese genu MeSH
• histokompatibilní antigeny imunologie   MeSH
• imunologická tolerance genetika   MeSH
• messenger RNA biosyntéza   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• novorozená zvířata MeSH
• receptory interleukinu-2 genetika   MeSH
• slezina cytologie   MeSH
• transplantační imunologie genetika   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• histokompatibilní antigeny MeSH
• messenger RNA MeSH
• receptory interleukinu-2 MeSH

Peripheral blood leucocytes from normal, non-immunized adult rats recognize syngeneic and allogeneic tissue antigens, and this recognition can be detected by the tube leucocyte adherence inhibition assay. We show that PBL from newborn rats have the same and even a stronger ability for recognition and that the recognition is preserved in rats bearing tolerated skin allografts after neonatal induction of transplantation tolerance. Testing of tissue extracts prepared from the congenic strain of rats suggested that PBL from normal animals recognize histocompatibility antigens.

• MeSH
• buněčná imunita MeSH
• epitopy MeSH
• histokompatibilní antigeny imunologie   MeSH
• imunizace MeSH
• imunologická tolerance MeSH
• inbrední kmeny potkanů MeSH
• krysa rodu Rattus MeSH
• leukocyty imunologie   MeSH
• novorozená zvířata imunologie   MeSH
• přirozená imunita MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• epitopy MeSH
• histokompatibilní antigeny MeSH

Chemical immunotherapeutic strategies including Antibody Recruiting Molecules (ARMs - bivalent small molecules containing an antibody-binding domain (ABD) and a target-binding domain (TBD)) direct immune-mediated clearance of diseased cells. Anti-cancer ARM function relies on high tumor antigen valency, limiting function against lower antigen expressing tumors. To address this limitation, we report a tunable multivalent immune recruitment (MIR) platform to amplify/stabilize antibody recruitment to cells with lower antigen valencies. An initial set of polymeric ARMs (pARMs) were synthesized and screened to evaluate ABD/TBD copy number, ratio, and steric occlusion on specific immune induction. Most pARMs demonstrated simultaneous high avidity binding to anti-dinitrophenyl antibodies and prostate-specific membrane antigens on prostate cancer. Optimized pARMs mediated enhanced anti-cancer immune function against lower antigen expressing target cells compared to an analogous ARM.

• Klíčová slova
• Antibodies, Cell Recognition, Drug Design, Immunochemistry, Polymers,
• MeSH
• antigeny * MeSH
• fagocytóza MeSH
• lidé MeSH
• nádory prostaty * MeSH
• protilátky chemie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny * MeSH
• protilátky MeSH

Human antigen R (HuR) is a key regulator of cellular mRNAs containing adenylate/uridylate-rich elements (AU-rich elements; AREs). These are a major class of cis elements within 3' untranslated regions, targeting these mRNAs for rapid degradation. HuR contains three RNA recognition motifs (RRMs): a tandem RRM1 and 2, followed by a flexible linker and a C-terminal RRM3. While RRM1 and 2 are structurally characterized, little is known about RRM3. Here we present a 1.9-Å-resolution crystal structure of RRM3 bound to different ARE motifs. This structure together with biophysical methods and cell-culture assays revealed the mechanism of RRM3 ARE recognition and dimerization. While multiple RNA motifs can be bound, recognition of the canonical AUUUA pentameric motif is possible by binding to two registers. Additionally, RRM3 forms homodimers to increase its RNA binding affinity. Finally, although HuR stabilizes ARE-containing RNAs, we found that RRM3 counteracts this effect, as shown in a cell-based ARE reporter assay and by qPCR with native HuR mRNA targets containing multiple AUUUA motifs, possibly by competing with RRM12.

• Klíčová slova
• NMR spectroscopy, RNA-binding protein, crystal structure, dimerization, multiple register,
• MeSH
• 3' nepřekládaná oblast MeSH
• dimerizace MeSH
• encefalomyelitida paraneoplastická - Hu antigeny chemie   MeSH
• HuR protein chemie   genetika   MeSH
• krystalografie rentgenová MeSH
• lidé MeSH
• magnetická rezonanční spektroskopie MeSH
• motiv rozpoznávající RNA genetika   MeSH
• nádorové supresorové proteiny chemie   MeSH
• proteiny vázající RNA chemie   genetika   MeSH
• ribonukleosiddifosfátreduktasa chemie   MeSH
• úseky bohaté na AU genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 3' nepřekládaná oblast MeSH
• encefalomyelitida paraneoplastická - Hu antigeny MeSH
• HuR protein MeSH
• nádorové supresorové proteiny MeSH
• proteiny vázající RNA MeSH
• ribonucleotide reductase M2 MeSH Prohlížeč
• ribonukleosiddifosfátreduktasa MeSH
• RRM1 protein, human MeSH Prohlížeč

Adherence to glass of peripheral blood leucocytes from normal, non-immunized rats was measured in the tube LAI assay in the presence of extracts isolated from tissues of syngeneic or allogeneic donors. Adherence inhibition was significantly higher when peripheral blood leucocytes were incubated with syngeneic than with allogeneic tissue extracts. The results indicate that peripheral blood leucocytes from normal, non-immunized animals are capable of recognizing self and non-self tissue antigens and that this recognition can be detected by the tube LAI assay.

• MeSH
• druhová specificita MeSH
• histokompatibilní antigeny genetika   MeSH
• hlavní histokompatibilní komplex * MeSH
• inbrední kmeny potkanů MeSH
• krysa rodu Rattus MeSH
• leukocyty imunologie   MeSH
• potkani inbrední LEW MeSH
• test inhibice adherence leukocytů MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• histokompatibilní antigeny MeSH