ABSTRACT
A balanced immune response is a cornerstone of healthy aging. Here, we uncover distinctive features of the long-lived blind mole-rat (Spalax spp.) adaptive immune system, relative to humans and mice. The T-cell repertoire remains diverse throughout the Spalax lifespan, suggesting a paucity of large long-lived clones of effector-memory T cells. Expression of master transcription factors of T-cell differentiation, as well as checkpoint and cytotoxicity genes, remains low as Spalax ages. The thymus shrinks as in mice and humans, while interleukin-7 and interleukin-7 receptor expression remains high, potentially reflecting the sustained homeostasis of naive T cells. With aging, immunoglobulin hypermutation level does not increase and the immunoglobulin-M repertoire remains diverse, suggesting shorter B-cell memory and sustained homeostasis of innate-like B cells. The Spalax adaptive immune system thus appears biased towards sustained functional and receptor diversity over specialized, long-lived effector-memory clones-a unique organizational strategy that potentially underlies this animal's extraordinary longevity and healthy aging.
Subject(s)
Spalax , Humans , Mice , Animals , Spalax/genetics , Interleukin-7/metabolism , Mole Rats , Adaptive Immunity , Immunoglobulins/metabolismABSTRACT
Chimeric gp130 receptors were produced to study the role of three fibronectin type III-like domains in activation of gp130 receptor machinery. The ligand-induced dimerization of gp130 was sufficient to trigger STAT3 signaling pathway. These findings can be used as the basis in designing novel therapeutic gp130 inhibitors.
Subject(s)
Cytokine Receptor gp130/metabolism , Fibronectins/pharmacology , Cytokine Receptor gp130/chemistry , Fibronectins/chemistry , HEK293 Cells , Humans , Protein Multimerization/drug effects , STAT3 Transcription Factor/metabolism , Signal Transduction/drug effectsABSTRACT
Multiple Sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system that primarily affects young and middle-aged people. It is widely accepted that B lymphocyte activation is required for MS progression. Despite the fact that the exact triggering mechanisms of MS remain enigmatic, one may suggest that MS can be induced by viral or bacterial infection in combination with specific genetic and environmental factors. Using deep sequencing and functional selection methodologies we characterized clones of poly- and cross-reactive antibodies that are capable of simultaneous recognition of viral proteins and autoantigens. The latter, in turn, possibly may trigger MS progression through molecular mimicry. It was identified that two cross-reactive antigens are probably recognized by light or heavy chains individually. According to the high structural homology between selected autoantibodies and a number of various antiviral IgGs, we suggest that a wide range of pathogens, instead of a single virus, be regarded as possible triggers of MS.
Subject(s)
Drug Resistance, Multiple/genetics , HIV Protease Inhibitors/pharmacology , HIV Protease/genetics , Indinavir/pharmacology , Molecular Dynamics Simulation , Mutation/genetics , Sulfonamides/pharmacology , Amino Acid Substitution/genetics , Darunavir , Drug Design , Fluorescent Dyes/pharmacology , HIV Protease/chemistry , HIV Protease/metabolism , Humans , KineticsABSTRACT
Expression of recombinant antibodies in mammalian cells is one of key problems in immunobiotechnology. Alternatively, expression of a broad panel of antibodies and of their fragments may be effectively done in yeast cells. We obtained expression strains of the methylotrophic beast Pichia pastoris producing single chain human catalytic antibody A17 (A.17scFv), Fab-fragment (A.17Fab) and full-size light chain (A.17Lch). These antibodies were characterized in terms of functional activity. The capacity to specifically bind and transform organophosphorus compounds has been demonstrated for A.17scFv and A.17Fab. The loss of activity of the antibody light chain when expressed alone indicates that the active site is formed by both heavy and light chains of the antibody. We determined the reversible constant Kd and the first order constant (k2) of the reaction of the covalent modification of A.17scFv and A.17Fab by irreversible inhibitor of the serine proteases p-nitrophenyl 8-methyl-8-azobicyclo[3.2.1]phosphonate (Phosphonate X). Calculated values indicate that activity of the antibodies expressed in yeast is similar to the full-size antibody A17 and single chain antibody A.17 expressed in CHO and E. coli cells respectively.
Subject(s)
Antibodies, Catalytic/biosynthesis , Antibodies, Monoclonal/biosynthesis , Gene Expression , Pichia , Recombinant Proteins/biosynthesis , Single-Chain Antibodies/biosynthesis , Animals , Antibodies, Catalytic/genetics , Antibodies, Monoclonal/genetics , CHO Cells , Cricetinae , Cricetulus , Humans , Recombinant Proteins/genetics , Single-Chain Antibodies/geneticsABSTRACT
Proteins of 65 and 57 kD were isolated from the apical membranes of midgut epithelium of Anopheles stephensi larvae by affinity chromatography. These proteins can specifically bind endotoxin Cry11A and activate toxin Cry4B (Cry4B-tox) under conditions of ligand blotting, and both Cry proteins compete for this binding. At least in the case of Cry4B-tox, the binding with 65 and 57 kD proteins is reversible. The ability of the products of limited proteolysis of Cry11A and Cry4B to bind the 65 and 57 kD proteins correlates with their toxicity to A. stephensi larva. The N-terminal amino acid sequence of the 57 kD protein is unique and absent in the NCBI GenBank. The proteins of 65 and 57 kD share most of the properties studied with Aedes aegypti toxin-binding proteins. It is possible that they altogether represent a novel class (or classes) of delta-endotoxin receptors.
Subject(s)
Anopheles/metabolism , Bacterial Proteins/metabolism , Bacterial Toxins/metabolism , Carrier Proteins/metabolism , Endotoxins/metabolism , Intestinal Mucosa/metabolism , Animals , Anopheles/microbiology , Bacillus thuringiensis/metabolism , Bacillus thuringiensis/pathogenicity , Bacillus thuringiensis Toxins , Bacterial Proteins/chemistry , Bacterial Toxins/chemistry , Carrier Proteins/chemistry , Chromatography, Affinity , Endotoxins/chemistry , Hemolysin Proteins , Intestinal Mucosa/enzymology , Intestinal Mucosa/microbiology , Intestinal Mucosa/ultrastructure , Larva/chemistry , Larva/metabolism , Larva/microbiology , Larva/ultrastructure , Microvilli/chemistry , Microvilli/metabolism , Microvilli/microbiology , Microvilli/ultrastructure , Molecular Weight , Phosphoric Monoester Hydrolases/metabolismABSTRACT
The dynamics of pathological changes in the intestine of Aedes aegypti larvae under the influence of toxins Cry11A and Cry4B produced by Bacillus thuringiensis israelensis was studied by means of electron microscope. Most significant ultrastructure changes in the intestine of the second instar larvae were observed in the midgut. The cytoplasm of cells disintegrated, and elongated lacunae appeared. The number of microvilli decreased, or they disappeared in the result of destruction. The peritrophic membrane displaced to the lumen of midgut. Any changes in epithelial cells and cuticle in time of foregut and hindgut were not observed in a comparison to control. The toxin Cry4B caused the most effective destruction of the midgut epithelium.