Active immunity against the CD4 receptor by using an antibody antigenized with residues 41-55 of the first extracellular domain.

Lanza, Paola; Billetta, Rosario; Антоненко, Светлана; Zanetti, Maurizio

doi:10.1073/pnas.90.24.11683

Cited by 17 publications

(7 citation statements)

References 48 publications

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“…The approach of using a protein of known three-dimensional structure to present active sequences in a constrained and structurally defined conformation has already been addressed by other groups. However, in all published examples much larger proteins have been used; immunoglobulin (150 kDa) (37)(38)(39)(40), alkaline phosphatase (121 kDa) (41), interleukin 1,3 (17 kDa) (40,42,43), staphylococcal nuclease (17 kDa) (44), lysozyme (15 kDa) (45,46), and the exogenous sequences have been invariably inserted on exposed loop regions. A newly designed shorter version of the immunoglobulin fold, the minibody (7 kDa) (27), has also been designed and again the loop region was engineered.…”

Section: Methodsmentioning

confidence: 99%

Scorpion toxins as natural scaffolds for protein engineering.

Vita

Roumestand

Toma

et al. 1995

Proc. Natl. Acad. Sci. U.S.A.

160

129

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A compact, well-organized, and natural motif, stabilized by three disulfide bonds, is proposed as a basic scaffold for protein engineering. This motif contains 37 amino acids only and is formed by a short helix on one face and an antiparallel triple-stranded 1l-sheet on the opposite face. It has been adopted by scorpions as a unique scaffold to express a wide variety of powerful toxic ligands with tuned specificity for different ion channels. We further tested the potential of this fold by engineering a metal binding site on it, taking the carbonic anhydrase site as a model. By chemical synthesis we introduced nine residues, including three histidines, as compared to the original amino acid sequence of the natural charybdotoxin and found that the new protein maintains the original fold, as revealed by CD and IH NMR analysis. Cu2+ ions are bound with Kd = 4.2 x 10-8 M and other metals are bound with affinities in an order mirroring that observed in carbonic anhydrase. The a/fl scorpion motif, small in size, easily amenable to chemical synthesis, highly stable, and tolerant for sequence mutations represents, therefore, an appropriate scaffold onto which polypeptide sequences may be introduced in a predetermined conformation, providing an additional 'means for design and engineering of small proteins.By engineering several disulfide bridges in short sequences (10-70 amino acids) nature has produced small proteins, toxins and protease inhibitors, that are able to adopt stable and biologically active structures. These small proteins bind to their biological targets with high affinity and specificity by virtue of their particular structure: an exposed loop fixed in a characteristic "canonical" conformation, which fits into a protease active site (e.g., in protease inhibitors; see refs. 1 and 2) or a more variegated molecular surface able to interact with a specific receptor or channel (e.g., in toxins; see refs. 3-5). In all these cases, the protein structure functions as a scaffold able to present a specific sequence and a determined fixed conformation to the biological target.These natural miniproteins constitute a priori interesting candidates as core structures for protein design, since the disulfide bonds provide most of their stabilization energy, leaving a large part of the protein structure available for mutations. In this respect, the natural motif of charybdotoxin (6-8) appears particularly well-suited; it is short (37 amino acids), it is composed of an antiparallel triple-stranded X3-sheet on one face and a short a-helix on the opposite face, and it is stabilized by three disulfide bonds in the interior core (8-10). The same fold is also adopted by (i) all known scorpion toxins (6-8), irrespective of their size, amino acid sequence, and function (blockage of K+ channels, Na+ channels, Cl-channels, etc.) (11-13); (ii) insect defensins (14); and (iii) plant y-thionins (15). The only residues commonly shared by all these proteins are the six cysteines involved in disulfide bonds (6, 7), indicating a hi...

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Section: Methodsmentioning

confidence: 99%

Scorpion toxins as natural scaffolds for protein engineering.

Vita

Roumestand

Toma

et al. 1995

Proc. Natl. Acad. Sci. U.S.A.

160

129

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“…Previously, we demonstrated flowcytometry-reactive antibodies to human CD4 in a high proportion (75 %) of cases [24]. We conclude that the physical characteristics of a given receptor peptide (e.g., length, hydrophilicity, etc.)…”

Section: Discussionmentioning

confidence: 51%

“…In previous work from this laboratory we demonstrated the induction of anti-receptor immunity using immunoglobulins (Ig) expressing discrete peptide portions of human CD4 [24]. We refer to such Ig as antigenized antibodies, i.e ., Ig molecules in which foreign peptide sequences are conformationally-constrained and expressed in the complementority-determining region (CDR) loops [25].…”

Section: Introductionmentioning

confidence: 99%

Antigenized antibodies expressing Vβ8.2 TCR peptides immunize against rat experimental allergic encephalomyelitis

Musselli

Daverio-Zanetti

Zanetti

2004

J Immune Based Ther Vaccines

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BackgroundImmunity against the T cell receptor (TCR) is considered to play a central role in the regulation of experimental allergic encephalomyelitis (EAE), a model system of autoimmune disease characterized by a restricted usage of TCR genes. Methods of specific vaccination against the TCR of pathogenetic T cells have included attenuated T cells and synthetic peptides from the sequence of the TCR. These approaches have led to the concept that anti-idiotypic immunity against antigenic sites of the TCR, which are a key regulatory element in this disease.MethodsThe present study in the Lewis rat used a conventional idiotypic immunization based on antigenized antibodies expressing selected peptide sequences of the Vβ8.2 TCR (93ASSDSSNTE101 and 39DMGHGLRLIHYSYDVNSTEKG59).ResultsThe study demonstrates that vaccination with antigenized antibodies markedly attenuates, and in some instances, prevents clinical EAE induced with the encephalitogenic peptide 68GSLPQKSQRSQDENPVVHF88 in complete Freunds' adjuvant (CFA). Antigenized antibodies induced an anti-idiotypic response against the Vβ8.2 TCR, which was detected by ELISA and flowcytometry. No evidence was obtained of a T cell response against the corresponding Vβ8.2 TCR peptides.ConclusionsThe results indicate that antigenized antibodies expressing conformationally-constrained TCR peptides are a simple means to induce humoral anti-idiotypic immunity against the TCR and to vaccinate against EAE. The study also suggests the possibility to target idiotypic determinants of TCR borne on pathogenetic T cells to vaccinate against disease.

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“…This provides the structural basis of IG variable domain antigenicity that could reside in the FR (public) or CDR (private), termed as idiotype. Indeed, viral T and B cell epitopes [37,38], parasitic B cell epitope [33] and CD4 molecule [39] have been used to antigenize IG that induced specific immune response subsequent to immunization. In this context, it is important to note that genetic imprints of all antigens present in the universe (10 8 −10 11 ) are enshrined within the framework of "internal mirror image" as idiotype in the antibody repertoire.…”

Section: Designing Antibody Fragments Of Desired Functionmentioning

confidence: 99%

Engineered Bovine Antibodies in the Development of Novel Therapeutics, Immunomodulators and Vaccines

et al. 2014

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Some bovine antibodies across all classes are unique, such as the CDR3 of the variable heavy-domain (VH CDR3), which is exceptionally long (up to 66 amino acids), unlike most conventional antibodies where the VH CDR3 loops range from 10 to 25 amino acids. The exceptionally long VH CDR3 is encoded by unusually long germline IGHD genes together with insertion of novel "a" nucleotide rich conserved short nucleotide sequence (CSNS) specifically at the IGH V-D junction. Such an exceptionally long VH CDR3 confers unique "knob and stalk" structural architecture where the knob, formed by intra-VH CDR3 disulfide bridges, is separated by 20 Å solvent exposed stalk composed of anti-parallel beta strands. The substitution of the knob with cytokines, such as, erythropoietin and granulocyte colony stimulating factor 3 (granulocyte colony stimulating factor), results in expression of functional fusion proteins with enhanced pharmacokinetics. The beta stranded stalk can be substituted with other rigid structures, for example, repeat alpha helices to form coiled-coil that mimics the beta-stranded stalk and, thus, opens opportunities for insertion of this structure in the CDRs of antibodies across species. Given the versatility of such a structural platform in bovine antibody VH CDR3, it provides the opportunity for the development of new generation of diagnostics, therapeutics, vaccines and immunomodulating drugs. OPEN ACCESS Antibodies 2014, 3 206

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Active immunity against the CD4 receptor by using an antibody antigenized with residues 41-55 of the first extracellular domain.

Cited by 17 publications

References 48 publications

Scorpion toxins as natural scaffolds for protein engineering.

Scorpion toxins as natural scaffolds for protein engineering.

Antigenized antibodies expressing Vβ8.2 TCR peptides immunize against rat experimental allergic encephalomyelitis

Engineered Bovine Antibodies in the Development of Novel Therapeutics, Immunomodulators and Vaccines

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