Rational design of synthetic peptides to generate antibodies that recognize in situ CD11c(+) putative dendritic cells in horse lymph nodes.
A three-dimensional model of the alphaX I-domain of the horse integrin CD11c from dendritic cells provided information for selecting two segments of the primary structure for peptide synthesis. Peptide 1 contains 20 amino acids and peptide 2 has 17 amino acid residues. The first spans from position Thr229 to Arg248 of an alpha-helix segment of the structure, whereas peptide 2 goes from Asp158 to Phe174 and corresponds to an exposed segment of the loop considered to be the metal ion-dependent adhesion site. Murine polyclonal antisera against both peptides were generated and assayed in peripheral blood cell suspensions and in cryosections of horse lymph nodes. Only the serum against peptide 2 was capable of identifying cells in suspension and in situ by immunohistochemistry, some with evident dendritic morphology. Using this approach, an immunogenic epitope exposed in CD11c was identified in cells from horse lymph node in situ.
Espino-Solis
Gerardo P
GP
Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, 62210 Morelos, Mexico.
Calderon-Amador
J
J
Calderon-Aranda
E S
ES
Licea
A F
AF
Donis-Maturano
L
L
Flores-Romo
L
L
Possani
L D
LD
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D039521:CD11c Antigen / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D003429:Cross Reactions; D003713:Dendritic Cells / Q000276:immunology*; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D005260:Female; D006736:Horses / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007150:Immunohistochemistry; D008198:Lymph Nodes / Q000166:cytology / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D015202:Protein Engineering; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid
D039521:CD11c Antigen; D000939:Epitopes; D010446:Peptide Fragments