title: Drug specific antibodies: T-cell epitope-lipopeptide conjugates are potent adjuvants for small antigens in vivo and in vitro.
abstract: To generate conventional or monoclonal antibodies for the serological detection of drugs, antibiotics, toxins and other low molecular mass substances, a suitable and effective adjuvant is needed. Lipopeptides derived from a major component of the bacterial cell wall constitute potent nontoxic and nonpyrogenic immunoadjuvants when mixed with conventional antigens. Here we demonstrate that the synthetic lipopeptide N-palmitoyl-S-[2,3-bis(palmitoyloxy)-(2R,S)-propyl]-(R)-cysteinyl- serine (P3CS) coupled to a Th-cell epitope (P3CS-Th) can efficiently enhance the specific immune response against low molecular weight compounds in different species. In the presence of the synthetic lipopeptide P3CS-Th, the peptides which are per se non-immunogenic stimulated a specific humoral immune response in mice after intraperitoneal application. Mixtures containing adjuvants without the Th sequence showed no significant antibody induction. A marked enhancement of the humoral immune response was obtained with the low molecular mass antigens Iturin AL, Herbicolin A and Microcystin (MLR) coupled to poly-l-lysin (MLR-PLL), in rabbits and in chickens. Lipopeptide-Th cell epitope conjugates also constituted adjuvants for the in vitro immunization of either human mononuclear cells or mouse B-cells with MLR-PLL; after fusion of the immunized cultures with the heteromyeloma cell lines CB-F7 or the mouse myeloma cell line SP 2/0, respectively, we observed a significantly increased yield of antibody secreting hybridomas.
authors.affiliation: Institut für Immunbiologie der Universität, Freiburg, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Mittenbühler
authors.forename: M
authors.initials: M
authors.lastname: Loleit
authors.forename: W
authors.initials: W
authors.lastname: Baier
authors.forename: B
authors.initials: B
authors.lastname: Fischer
authors.forename: E
authors.initials: E
authors.lastname: Sedelmeier
authors.forename: G
authors.initials: G
authors.lastname: Jung
authors.forename: G
authors.initials: G
authors.lastname: Winkelmann
authors.forename: C
authors.initials: C
authors.lastname: Jacobi
authors.forename: J
authors.initials: J
authors.lastname: Weckesser
authors.forename: M H
authors.initials: MH
authors.lastname: Erhard
authors.forename: A
authors.initials: A
authors.lastname: Hofmann
authors.forename: W
authors.initials: W
authors.lastname: Bessler
authors.forename: P
authors.initials: P
authors.lastname: Hoffmann
mesh_terms: D000276:Adjuvants, Immunologic / Q000502:physiology*; D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D002645:Chickens; D004151:Dipeptides / Q000276:immunology; D004791:Enzyme Inhibitors / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007114:Immunization; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008074:Lipoproteins / Q000276:immunology / Q000378:metabolism*; D007959:Lymphocyte Culture Test, Mixed; D051379:Mice; D052998:Microcystins; D010455:Peptides / Q000276:immunology*; D010456:Peptides, Cyclic / Q000276:immunology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D011817:Rabbits
chemical_list: D000276:Adjuvants, Immunologic; D004151:Dipeptides; D004791:Enzyme Inhibitors; D018984:Epitopes, T-Lymphocyte; D008074:Lipoproteins; D052998:Microcystins; D010455:Peptides; D010456:Peptides, Cyclic; C078588:microcystin; C047210:2,3-bis-(palmitoyloxy)-2-propyl-N-palmitoyl-cysteinylserine; D010749:Phosphoprotein Phosphatases