Immunoreactivity of allergen (Bet v 1) conjugated to crystalline bacterial cell surface layers (S-layers).
BACKGROUND
Crystalline cell surface layers (S-layers) from Gram-positive eubacteria had been demonstrated as carrier/adjuvants for chemically synthesized tumor-associated oligosaccharide haptens and capsular polysaccharide antigens of Streptococcus pneumoniae strains.


OBJECTIVE
The applicability of S-layers as vaccine carrier for treatment of Type I allergy was investigated.


METHODS
Native or cross-linked S-layer self-assembly products and cell wall preparations from Bacillus sphaericus CCM 2177 and Thermoanaerobacter thermohydrosulfuricus L111-69 and L110-69 were used for immobilization of recombinant major birch pollen allergen Bet v 1.


CONCLUSIONS
Depending on the carrier used, amounts of approximately 20-40 micrograms allergen per mg conjugate could be immobilized. By application of L-glutamic acid dimethyl ester as a spacer, this value could be increased approximately 10-fold. The functionality of the rBet v 1-conjugates was assessed in immunological systems. (i) The presence of intact B-cell epitopes was demonstrated in inhibition experiments using human Bet v 1-specific IgE. (ii) The rBet v 1-S-layer conjugates were immunogenic in mice. (iii) The proliferation of rBet v 1-specific T-cell clones suggested that the peptides created by processing of immobilized Bet v 1 were similar to those derived from natural allergen. (iv) Stimulation of human allergen-specific TH2 lymphocytes with S-layer-conjugated Bet v 1 led to a modulation of the cytokine production pattern from TH2 to TH0/TH1. This study indicates that S-layers may be suitable carriers for few immunotherapeutical vaccines for Type 1 hypersensitivity.
Jahn-Schmid
B
B
Zentrum für Ultrastrukturforschung, Universität für Bodenkultur, Vienna, Austria.
Graninger
M
M
Glozik
M
M
Küpcü
S
S
Ebner
C
C
Unger
F M
FM
Sleytr
U B
UB
Messner
P
P
D000276:Adjuvants, Immunologic; D000485:Allergens / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D052179:Antigens, Plant; D001407:Bacillus / Q000737:chemistry / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology*; D002473:Cell Wall / Q000276:immunology; D003461:Crystallography; D016207:Cytokines / Q000096:biosynthesis; D000939:Epitopes; D005260:Female; D006094:Gram-Positive Bacteria / Q000737:chemistry / Q000276:immunology; D006801:Humans; D006967:Hypersensitivity / Q000628:therapy; D007073:Immunoglobulin E / Q000276:immunology; D007167:Immunotherapy; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D010940:Plant Proteins / Q000276:immunology*; D011058:Pollen / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology; D014197:Trees / Q000276:immunology; D014614:Vaccines, Synthetic / Q000737:chemistry
D000276:Adjuvants, Immunologic; D000485:Allergens; D052179:Antigens, Plant; D001426:Bacterial Proteins; D016207:Cytokines; D000939:Epitopes; D010940:Plant Proteins; D014614:Vaccines, Synthetic; C061312:Bet v 1 allergen, Betula; D007073:Immunoglobulin E