Protection elicited by native outer membrane protein Oms66 (p66) against host-adapted Borrelia burgdorferi: conformational nature of bactericidal epitopes.
Oms66 is a Borrelia burgdorferi outer membrane porin protein whose role in Lyme disease pathogenesis and immunity has not been well established. Oms66 was solubilized from whole-cell lysates of strain B313 (which is derived from B31 but lacks OspA, -B, -C, and -D) and purified to homogeneity by fast-protein liquid chromatography. Purified native Oms66 (nOms66), which retained the ability to form large channels in a planar lipid bilayer model membrane system, and denatured Oms66 (hOms66) were used to immunize New Zealand White rabbits. The resulting Oms66 antisera were tested in a complement-dependent borreliacidal assay in parallel with basal serum and with serum from rabbits immune to reinfection with B. burgdorferi (IRS). IRS showed high-titer complement-dependent killing of both strains B31 and B313. Sera from animals immunized with nOms66 showed high-titer complement-dependent killing activity against strain B313 but exhibited no killing of B31. By comparison, serum generated from immunizations with hOms66 showed no killing activity against either strain. Following adsorption of antiserum to nOms66 with recombinant Oms66 (rOms66), the serum antibodies no longer bound to rOms66 or to nOms66 that had been denatured with 8 M urea. However, the antibodies still bound to nOms66 and killing activity against B313 was retained, thus suggesting that native, conformational epitopes are targets of this bactericidal activity. Six C3H HeJ mice were immunized with nOms66 and were challenged using "host-adapted" B. burgdorferi B31 by skin implantation of infected mouse ear tissue. Four of the six mice were protected against both localized and disseminated infection. These findings indicate that native Oms66 can elicit potent bactericidal activity and significant protective immunity against host-adapted organisms.
Exner
M M
MM
Division of Infectious Diseases, Department of Medicine, UCLA School of Medicine, Los Angeles, California 90095, USA. mexner@ucla.edu
Wu
X
X
Blanco
D R
DR
Miller
J N
JN
Lovett
M A
MA
D000222:Adaptation, Physiological; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D001426:Bacterial Proteins; D015748:Borrelia burgdorferi Group / Q000276:immunology*; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D015870:Gene Expression; D051379:Mice; D008809:Mice, Inbred C3H; D018272:Porins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011487:Protein Conformation; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D013987:Ticks / Q000382:microbiology; D014611:Vaccination
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D018985:Epitopes, B-Lymphocyte; C107893:Oms66 protein, Borrelia; D018272:Porins; D011993:Recombinant Fusion Proteins