title: Enhanced neutralization of HIV by antibodies displayed on the S-layer of Caulobacter crescentus.
abstract: Innovative methods of prevention are needed to stop the more than two million new HIV-1 infections annually, particularly in women. Local application of anti-HIV antibodies has been shown to be effective at preventing infection in nonhuman primates; however, the concentrations needed are cost prohibitive. Display of antibodies on a particulate platform will likely prolong effectiveness of these anti-HIV agents and lower the cost of goods. Here, we demonstrate that the bacterium Caulobacter crescentus and its highly expressed surface-layer (S-layer) protein can provide this antibody display platform. Caulobacters displaying protein G, alone or with CD4 codisplay, successfully captured HIV-1-specific antibodies and demonstrated functional neutralization. Compared to soluble antibodies, a neutralizing anti-HIV antibody displayed on Caulobacter was as effective or more effective at neutralizing diverse HIV-1 isolates. Moreover, when an antibody reactive with an epitope induced by CD4 binding (CD4i) was codisplayed with CD4, there was significant enhancement in HIV-1 neutralization. These results suggest that caulobacters displaying anti-HIV antibodies offer a distinct improvement in the use of antibodies as microbicides. Furthermore, these reagents can specifically evaluate anti-HIV antibodies in concert with other HIV-1 blocking agents to assess the most suitable tools for conversion to scFvs, allowing for direct display within the S-layer protein and further reducing cost of goods. In summary, C. crescentus, which can be easily produced and chemically stabilized at low cost, is well suited for engineering as an effective platform, offering an inexpensive way to produce and deliver HIV-1-specific microbicides.
authors: Department of Medicine, Beth Israel Deaconess Medical Center, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215, USA.
authors: Mark
authors: M
authors: Duval
authors: Christopher J
authors: CJ
authors: Lewis
authors: John F
authors: JF
authors: Nomellini
authors: Marc S
authors: MS
authors: Horwitz
authors: John
authors: J
authors: Smit
authors: Lisa A
authors: LA
authors: Cavacini
mesh_terms: D019380:Anti-HIV Agents / Q000276:immunology*; D015704:CD4 Antigens / Q000235:genetics / Q000378:metabolism; D016935:Caulobacter crescentus / Q000235:genetics / Q000378:metabolism*; D016503:Drug Delivery Systems; D005260:Female; D005818:Genetic Engineering / Q000379:methods; D015483:HIV Antibodies / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D015658:HIV Infections / Q000188:drug therapy / Q000276:immunology* / Q000821:virology; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D019380:Anti-HIV Agents; D015704:CD4 Antigens; D015483:HIV Antibodies; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; C055764:S-layer proteins