title: Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus.
abstract: Eradication of human immunodeficiency virus type 1 (HIV-1) by vaccination with epitopes that produce broadly neutralizing antibodies is the ultimate goal for HIV prevention. However, generating appropriate immune responses has proven difficult. Expression of broadly neutralizing antibodies by vaginal colonizing lactobacilli provides an approach to passively target these antibodies to the mucosa. We tested the feasibility of expressing single-chain and single-domain antibodies (dAbs) in Lactobacillus to be used as a topical microbicide/live biotherapeutic. Lactobacilli provide an excellent platform to express anti-HIV proteins. Broadly neutralizing antibodies have been identified against epitopes on the HIV-1 envelope and have been made into active antibody fragments. We tested single-chain variable fragment m9 and dAb-m36 and its derivative m36.4 as prototype antibodies. We cloned and expressed the antibody fragments m9, m36, and m36.4 in Lactobacillus jensenii-1153 and tested the expression levels and functionality. We made a recombinant L. jensenii 1153-1128 that expresses dAb-m36.4. All antibody fragments m9, m36, and m36.4 were expressed by lactobacilli. However, we noted the smaller m36/m36.4 were expressed to higher levels, ≥3 μg/ml. All L. jensenii-expressed antibody fragments bound to gp120/CD4 complex; Lactobacillus-produced m36.4 inhibited HIV-1BaLin a neutralization assay. Using a TZM-bl assay, we characterized the breadth of neutralization of the m36.4. Delivery of dAbs by Lactobacillus could provide passive transfer of these antibodies to the mucosa and longevity at the site of HIV-1 transmission.
authors.affiliation: 1 Osel, Inc. , Mountain View, California.
authors.forename: Angela
authors.initials: A
authors.lastname: Marcobal
authors.affiliation: 1 Osel, Inc. , Mountain View, California.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Liu
authors.affiliation: 2 Profectus Biosciences, Inc. , Baltimore, Maryland.
authors.forename: Wenlei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: 2 Profectus Biosciences, Inc. , Baltimore, Maryland.
authors.forename: Antony S
authors.initials: AS
authors.lastname: Dimitrov
authors.affiliation: 1 Osel, Inc. , Mountain View, California.
authors.forename: Letong
authors.initials: L
authors.lastname: Jia
authors.affiliation: 1 Osel, Inc. , Mountain View, California.
authors.forename: Peter P
authors.initials: PP
authors.lastname: Lee
authors.affiliation: 2 Profectus Biosciences, Inc. , Baltimore, Maryland.
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Fouts
authors.affiliation: 1 Osel, Inc. , Mountain View, California.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Parks
authors.affiliation: 1 Osel, Inc. , Mountain View, California.
authors.forename: Laurel A
authors.initials: LA
authors.lastname: Lagenaur
mesh_terms: D057134:Antibodies, Neutralizing / Q000235:genetics / Q000276:immunology*; D018562:Disease Transmission, Infectious / Q000517:prevention & control; D005260:Female; D015870:Gene Expression; D015483:HIV Antibodies / Q000235:genetics / Q000276:immunology*; D015658:HIV Infections / Q000517:prevention & control; D015497:HIV-1; D006801:Humans; D007778:Lactobacillus / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D014621:Vagina / Q000276:immunology / Q000382:microbiology
chemical_list: D057134:Antibodies, Neutralizing; D015483:HIV Antibodies; D011994:Recombinant Proteins
keywords: HIV neutralization; Lactobacillus; passive delivery; single-domain antibody