title: Adjuvant-induced Human Monocyte Secretome Profiles Reveal Adjuvant- and Age-specific Protein Signatures.
abstract: Adjuvants boost vaccine responses, enhancing protective immunity against infections that are most common among the very young. Many adjuvants activate innate immunity, some via Toll-Like Receptors (TLRs), whose activities varies with age. Accordingly, characterization of age-specific adjuvant-induced immune responses may inform rational adjuvant design targeting vulnerable populations. In this study, we employed proteomics to characterize the adjuvant-induced changes of secretomes from human newborn and adult monocytes in response to Alum, the most commonly used adjuvant in licensed vaccines; Monophosphoryl Lipid A (MPLA), a TLR4-activating adjuvant component of a licensed Human Papilloma Virus vaccine; and R848 an imidazoquinoline TLR7/8 agonist that is a candidate adjuvant for early life vaccines. Monocytes were incubated in vitro for 24 h with vehicle, Alum, MPLA, or R848 and supernatants collected for proteomic analysis employing liquid chromatography-mass spectrometry (LC-MS) (data available via ProteomeXchange, ID PXD003534). 1894 non-redundant proteins were identified, of which ∼30 - 40% were common to all treatment conditions and ∼5% were treatment-specific. Adjuvant-stimulated secretome profiles, as identified by cluster analyses of over-represented proteins, varied with age and adjuvant type. Adjuvants, especially Alum, activated multiple innate immune pathways as assessed by functional enrichment analyses. Release of lactoferrin, pentraxin 3, and matrix metalloproteinase-9 was confirmed in newborn and adult whole blood and blood monocytes stimulated with adjuvants alone or adjuvanted licensed vaccines with distinct clinical reactogenicity profiles. MPLA-induced adult monocyte secretome profiles correlated in silico with transcriptome profiles induced in adults immunized with the MPLA-adjuvanted RTS,S malaria vaccine (Mosquirix™). Overall, adjuvants such as Alum, MPLA and R848 give rise to distinct and age-specific monocyte secretome profiles, paralleling responses to adjuvant-containing vaccines in vivo Age-specific in vitro modeling coupled with proteomics may provide fresh insight into the ontogeny of adjuvant action thereby informing targeted adjuvanted vaccine development for distinct age groups.
authors.affiliation: From the ‡Department of Medicine, Division of Infectious Diseases, Boston Children's Hospital and §Harvard Medical School, Boston, Massachusetts; ¶Division of Pediatric Infectious Diseases, New York University Medical School, New York; Precision Vaccines Program, Division of Infectious Diseases, Boston Children's Hospital, Boston, Massachusetts.
authors.forename: Djin-Ye
authors.initials: DY
authors.lastname: Oh
authors.affiliation: From the ‡Department of Medicine, Division of Infectious Diseases, Boston Children's Hospital and §Harvard Medical School, Boston, Massachusetts;
authors.forename: David J
authors.initials: DJ
authors.lastname: Dowling
authors.affiliation: ‖Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts;
authors.forename: Saima
authors.initials: S
authors.lastname: Ahmed
authors.affiliation: **Saw Swee Hock School of Public Health, National University of Singapore, Singapore;
authors.forename: Hyungwon
authors.initials: H
authors.lastname: Choi
authors.affiliation: From the ‡Department of Medicine, Division of Infectious Diseases, Boston Children's Hospital and.
authors.forename: Spencer
authors.initials: S
authors.lastname: Brightman
authors.affiliation: From the ‡Department of Medicine, Division of Infectious Diseases, Boston Children's Hospital and.
authors.forename: Ilana
authors.initials: I
authors.lastname: Bergelson
authors.affiliation: ‖Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts;
authors.forename: Sebastian T
authors.initials: ST
authors.lastname: Berger
authors.affiliation: ‖Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts;
authors.forename: John F
authors.initials: JF
authors.lastname: Sauld
authors.affiliation: From the ‡Department of Medicine, Division of Infectious Diseases, Boston Children's Hospital and §Harvard Medical School, Boston, Massachusetts; Precision Vaccines Program, Division of Infectious Diseases, Boston Children's Hospital, Boston, Massachusetts.
authors.forename: Matthew
authors.initials: M
authors.lastname: Pettengill
authors.affiliation: ‡‡Children's Hospital Informatics Program, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts;
authors.forename: Alvin T
authors.initials: AT
authors.lastname: Kho
authors.affiliation: ¶Division of Pediatric Infectious Diseases, New York University Medical School, New York;
authors.forename: Henry J
authors.initials: HJ
authors.lastname: Pollack
authors.affiliation: ‖Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts; Precision Vaccines Program, Division of Infectious Diseases, Boston Children's Hospital, Boston, Massachusetts.
authors.forename: Hanno
authors.initials: H
authors.lastname: Steen
authors.affiliation: From the ‡Department of Medicine, Division of Infectious Diseases, Boston Children's Hospital and §Harvard Medical School, Boston, Massachusetts; Precision Vaccines Program, Division of Infectious Diseases, Boston Children's Hospital, Boston, Massachusetts ofer.levy@childrens.harvard.edu.
authors.forename: Ofer
authors.initials: O
authors.lastname: Levy
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000328:Adult; D000367:Age Factors; D000534:Alum Compounds / Q000494:pharmacology; D002853:Chromatography, Liquid; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007113:Immunity, Innate / Q000187:drug effects; D007231:Infant, Newborn; D008050:Lipid A / Q000031:analogs & derivatives / Q000494:pharmacology; D013058:Mass Spectrometry; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism; D020543:Proteome / Q000187:drug effects / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*
chemical_list: D000276:Adjuvants, Immunologic; D000534:Alum Compounds; D007093:Imidazoles; D008050:Lipid A; D020543:Proteome; C041524:aluminum sulfate; C048436:monophosphoryl lipid A; C402365:resiquimod