Vaccination with ADCC activating HA peptide epitopes provides partial protection from influenza infection

Abstract

Influenza-specific antibody dependent cellular cytotoxicity (ADCC) antibodies have a broad cross reactivity and potential as an immune correlate for universal vaccines. Peptide-mapping for ADCC reactivity of H1-HA and H7-HA proteins from human serum samples identified high ADCC-inducing peptides in both the HA1 and HA2 regions. Vaccination of mice with single ADCC-peptides induced ADCC activity leading to partial protection from lethal influenza challenge, with increased survival, reduced viral loads, and reduced activation of NK cells in the lungs. Targeted vaccination strategies to elicit ADCC responses may provide an approach for universal vaccines.

The Role of Fc Receptors on the Effectiveness of Therapeutic Monoclonal Antibodies

“…Effector cell cultures for such assays can comprise crude PBMC cell cultures, primary NK cells, or NK cell lines. NK cell-mediated killing upon α-CD20:FcγRIIIa crosslinking can be detected via released labeling components in the cell culture supernatant (pre-labeled 51 Cr, fluorescent dyes, or release of internal enzymes such as lactate dehydrogenase (LDH)). Further ADCC read outs are the detection of viability via FACS-based analyses or determining degranulation (CD107 by FACS analyses or ELISA).…”

“…For example, WIL2-S cells, a human B-lymphoblastic cell line (American Type Culture Collection (ATCC), CRL-8885), and Raji cells or Daudi cells both derived from a Burkitt's lymphoma (Raji: ATCC, CCL-86; Daudi: ATCC, CCL-213) are applied in the assessment of Fc-mediated effector function. In addition, primary cells obtained from CLL patients can be used as target cells for CD20-specific mAbs [43,[46][47][48][49][50][51].…”

“…The usage of cell-aliquots from one blood donor could facilitate minimization of variations. Another approach to circumvent donor-dependent variations, is the establishment of NK cell lines as effector cells such as NK-92 variants and YT-CD16 cells [30,46,48,51].…”

“…The readout of the ADCC assays is mostly defined by the specific lysis of the target cells, using respective controls of full lysis (e.g., Triton-X 100 treatment), unspecific lysis of co-cultures (without addition of the therapeutic mAb), and taking spontaneous lyses of untreated cells into account [30,46,48]. Determination of lysis was measured via 51 Crrelease by target cells for many years [53,56]. Nowadays, such analyses can be based on the decrease of fluorescently labeled target cells (e.g., Cell-Trace Violet), increase of relative fluorescence units (RFU) in cell culture supernatants (e.g., Calcein-AM [46,52]), by measuring lactate dehydrogenase (LDH)-release [47,48,54], or using specific labeling systems [30,48].…”

“…VanDerMeid et al defined CLL target-cells as viable, if they were negative for both Annexin V and a dye that only penetrates dying cells [50]. Mata et al also replaced the 51 Cr release assay by a FACS-based assay to assess ADCC. The detection of CD107 (lysosomal-associated membrane glycoprotein-1, LAMP-1) as a marker for degranulation, perforin tracking, and granule movement [57] was used/established as surrogate marker for activation of NK effector cells [53].…”