Potent neutralizing anti-CD1d antibody reduces lung cytokine release in primate asthma model

Abstract

(2015) Potent neutralizing anti-CD1d antibody reduces lung cytokine release in primate asthma model, mAbs, 7:3, 638-650, DOI: 10.1080/19420862.2015 To link to this article: https://doi.org/10. 1080/19420862.2015 Keywords: CD1d, NKT cell, antibody, asthma, cytokineAbbreviations: AHR, airway hyper-reactivity; APC, antigen-presenting cell; AUC, area under the curve; BAL, broncho-alveolar lavage; BSA, bovine serum albumin; CHO, Chinese hamster ovary; ELISA, enzyme-linked immunosorbent assay; G-CSF, granulocyte colony stimulating factor; GM-CSF, granulocyte-macrophage colony stimulating factor; HEK, human embryonic kidney; HPLC, high performance liquid chromatography; IFN, interferon; IL, interleukin; MCh, methacholine; MHC, major histocompatibility complex; MIP, macrophage inflammatory protein; NKT, natural killer T; OVA, ovalbumin; PBMC, peripheral blood mononuclear cell; PBS, phosphate-buffered saline; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SPR, surface plasmon resonance; TNF, tumor necrosis factor; VEGF, vascular endothelial growth factorCD1d is a receptor on antigen-presenting cells involved in triggering cell populations, particularly natural killer T (NKT) cells, to release high levels of cytokines. NKT cells are implicated in asthma pathology and blockade of the CD1d/ NKT cell pathway may have therapeutic potential. We developed a potent anti-human CD1d antibody (NIB.2) that possesses high affinity for human and cynomolgus macaque CD1d (K D »100 pM) and strong neutralizing activity in human primary cell-based assays (IC 50 typically <100 pM). By epitope mapping experiments, we showed that NIB.2 binds to CD1d in close proximity to the interface of CD1d and the Type 1 NKT cell receptor b-chain. Together with data showing that NIB.2 inhibited stimulation via CD1d loaded with different glycolipids, this supports a mechanism whereby NIB.2 inhibits NKT cell activation by inhibiting Type 1 NKT cell receptor b-chain interactions with CD1d, independent of the lipid antigen in the CD1d antigen-binding cleft. The strong in vitro potency of NIB.2 was reflected in vivo in an Ascaris suum cynomolgus macaque asthma model. Compared with vehicle control, NIB.2 treatment significantly reduced bronchoalveolar lavage (BAL) levels of Ascaris-induced cytokines IL-5, IL-8 and IL-1 receptor antagonist, and significantly reduced baseline levels of GM-CSF, IL-6, IL-15, IL-12/23p40, MIP-1a, MIP-1b, and VEGF. At a cellular population level NIB.2 also reduced numbers of BAL lymphocytes and macrophages, and blood eosinophils and basophils. We demonstrate that anti-CD1d antibody blockade of the CD1d/NKT pathway modulates inflammatory parameters in vivo in a primate inflammation model, with therapeutic potential for diseases where the local cytokine milieu is critical.

Generation and characterization of CD1d‐specific single‐domain antibodies with distinct functional features

“…Unique CD1d-specific VHH with the capacity to either stimulate moDC maturation and cytokine production, or to induce signs of early apoptosis in CD1d-expressing B lymphoblasts and MM cell lines, or to block the interaction between CD1d and iNKT cells were identified and characterized. Although mAbs with these features have been described, [21][22][23] this is the first report describing the generation of CD1d-specific VHH that are functional for one of the three mentioned processes.…”

“…In mouse, allergic asthma models showed decreased airway hyper‐reactivity and cytokine production in both CD1d‐deficient and iNKT‐deficient mice . Similarly, blockade of CD1d in a cynomolgus macaque airway hyper‐reactivity model reduced cytokine production and bronchial infiltration of lymphocytes and macrophages . As it was shown that airborne lipid antigens were able to induce profound inflammation via activation of iNKT cells in the lung, environmental lipids presented via CD1d have been implicated in the triggering and/or support of disease progression .…”

“…6 In line with these observations, both activation and prevention of iNKT activation have been reported to be able to positively affect disease outcome. Indeed, in a cynomolgus macaque asthma model, blocking of CD1d resulted in significantly reduced cytokine levels and lymphocyte infiltration, 23 indicating its therapeutic potential.…”

Cytotoxic lymphocytes and atherosclerosis: significance, mechanisms and therapeutic challenges

“…Recently, a CD1d lipid antagonist was shown to prevent iNKT cell activation in atherosclerotic mice and to reduce lesion inflammation and necrosis; the antagonist was also highly effective in preventing not only lesion development but also progression of established lesions (Li et al, ). Targeting antigen presentation with biologicals such as anti‐CD1d antibodies may also be an effective therapeutic strategy to prevent iNKT activation in atherosclerosis (Duthie et al, ); an anti‐human CD1d inhibitory antibody has recently been developed (Nambiar et al, ). Such approaches to limit activation of killer cells seem to impact on immune defence against infectious agents, but killer cells are able to respond against pathogens microbes via various innate receptors without utilizing TCR‐ or CD1d‐dependent activation.…”

The role of iNKT cells on the phenotypes of allergic airways in a mouse model

“…Studies in animals have suggested that iNKT cells might be important in allergic asthma, which has been supported by findings of their presence in the lung of patients with asthma [6, 21, 26–29], although some human studies do not support this conclusion [30–32]. In addition, crosstalk between iNKT cells and mast cells has been suggested to regulate inflammation [33].…”