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Event: 151

Key Event Title

The KE title should describe a discrete biological change that can be measured. It should generally define the biological object or process being measured and whether it is increased, decreased, or otherwise definably altered relative to a control state. For example “enzyme activity, decreased”, “hormone concentration, increased”, or “growth rate, decreased”, where the specific enzyme or hormone being measured is defined. More help

Activation, Inflammatory cytokines, chemokines, cytoprotective gene pathways

Short name
The KE short name should be a reasonable abbreviation of the KE title and is used in labelling this object throughout the AOP-Wiki. The short name should be less than 80 characters in length. More help
Activation, Inflammatory cytokines, chemokines, cytoprotective gene pathways

Biological Context

Structured terms, selected from a drop-down menu, are used to identify the level of biological organization for each KE. Note, KEs should be defined within a particular level of biological organization. Only KERs should be used to transition from one level of organization to another. Selection of the level of biological organization defines which structured terms will be available to select when defining the Event Components (below). More help
Level of Biological Organization

Cell term

Further information on Event Components and Biological Context may be viewed on the attached pdf.The biological context describes the location/biological environment in which the event takes place.  For molecular/cellular events this would include the cellular context (if known), organ context, and species/life stage/sex for which the event is relevant. For tissue/organ events cellular context is not applicable.  For individual/population events, the organ context is not applicable. More help
Cell term
eukaryotic cell

Organ term

Further information on Event Components and Biological Context may be viewed on the attached pdf.The biological context describes the location/biological environment in which the event takes place.  For molecular/cellular events this would include the cellular context (if known), organ context, and species/life stage/sex for which the event is relevant. For tissue/organ events cellular context is not applicable.  For individual/population events, the organ context is not applicable. More help

Key Event Components

Further information on Event Components and Biological Context may be viewed on the attached pdf.Because one of the aims of the AOP-KB is to facilitate de facto construction of AOP networks through the use of shared KE and KER elements, authors are also asked to define their KEs using a set of structured ontology terms (Event Components). In the absence of structured terms, the same KE can readily be defined using a number of synonymous titles (read by a computer as character strings). In order to make these synonymous KEs more machine-readable, KEs should also be defined by one or more “event components” consisting of a biological process, object, and action with each term originating from one of 22 biological ontologies (Ives, et al., 2017; See List). Biological process describes dynamics of the underlying biological system (e.g., receptor signalling). The biological object is the subject of the perturbation (e.g., a specific biological receptor that is activated or inhibited). Action represents the direction of perturbation of this system (generally increased or decreased; e.g., ‘decreased’ in the case of a receptor that is inhibited to indicate a decrease in the signalling by that receptor).Note that when editing Event Components, clicking an existing Event Component from the Suggestions menu will autopopulate these fields, along with their source ID and description. To clear any fields before submitting the event component, use the 'Clear process,' 'Clear object,' or 'Clear action' buttons. If a desired term does not exist, a new term request may be made via Term Requests. Event components may not be edited; to edit an event component, remove the existing event component and create a new one using the terms that you wish to add. More help
Process Object Action
chemokine activity Chemokine increased
cytokine activity Cytokine increased

Key Event Overview

AOPs Including This Key Event

All of the AOPs that are linked to this KE will automatically be listed in this subsection. This table can be particularly useful for derivation of AOP networks including the KE. Clicking on the name of the AOP will bring you to the individual page for that AOP. More help
AOP Name Role of event in AOP Point of Contact Author Status OECD Status
Covalent binding to proteins leads to Respiratory Sensitisation/Sensitization/Allergy KeyEvent Kristie Sullivan (send email) Under Development: Contributions and Comments Welcome Under Development


This is a structured field used to identify specific agents (generally chemicals) that can trigger the KE. Stressors identified in this field will be linked to the KE in a machine-readable manner, such that, for example, a stressor search would identify this as an event the stressor can trigger. NOTE: intermediate or downstream KEs in one AOP may function as MIEs in other AOPs, meaning that stressor information may be added to the KE description, even if it is a downstream KE in the pathway currently under development.Information concerning the stressors that may trigger an MIE can be defined using a combination of structured and unstructured (free-text) fields. For example, structured fields may be used to indicate specific chemicals for which there is evidence of an interaction relevant to this MIE. By linking the KE description to a structured chemical name, it will be increasingly possible to link the MIE to other sources of chemical data and information, enhancing searchability and inter-operability among different data-sources and knowledgebases. The free-text section “Evidence for perturbation of this MIE by stressor” can be used both to identify the supporting evidence for specific stressors triggering the MIE as well as to define broad chemical categories or other properties that classify the stressors able to trigger the MIE for which specific structured terms may not exist. More help

Taxonomic Applicability

Latin or common names of a species or broader taxonomic grouping (e.g., class, order, family) can be selected from an ontology. In many cases, individual species identified in these structured fields will be those for which the strongest evidence used in constructing the AOP was available in relation to this KE. More help
Term Scientific Term Evidence Link
human Homo sapiens Moderate NCBI

Life Stages

The structured ontology terms for life-stage are more comprehensive than those for taxa, but may still require further description/development and explanation in the free text section. More help
Life stage Evidence
All life stages

Sex Applicability

No help message More help
Term Evidence

Key Event Description

A description of the biological state being observed or measured, the biological compartment in which it is measured, and its general role in the biology should be provided. For example, the biological state being measured could be the activity of an enzyme, the expression of a gene or abundance of an mRNA transcript, the concentration of a hormone or protein, neuronal activity, heart rate, etc. The biological compartment may be a particular cell type, tissue, organ, fluid (e.g., plasma, cerebrospinal fluid), etc. The role in the biology could describe the reaction that an enzyme catalyses and the role of that reaction within a given metabolic pathway; the protein that a gene or mRNA transcript codes for and the function of that protein; the function of a hormone in a given target tissue, physiological function of an organ, etc. Careful attention should be taken to avoid reference to other KEs, KERs or AOPs. Only describe this KE as a single isolated measurable event/state. This will ensure that the KE is modular and can be used by other AOPs, thereby facilitating construction of AOP networks. More help

The innate immune system plays a crucial role in the initiation of adaptive immune responses. (Poynter, 2012, Salazar and Ghaemmaghami, 2013) It is a first-line of defense against invading microbial pathogens and is activated via a range of pattern recognition receptors (PRRs) that recognize conserved patterns present on pathogens, that is, the toll-like receptors (TLRs) and the nucleotide binding domain leucine-rich repeat containing receptor (NLR) family. These PRRs can be activated by endogenous danger-associated molecular patterns (DAMPs), released under oxidative stress and cell damage and include components of the extracellular matrix generated after tissue injury, for example, hyaluronic acid fragments, intracellular proteins such as heat shock proteins and nonprotein DAMPs such as uric acid crystals. (Kawai and Akira, 2010, Seong and Matzinger, 2004, Wheeler et al., 2009)

NLR protein-3 (NLRP3) is a PRR that belongs to the NLR family, a group of intracellular receptors activated by mitochondrial oxidative stress, for example, by adenosine triphosphate  and uric acid. (Kawai and Akira, 2009) On activation, TLR and NLRP3 activate innate immunity signaling pathways leading to the release of proinflammatory cytokines and chemokines. In recent years, increasing attention has been paid to the role of the innate immune system in asthma. The sentinel role of the innate immune systems includes the activation of pathways by pathogen-associated molecular patterns and DAMPs. By this, KEs during sensitization such as activation and migration of DCs are set into motion. (Holgate, 2012) Proinflammatory molecules are also known to induce the expression of surface molecules on immune cells such as antigen-presenting cells (APCs), which are greatly involved in the induction of adaptive immune responses. Thus, whether an immune response or tolerance response is induced in APCs depends not only on the presence of antigenic properties of a substance but also on danger signals.

How It Is Measured or Detected

One of the primary considerations in evaluating AOPs is the relevance and reliability of the methods with which the KEs can be measured. The aim of this section of the KE description is not to provide detailed protocols, but rather to capture, in a sentence or two, per method, the type(s) of measurements that can be employed to evaluate the KE and the relative level of scientific confidence in those measurements. Methods that can be used to detect or measure the biological state represented in the KE should be briefly described and/or cited. These can range from citation of specific validated test guidelines, citation of specific methods published in the peer reviewed literature, or outlines of a general protocol or approach (e.g., a protein may be measured by ELISA).Key considerations regarding scientific confidence in the measurement approach include whether the assay is fit for purpose, whether it provides a direct or indirect measure of the biological state in question, whether it is repeatable and reproducible, and the extent to which it is accepted in the scientific and/or regulatory community. Information can be obtained from the OECD Test Guidelines website and the EURL ECVAM Database Service on Alternative Methods to Animal Experimentation (DB-ALM). ?

There are no predictive markers for cellular danger or proinflammatory responses described for respiratory sensitizers yet. The studies performed up until now did not result in any proteins, genes, or molecular pathways that are consistently regulated by a broad range of respiratory sensitizers or genes; (Remy et al., 2014) however, only a few chemicals have been tested. Cytokine production can be measured by ELISA or Bio-Plex systems either in the supernatants or intracellular matrix. Cell systems that can be used include also complex models such as the 3D epithelial cell models, that is, MucilAir™ and PCLS. (Huang et al., 2013, Lauenstein et al., 2014)

Activation of innate immune response can also be assessed using commercial immunoassays for signal transduction pathways, that is, p38 MAPK, JNK 1/2, and ERK 1/2. Other possible detection methods, focusing on ROS production or the induction of cytoprotective pathways, might be used as well to assess the ability of chemicals to generate endogenous danger signals (DAMPs). For ROS production, commercial assays are available that can be applied. The induction of Nrf2-KEAP1 can be assessed using the Keratinosens® (Natsch et al., 2013, Emter et al., 2010) or LuSens (Ramirez et al., 2014) assays (OECD TG 442D) and by measuring gene expression of Nrf2-dependent genes by quantitative polymerase chain reaction (qPCR), that is, HMOX, (Migdal et al., 2013) although the utility of this pathway for respiratory sensitizers is unclear. The BEAS-2B cell line, coupled with microarray analysis, reveals the PTEN pathway as potentially useful. (Verstraelen et al., 2009) The predictivity of these assays has not been studied with a large number of respiratory sensitizers.

Domain of Applicability

This free text section should be used to elaborate on the scientific basis for the indicated domains of applicability and the WoE calls (if provided). While structured terms may be selected to define the taxonomic, life stage and sex applicability (see structured applicability terms, above) of the KE, the structured terms may not adequately reflect or capture the overall biological applicability domain (particularly with regard to taxa). Likewise, the structured terms do not provide an explanation or rationale for the selection. The free-text section on evidence for taxonomic, life stage, and sex applicability can be used to elaborate on why the specific structured terms were selected, and provide supporting references and background information.  More help

It is not fully understood which cell types are the most important sources for the endogenous danger signals involved in sensitization of the respiratory tract. Relevant cell types representing cellular sources for danger signals are probably alveolar and bronchial epithelial cells, keratinocytes, macrophages, DCs, natural killer cells, endothelial cells, and nerve fiber endings. (Verstraelen et al., 2008) In particular, macrophages are able to respond with high levels of, for example, cytokines and ROS after stimulation of PRRs. Human cell lines representative of the cells mentioned above might be used for the measurements of danger signal induction. A limitation of the use of submerged cell lines is that certain respiratory sensitizers hydrolyze in an aqueous environment, which may lead to negative results. (Wanner et al., 2010) Air/liquid exposure in 3D skin or airway models might provide a more robust model although this has not been explored in great detail.


List of the literature that was cited for this KE description. Ideally, the list of references, should conform, to the extent possible, with the OECD Style Guide ( (OECD, 2015). More help

EMTER, R., ELLIS, G. & NATSCH, A. 2010. Performance of a novel keratinocyte-based reporter cell line to screen skin sensitizers in vitro. Toxicol Appl Pharmacol, 245, 281-90.

HOLGATE, S. T. 2012. Innate and adaptive immune responses in asthma. Nat Med, 18, 673-83.

HUANG, S., WISZNIEWSKI, L., CONSTANT, S. & ROGGEN, E. 2013. Potential of in vitro reconstituted 3D human airway epithelia (MucilAir™) to assess respiratory sensitizers. Toxicol In Vitro, 27, 1151-6.

KAWAI, T. & AKIRA, S. 2009. The roles of TLRs, RLRs and NLRs in pathogen recognition. Int Immunol, 21, 317-37.

KAWAI, T. & AKIRA, S. 2010. The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors. Nat Immunol, 11, 373-84.

LAUENSTEIN, L., SWITALLA, S., PRENZLER, F., SEEHASE, S., PFENNIG, O., FÖRSTER, C., FIEGUTH, H., BRAUN, A. & SEWALD, K. 2014. Assessment of immunotoxicity induced by chemicals in human precision-cut lung slices (PCLS). Toxicol In Vitro, 28, 588-99.

MIGDAL, C., BOTTON, J., EL ALI, Z., AZOURY, M. E., GULDEMANN, J., GIMÉNEZ-ARNAU, E., LEPOITTEVIN, J. P., KERDINE-RÖMER, S. & PALLARDY, M. 2013. Reactivity of chemical sensitizers toward amino acids in cellulo plays a role in the activation of the Nrf2-ARE pathway in human monocyte dendritic cells and the THP-1 cell line. Toxicol Sci, 133, 259-74.

NATSCH, A., RYAN, C. A., FOERTSCH, L., EMTER, R., JAWORSKA, J., GERBERICK, F. & KERN, P. 2013. A dataset on 145 chemicals tested in alternative assays for skin sensitization undergoing prevalidation. J Appl Toxicol, 33, 1337-52.

POYNTER, M. E. 2012. Airway epithelial regulation of allergic sensitization in asthma. Pulm Pharmacol Ther, 25, 438-46.

RAMIREZ, T., MEHLING, A., KOLLE, S. N., WRUCK, C. J., TEUBNER, W., ELTZE, T., AUMANN, A., URBISCH, D., VAN RAVENZWAAY, B. & LANDSIEDEL, R. 2014. LuSens: a keratinocyte based ARE reporter gene assay for use in integrated testing strategies for skin sensitization hazard identification. Toxicol In Vitro, 28, 1482-97.

REMY, S., VERSTRAELEN, S., VAN DEN HEUVEL, R., NELISSEN, I., LAMBRECHTS, N., HOOYBERGHS, J. & SCHOETERS, G. 2014. Gene expressions changes in bronchial epithelial cells: markers for respiratory sensitizers and exploration of the NRF2 pathway. Toxicol In Vitro, 28, 209-17.

SALAZAR, F. & GHAEMMAGHAMI, A. M. 2013. Allergen recognition by innate immune cells: critical role of dendritic and epithelial cells. Front Immunol, 4, 356.

SEONG, S. Y. & MATZINGER, P. 2004. Hydrophobicity: an ancient damage-associated molecular pattern that initiates innate immune responses. Nat Rev Immunol, 4, 469-78.

VERSTRAELEN, S., BLOEMEN, K., NELISSEN, I., WITTERS, H., SCHOETERS, G. & VAN DEN HEUVEL, R. 2008. Cell types involved in allergic asthma and their use in in vitro models to assess respiratory sensitization. Toxicol In Vitro, 22, 1419-31.

VERSTRAELEN, S., NELISSEN, I., HOOYBERGHS, J., WITTERS, H., SCHOETERS, G., VAN CAUWENBERGE, P. & VAN DEN HEUVEL, R. 2009. Gene profiles of a human alveolar epithelial cell line after in vitro exposure to respiratory (non-)sensitizing chemicals: identification of discriminating genetic markers and pathway analysis. Toxicol Lett, 185, 16-22.

WANNER, R., SONNENBURG, A., QUATCHADZE, M., SCHREINER, M., PEISER, M., ZUBERBIER, T. & STAHLMANN, R. 2010. Classification of sensitizing and irritative potential in a combined in-vitro assay. Toxicol Appl Pharmacol, 245, 211-8.

WHEELER, D. S., CHASE, M. A., SENFT, A. P., POYNTER, S. E., WONG, H. R. & PAGE, K. 2009. Extracellular Hsp72, an endogenous DAMP, is released by virally infected airway epithelial cells and activates neutrophils via Toll-like receptor (TLR)-4. Respir Res, 10, 31.