Inflammation and Immune Response
Inflammation is part of the lung’s natural immune response, however sometimes it is triggered when it shouldn’t be. If the lung tissue is inflamed, then this can lead to tissue scarring and breathing difficulties. The immune system in the lungs also plays an important role in the progression and pathology of a range of lung diseases, such as chronic obstructive pulmonary disorder, idiopathic pulmonary fibrosis (IPF) and asthma. 

The alveolar macrophage, a tissue-resident immune cell, plays a pivotal role in regulating normal alveolar function by serving as the first line of defence against inhaled particles and pathogens in the airways. Efficient functioning of these macrophages ensures regulated air exchange, essential for optimal respiratory health. On the other hand, a compromised alveolar macrophage response can disrupt normal alveolar function, obstructing air exchange and compromising lung health. Alveolar macrophages have the capability to produce signals that can either activate immune responses against threats or facilitate the healing process by resolving lung inflammation.

ImmuONE specialises in evaluating alveolar macrophage responses. We provide clients with a diverse array of in vitro alveolar macrophage assays to gauge cytokine activity, macrophage polarisation, phagocytic activity, and beyond. Our experts will support you in comprehending the variety of inflammatory and immunological responses that can manifest in the airways, utilising our in vitro models. Furthermore, we offer innovative in vitro experiments that give lung-relevant predictions of inflammatory responses. Some of the in vitro assessments we routinely conduct include:

Cytokine assessment

Cytokines are messenger molecules that initiate inflammatory immune responses. At ImmuONE, we offer services to measure the concentrations of cytokines and chemokines in macrophage cultures. This allows us to pinpoint and comprehend inflammatory reactions and associated signalling cascades. Utilising a suite of immuno-assay techniques – such as flow cytometry, membrane multiplexing and microarrays, and ELISA (Enzyme-Linked Immunosorbent Assay) – we design experiments tailored to your needs. Whether you’re aiming for a broad screen or a targeted assessment, we can evaluate single to multiple (from 1 to 50+ cytokines) concentrations seamlessly.

Macrophage Polarisation

Macrophages, depending on tissue-specific stimuli, can differentiate into two primary subtypes: M1 and M2. M1 macrophages are associated with pro-inflammatory activities, whereas M2 macrophages play roles in anti-inflammatory processes and tissue repair. At ImmuONE, we utilise our macrophage models to evaluate the distribution of these phenotypes within a given macrophage population. Furthermore, we have the capability to specifically induce M1 and M2 polarisation, enabling us to explore how various substances interact with activated or inflamed immune responses.

Nitric Oxide (NO) screening

Production of NO molecules is an important cytotoxic function that alveolar macrophages use to resolve infection. ImmuONE provides assays that determine the quantity of nitric oxide, which can serve as a general marker for the initiation of inflammation or an indicator of the level of M1 polarisation.

Irritation and Sensitisation

Irritation and sensitisation are adverse reactions to product exposure. Irritation is a short-term immediate response to product exposure whereas sensitisation is an inflammatory immune response that occurs after repeated exposure to a product. ImmuONE uses a combination approach of inflammatory, cytokine, biochemical and morphological endpoints to distinguish between respiratory irritation and sensitisation. We are developing in-house assays in this area. We also work with a range of other commercial partners with models and techniques to answer irritation and sensitisation needs. Whilst we specialise in lung irritation and sensitisation, we also work with customers addressing in vitro irritation and sensitisation questions in other tissues with tissue-resident macrophages present, such as the cornea.

Histological staining of in vitro lung tissues

We can perform histological staining of cells including cross-sectional preparation of tissues on cell culture inserts, which can be directly correlated to in vivo histology samples

Macrophage recovery

An important aspect of our research is understanding the phenomenon of ‘foamy macrophages’. These cells, evident by their vacuolated appearance, emerge in response to various stimuli. Such a phenotype can be induced by several factors including phospholipidosis, apoptosis, and other drug-related actions.

A significant concern in the inhalation medicine space is the ambiguity surrounding whether the vacuolated, or foamy, appearance indicates a temporary adaptive reaction or signifies a lasting detrimental change. We’ve harnessed advanced techniques like high content image analysis to achieve more detailed evaluations of these cells than what was possible with traditional light microscopy. Such advanced methods allow for intricate analysis of the cell’s vacuolation patterns, potentially shedding light on the mechanism of cellular response to different inhaled compounds.

Moreover, the persistence or reversibility of these changes is critical. We’ve observed varied responses in our studies. Some macrophages exhibit resilience post-exposure, suggesting adaptive changes, while others progress towards apoptosis, indicating adverse reactions.

At ImmuONE, we believe that a more detailed and nuanced understanding of these macrophage behaviours is pivotal in enhancing the development of inhaled medicines. By understanding whether a response is truly adverse or merely adaptive, better-informed decisions can be made during drug development, thereby ensuring the safety and efficacy of new therapies. Our expertise encompasses not only the observation and classification of these behaviours but extends to designing experiments to evaluate macrophage responses to single or multiple stimuli, enabling us to discern between adaptive and adverse responses.

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