Biology subjects

Madhireddy, D.

Publications and source records attributed to Madhireddy, D..

3 recordsLinked to original sources

bioRxiv2019

Cell type-specific monoclonal antibody cross-reactivity screening in non-human primates and development of comparative immunophenotyping panels for CyTOF

Monoclonal antibodies are a critical tool for immunologists, with applications including immunomicroscopy and cytometry, and non-human primates are essential models for drug development. Very few antibodies are raised against non-human primate antigens; instead, researchers typically use anti-human antibodies that are found to be cross-reactive with non-human primates. The NIH maintains a valuable database of cross-reactivity, but its coverage is not complete, especially for less common species. Furthermore, the database only indicates the presence or absence of staining and does not indicate if a different cell population is stained in the NHP species than in humans. We screened 332 antibodies in five immune cell populations in blood from cynomologus macaques (Macaca fascicularis), rhesus macaques (Macaca mulatta), African green monkeys (Chlorocebus aethiops) and olive/yellow baboons (Papio hamadryas anubis x Papio hamadryas cynocephalus), thereby generating a comprehensive cross-reactivity catalog that includes cell type-specificity. We subsequently used this catalog to help create large CyTOF mass cytometry panels for three of those species and humans. The curated dataset containing the primary data for each antibody has been deposited as a browsable resource at https://immuneatlas.org and https://flowrepository.org/id/FR-FCM-Z2Z7.

immunology↗

bioRxiv2019

A comprehensive atlas of immunological differences between humans, mice and non-human primates

Animal models are an integral part of the drug development and evaluation process. However, they are unsurprisingly imperfect reflections of humans, and the extent and nature of many immunological differences are unknown. With the rise of targeted and biological therapeutics, it is increasingly important that we understand the molecular differences in immunological behavior of humans and model organisms. Thus, we profiled a large number of healthy humans, along with three of the model organisms most similar to humans: rhesus and cynomolgus macaques and African green monkeys; and the most widely used mammalian model: mice. Using cross-species, universal phenotyping and signaling panels, we measured immune cell signaling responses to an array of 15 stimuli using CyTOF mass cytometry. We found numerous instances of different cellular phenotypes and immune signaling events occurring within and between species with likely effects on evaluation of therapeutics, and detail three examples (double-positive T cell frequency and signaling; granulocyte response to Bacillus anthracis antigen; and B cell subsets). We also explore the correlation of herpes simian B virus serostatus on the immune profile. The full dataset is available online at https://flowrepository.org (accession FR-FCM-Z2ZY) and https://immuneatlas.org.

immunology↗

bioRxiv2019

Variation of immune cell responses in humans reveals sex-specific coordinated signaling across cell types

Assessing the health and competence of the immune system is central to evaluating vaccination responses, autoimmune conditions, cancer prognosis and treatment. With an increasing number of studies examining immune dysregulation, there is a growing need for a curated reference of variation in immune parameters in healthy individuals. We used mass cytometry (CyTOF) to profile blood from 86 humans in response to 15 ex vivo immune stimuli. We present reference ranges for cell-specific immune markers and highlight differences that appear across sex and age. We identified modules of immune features that suggests there exists and underlying structure to the immune system based on signaling pathway responses across cell types. We observed increased MAPK signaling in inflammatory pathways in innate immune cells and greater overall coordination of immune cell responses in women. In contrast, men exhibited stronger STAT1 and TBK1 responses. These reference data are publicly available as a resource for immune profiling studies.

immunology↗