Expression of the Memory Marker CD45RO on Helper T Cells in Macaques

Valentine, Michael; Song, Kejing; Maresh, Grace A.; Mack, Heather; Huaman, Maria Cecilia; Polacino, Patricia; Ho, On; Cristillo, Anthony D.; Chung, Hye Kyung; Hu, Shiu Lok; Pincus, Seth H.

doi:10.1371/journal.pone.0073969

Cited by 11 publications

(5 citation statements)

References 37 publications

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“…Therefore, we primarily use CD16 in lieu of CD11c during gating analysis for consistency. In the case of naïve and memory T cells, we observed a very broad distribution of CD45RA staining, as previously reported ( 34 , 35 ), with two clones (5H9 and HI100), which was difficult to gate (see online dataset). Staining of this marker was superior with the VersaLyse-based method than with the Triton-X100-based method (see Materials and Methods ) (data not shown).…”

Section: Resultssupporting

confidence: 84%

A Comprehensive Atlas of Immunological Differences Between Humans, Mice, and Non-Human Primates

et al. 2022

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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 the immunological behavior of humans and model organisms. However, very few antibodies are raised against non-human primate antigens, and databases of cross-reactivity between species are incomplete. Thus, we screened 332 antibodies in five immune cell populations in blood from humans and four non-human primate species generating a comprehensive cross-reactivity catalog that includes cell type-specificity. We used this catalog to create large mass cytometry universal cross-species phenotyping and signaling panels for humans, along with three of the model organisms most similar to humans: rhesus and cynomolgus macaques and African green monkeys; and one of the mammalian models most widely used in drug development: C57BL/6 mice. As a proof-of-principle, we measured immune cell signaling responses across all five species to an array of 15 stimuli using mass cytometry. We found numerous instances of different cellular phenotypes and immune signaling events occurring within and between species, and detailed 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. Antibody panels and the full dataset generated are available online as a resource to enable future studies comparing immune responses across species during the evaluation of therapeutics.

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Section: Resultssupporting

confidence: 84%

A Comprehensive Atlas of Immunological Differences Between Humans, Mice, and Non-Human Primates

et al. 2022

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“…Singlets (DNA mid ) B Cell Distribution (IgM vs CD1c) CD7+ NK Cells broad distribution of CD45RA staining, as previously reported (24,25), with two clones (5H9 and HI100), which was difficult to gate (see online dataset). Staining of this marker was superior with the Versa-Lyse-based method than with the Triton-X100-based method (see methods) (data not shown).…”

Section: Cytof Panel Designmentioning

confidence: 52%

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

Bjornson-Hooper

Fragiadakis

Spitzer

et al. 2019

Preprint

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IntroductionNon-human primates (NHPs) are critical components of drug development because of their similarity to humans. Many key immunology assays, such as flow cytometry, Western blots, immunohistochemistry and immunofluorescence microscopy, make use of antibodies to demarcate specific cell types and quantify signaling moieties. Very few antibodies are raised against non-human primate antigens; instead, researchers typically use anti-human antibodies that are cross-reactive with the non-human primate species that they are studying. To help researchers find antibodies for NHP research, the National Institutes of Health supports a highly valuable database of the cross-reactivity of commercially available antibodies with 13 NHP species (http://www.nhpreagents.org). The database is derived from manufacturer and investigator reports, and typically provides a simple yes/no statement about whether a clone stains a species, with occasional comments about staining intensity or specificity. While an invaluable resource, the database is limited in its coverage. For example, prior to this study, only 28 CD markers had been evaluated in African green monkeys.Additionally, with few exceptions, the database lacks information about the cell types bound by cross-reactive antibodies, and there are many known instances of antibody clones binding different cell types in different species. For example, granulocyte and monocyte marker expression is known to be substantially different in humans than in non-human primates. Anti-human CD33 clone AC104.3E3 was reported in the NIH database and manufacturer's datasheet as cross-reactive with rhesus and cynomolgus macaques, but our lab and others determined that in those species, it prominently stains granulocytes (1, 2), while in humans it stains monocytes and classical dendritic cells. As another example, the Fcγ receptor CD16 is found on granulocytes in humans and sooty mangabeys, but not in macaques or baboons (3, 4), which will likely confound animal studies evaluating therapeutic antibodies, which may bind, transduce signals through and mediate internalization via this Fcγ receptor. Yet another example is CD56, which is expressed on monocytes in macaques (5), but is a canonical NK cell marker in humans. Thus, researchers must confirm that each clone they use is staining the cell population of interest through literature review or experimental verification.Here we present an expansion of both the breadth and depth of primate cross-reactivity data. We screened 332 monoclonal antibodies in blood from two individuals of each of four NHP species: rhesus macaque (Macaca mulatta), cynomolgus macaque (Macaca fascicularis), African green monkey (Chlorcebus aethiops) and olive/ yellow baboon (Papio hamadryas anubis x Papio hamadryas cynocephalus hybrid); and found more than 120 clones that stained one or more populations in each species. Furthermore, we included counter-stain antibodies that allowed us to determine staining specificity in at least five major immune cell populations. Data from ...

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“…Animal studies at Advanced BioScience Laboratories (OLAW A3467-01) were performed under IACUC-approved protocol AUP 315. Details of the animal husbandry used at each site have been published previously (47).…”

Section: Methodsmentioning

confidence: 99%

Design andIn VivoCharacterization of Immunoconjugates Targeting HIV gp160

Pincus

Song

Maresh

et al. 2017

J Virol

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The envelope (Env) glycoprotein of HIV is expressed on the surface of productively infected cells and can be used as a target for cytotoxic immunoconjugates (ICs), in which cell-killing moieties, including toxins, drugs, or radionuclides, are chemically or genetically linked to monoclonal antibodies (MAbs) or other targeting ligands. Such ICs could be used to eliminate persistent reservoirs of HIV infection. We have found that MAbs which bind to the external loop of gp41, e.g., MAb 7B2, make highly effective ICs, particularly when used in combination with soluble CD4. We evaluated the toxicity, immunogenicity, and efficacy of the ICs targeted with 7B2 in mice and in simian-human immunodeficiency virus-infected macaques. In the macaques, we tested immunotoxins (ITs), consisting of protein toxins bound to the targeting agent. ITs were well tolerated and initially efficacious but were ultimately limited by their immunogenicity. In an effort to decrease immunogenicity, we tested different toxic moieties, including recombinant toxins, cytotoxic drugs, and tubulin inhibitors. ICs containing deglycosylated ricin A chain prepared from ricin toxin extracted from castor beans were the most effective in killing HIVinfected cells. Having identified immunogenicity as a major concern, we show that conjugation of IT to polyethylene glycol limits immunogenicity. These studies demonstrate that cytotoxic ICs can target virus-infected cells in vivo but also highlight potential problems to be addressed. IMPORTANCE It is not yet possible to cure HIV infection. Even after years of fully effective antiviral therapy, a persistent reservoir of virus-infected cells remains. Here we propose that a targeted conjugate consisting of an anti-HIV antibody bound to a toxic moiety could function to kill the HIV-infected cells that constitute this reservoir. We tested this approach in HIV-infected cells grown in the lab and in animal infections. Our studies demonstrated that these immunoconjugates are effective both in vitro and in test animals. In particular, ITs constructed with the deglycosylated A chain prepared from native ricin were the most effective in killing cells, but their utility was blunted because they provoked immune reactions that interfered with the therapeutic effects. We then demonstrated that coating of the ITs with polyethylene glycol minimized the immunogenicity, as has been demonstrated with other protein therapies.

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Expression of the Memory Marker CD45RO on Helper T Cells in Macaques

Cited by 11 publications

References 37 publications

A Comprehensive Atlas of Immunological Differences Between Humans, Mice, and Non-Human Primates

A Comprehensive Atlas of Immunological Differences Between Humans, Mice, and Non-Human Primates

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

Design andIn VivoCharacterization of Immunoconjugates Targeting HIV gp160

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