Resource and Technical AdvanceIn-Press PreviewImmunologyInfectious disease
Open Access | 10.1172/jci.insight.185861
1Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States of America
2Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, United States of America
3Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, United States of America
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1Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States of America
2Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, United States of America
3Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, United States of America
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1Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States of America
2Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, United States of America
3Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, United States of America
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1Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States of America
2Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, United States of America
3Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, United States of America
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1Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States of America
2Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, United States of America
3Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, United States of America
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1Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States of America
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3Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, United States of America
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1Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States of America
2Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, United States of America
3Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, United States of America
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1Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States of America
2Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, United States of America
3Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, United States of America
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1Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States of America
2Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, United States of America
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1Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States of America
2Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, United States of America
3Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, United States of America
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Published April 17, 2025 - More info
We developed a 29-color spectral cytometry panel to enhance nonhuman primate (NHP) models for cross-reactive immunophenotyping. This panel is suitable for biosafety level 4 (BSL-4) viruses and can be used with both human and NHP samples in BSL-2 research settings. Tissues from humans, rhesus monkeys (RhMs), crab-eating macaques (CEMs), and green monkeys (GMs) were stained with a 29-color immunophenotyping panel requiring only two clone substitutions. Comparable staining was observed for all samples. Unbiased analysis showed acceptable overlap in T-cell phenotypes across samples, with differences in human and NHP B cells and granulocytes. In CEMs, most circulating CD8+ T cells were from effector memory cells, with significantly higher levels than in humans (p<0.0001), RhMs (p<0.05), and GMs (p<0.01). Analysis of samples from various anatomical sites revealed distinct location-specific phenotypes. In Nipah-virus-exposed animals, splenocytes showed a substantial increase in IgM+ B cells (p<0.0001) and a reduction in effector memory CD8+ T cells (p<0.0001) compared to unexposed controls. Lymph nodes from Ebola-virus-exposed animals showed a loss of CXCR3+CD8+ T cells vs unexposed controls. This panel may guide the development of additional multi-color panels in preclinical and clinical settings and potentially increase understanding of the pathogenesis of diseases caused by emerging and re-emerging viruses.