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  1. Article ; Online: Reply to "Risk of severe COVID-19 infection in International Space Station astronauts despite routine pre-mission measures".

    Crucian, Brian E / Makedonas, George / Mehta, Satish K / Haddon, Robert / Scheuring, Richard A

    The journal of allergy and clinical immunology. In practice

    2021  Volume 9, Issue 9, Page(s) 3527–3528

    MeSH term(s) Astronauts ; COVID-19 ; Humans ; SARS-CoV-2 ; Space Flight ; Time Factors
    Language English
    Publishing date 2021-09-08
    Publishing country United States
    Document type Letter ; Comment
    ZDB-ID 2843237-X
    ISSN 2213-2201 ; 2213-2198
    ISSN (online) 2213-2201
    ISSN 2213-2198
    DOI 10.1016/j.jaip.2021.06.017
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Attenuation of GARP expression on regulatory T cells by protein transport inhibitors.

    Anvari, Sara / Schuster, Kimberly / Grimbergen, Andrea / Davis, Carla M / Makedonas, George

    Journal of immunological methods

    2021  Volume 492, Page(s) 112998

    Abstract: An integrated understanding of the functional capacities of cells in the context of their physical parameters and molecular markers is increasingly demanded in immunologic studies. Regulatory T cells (Tregs) are a subpopulation of T cells involved in ... ...

    Abstract An integrated understanding of the functional capacities of cells in the context of their physical parameters and molecular markers is increasingly demanded in immunologic studies. Regulatory T cells (Tregs) are a subpopulation of T cells involved in immune response modulation and mediating tolerance to self-antigen with their absence leading to a loss of tolerance. Glycoprotein repetitions A predominant (GARP) is a key marker for activated Tregs, but its detection may also be useful in determining the functional capacities of the cell. This study aims to deduce the optimal stimulation period and the impact of protein transport inhibitors (PTIs), commonly used in the detection of intracellular cytokines, on GARP detection. Through flow cytometric analysis we analyzed different cell culture conditions for optimal GARP expression on activated Tregs. Healthy donor PBMCs were stimulated with either Staphylococcal Enterotoxin B (SEB) or PMA/Ionomycin (PMA/Iono), in the presence and absence of PTIs monensin and/or brefeldin A (BFA) and GARP expression was assessed on CD4+ CD25+ FOXP3+ Tregs. The optimal stimulation period for the detection of GARP was highest at 24-h. Furthermore, we determined that GARP expression on Tregs is significantly reduced when cells are treated with the PTIs monensin and/or BFA following PMA/Iono stimulation. This effect was not seen following SEB stimulation. Therefore, due to the effects of PTIs, alternative methods should be considered when performing simultaneous analysis for cytokine expression and GARP expression on Tregs.
    MeSH term(s) Brefeldin A/pharmacology ; Cells, Cultured ; Cytokines/analysis ; Cytokines/metabolism ; Enterotoxins/immunology ; Healthy Volunteers ; Humans ; Immunophenotyping/methods ; Lymphocyte Activation ; Membrane Proteins/analysis ; Membrane Proteins/antagonists & inhibitors ; Membrane Proteins/metabolism ; Monensin/pharmacology ; Primary Cell Culture ; T-Lymphocytes, Regulatory/drug effects ; T-Lymphocytes, Regulatory/immunology ; T-Lymphocytes, Regulatory/metabolism
    Chemical Substances Cytokines ; Enterotoxins ; LRRC32 protein, human ; Membrane Proteins ; Brefeldin A (20350-15-6) ; enterotoxin B, staphylococcal (39424-53-8) ; Monensin (906O0YJ6ZP)
    Language English
    Publishing date 2021-02-15
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 120142-6
    ISSN 1872-7905 ; 0022-1759
    ISSN (online) 1872-7905
    ISSN 0022-1759
    DOI 10.1016/j.jim.2021.112998
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: SARS-CoV-2 Pandemic Impacts on NASA Ground Operations to Protect ISS Astronauts.

    Makedonas, George / Mehta, Satish K / Scheuring, Richard A / Haddon, Robert / Crucian, Brian E

    The journal of allergy and clinical immunology. In practice

    2020  Volume 8, Issue 10, Page(s) 3247–3250

    Abstract: NASA implements required medical tests and clinical monitoring to ensure the health and safety of its astronauts. These measures include a pre-launch quarantine to mitigate the risk of infectious diseases. During space missions, most astronauts ... ...

    Abstract NASA implements required medical tests and clinical monitoring to ensure the health and safety of its astronauts. These measures include a pre-launch quarantine to mitigate the risk of infectious diseases. During space missions, most astronauts experience perturbations to their immune system that manifest as a detectable secondary immunodeficiency. On return to Earth, after the stress of re-entry and landing, astronauts would be most vulnerable to infectious disease. In April 2020, a crew returned from International Space Station to NASA Johnson Space Center in Houston, Texas, during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. Post-flight quarantine protocols (both crew and contacts) were enhanced to protect this crew from SARS-CoV-2. In addition, specific additional clinical monitoring was performed to determine post-flight immunocompetence. Given that coronavirus disease 2019 (COVID-19) prognosis is more severe for the immunocompromised, a countermeasures protocol for spaceflight suggested by an international team of scientists could benefit terrestrial patients with secondary immunodeficiency.
    MeSH term(s) Astronauts ; Betacoronavirus ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/immunology ; COVID-19 ; Coronavirus Infections/immunology ; Coronavirus Infections/prevention & control ; Dietary Supplements ; Exercise Therapy ; Granulocyte Colony-Stimulating Factor/therapeutic use ; Humans ; Immunocompromised Host/immunology ; Immunoglobulin G/therapeutic use ; Interleukin-2/therapeutic use ; Organizational Policy ; Pandemics/prevention & control ; Pneumonia, Viral/immunology ; Pneumonia, Viral/prevention & control ; Quarantine/methods ; Quarantine/organization & administration ; SARS-CoV-2 ; Space Flight ; Spacecraft ; Stress, Physiological/immunology ; Texas ; United States ; United States National Aeronautics and Space Administration
    Chemical Substances Immunoglobulin G ; Interleukin-2 ; Granulocyte Colony-Stimulating Factor (143011-72-7)
    Keywords covid19
    Language English
    Publishing date 2020-09-21
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2843237-X
    ISSN 2213-2201 ; 2213-2198
    ISSN (online) 2213-2201
    ISSN 2213-2198
    DOI 10.1016/j.jaip.2020.08.064
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Recycling endosomes in human cytotoxic T lymphocytes constitute an auxiliary intracellular trafficking pathway for newly synthesized perforin.

    Lesteberg, Kelsey / Orange, Jordan / Makedonas, George

    Immunologic research

    2017  Volume 65, Issue 5, Page(s) 1031–1045

    Abstract: Although cytotoxic T lymphocytes (CTLs) store perforin within cytoplasmic secretory granules for immediate use, perforin is synthesized anew within hours of TCR stimulation. Previously, we observed new perforin protein at an immunologic synapse ... ...

    Abstract Although cytotoxic T lymphocytes (CTLs) store perforin within cytoplasmic secretory granules for immediate use, perforin is synthesized anew within hours of TCR stimulation. Previously, we observed new perforin protein at an immunologic synapse independent of secretory lysosomes; herein, we aimed to determine how new perforin transits to the synapse if not via lytic granules. We analyzed antigen-specific human CTLs via imaging flow cytometry and high-resolution confocal microscopy, with attention to intracellular trafficking components and new perforin. The recycling endosome compartments identified by rab8, rab11a, rab4, and rab37 co-localized with new perforin, as well as the SNAREs vti1b and VAMP4. After ablating the function of the recycling endosome pathway, we observed a relative accumulation of new perforin in rab8 vesicles. The recycling endosome pathway may serve as an auxiliary intracellular route for the delivery of new perforin to an immunologic synapse in order to perpetuate a cytotoxic response.
    MeSH term(s) Cells, Cultured ; Cytotoxicity, Immunologic ; Endosomes/metabolism ; Humans ; Immunological Synapses/metabolism ; Perforin/metabolism ; Protein Transport ; Qb-SNARE Proteins/metabolism ; R-SNARE Proteins/metabolism ; Secretory Vesicles/metabolism ; T-Lymphocytes, Cytotoxic/immunology ; rab GTP-Binding Proteins/metabolism
    Chemical Substances Qb-SNARE Proteins ; R-SNARE Proteins ; VAMP4 protein, human ; VTI1B protein, human ; Perforin (126465-35-8) ; rab GTP-Binding Proteins (EC 3.6.5.2)
    Language English
    Publishing date 2017
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 632857-x
    ISSN 1559-0755 ; 0257-277X
    ISSN (online) 1559-0755
    ISSN 0257-277X
    DOI 10.1007/s12026-017-8945-8
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Protein transport inhibitors downregulate the expression of LAG-3 on regulatory T cells.

    Anvari, Sara / Grimbergen, Andrea / Davis, Carla M / Makedonas, George

    Journal of immunological methods

    2017  Volume 447, Page(s) 47–51

    Abstract: Modern immunologic studies demand increasing complexity because of a need to improve our understanding of the relationship between a cell's phenotype and its function. Regulatory T cells (Tregs) have been defined by a narrow set of phenotypic markers, ... ...

    Abstract Modern immunologic studies demand increasing complexity because of a need to improve our understanding of the relationship between a cell's phenotype and its function. Regulatory T cells (Tregs) have been defined by a narrow set of phenotypic markers, however their actual functional capacity has not been determined at the single-cell level. Although the lymphocyte activation gene 3 (LAG-3; CD223) is a key marker for the identification of exhausted T cells, it may be useful also in resolving Treg subpopulations by indicating distinct functional breadths. Here we define the experimental conditions necessary for the optimal detection by flow cytometry of LAG-3 expression on activated Tregs. We stimulated human PBMCs with either PMA/ionomycin or Staphylococcal Enterotoxin B (SEB) and analyzed CD4+CD25+FoxP3+ Tregs for LAG-3 expression in concert with other Treg phenotypic markers. We prescribe a 24-hour stimulation period for the optimal detection of LAG-3 on Tregs. Furthermore, we determine LAG-3 protein expression on Tregs is compromised when the cells are treated with brefeldin A (BFA) and monensin. Therefore, the simultaneous assessment of Treg phenotype and function is complicated by the use of protein transport inhibitors.
    MeSH term(s) Antigens, CD/genetics ; Antigens, CD/immunology ; Biomarkers ; Brefeldin A/pharmacology ; Cells, Cultured ; Down-Regulation ; Enterotoxins/pharmacology ; Flow Cytometry ; Humans ; Immunophenotyping ; Ionomycin/pharmacology ; Leukocytes, Mononuclear/drug effects ; Leukocytes, Mononuclear/immunology ; Lymphocyte Activation ; Monensin/pharmacology ; Protein Transport ; T-Lymphocytes, Regulatory/drug effects ; T-Lymphocytes, Regulatory/immunology
    Chemical Substances Antigens, CD ; Biomarkers ; CD223 antigen ; Enterotoxins ; Brefeldin A (20350-15-6) ; enterotoxin B, staphylococcal (39424-53-8) ; Ionomycin (56092-81-0) ; Monensin (906O0YJ6ZP)
    Language English
    Publishing date 2017-08
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 120142-6
    ISSN 1872-7905 ; 0022-1759
    ISSN (online) 1872-7905
    ISSN 0022-1759
    DOI 10.1016/j.jim.2017.04.006
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: An evaluation of cytokine and cellular immune responses to heterologous prime-boost vaccination with influenza A/H7N7-A/H7N9 inactivated vaccine.

    El Sahly, Hana M / Makedonas, George / Corry, David / Atmar, Robert L / Bellamy, Abbie / Cross, Kaitlyn / Keitel, Wendy A

    Human vaccines & immunotherapeutics

    2020  Volume 16, Issue 12, Page(s) 3138–3145

    Abstract: The immunologic mechanisms underlying the improved serologic responses to heterologous prime-boost avian influenza vaccination are unclear. An exploratory analysis of the immune responses following 1 dose of influenza A/H7N9 inactivated vaccine in ... ...

    Abstract The immunologic mechanisms underlying the improved serologic responses to heterologous prime-boost avian influenza vaccination are unclear. An exploratory analysis of the immune responses following 1 dose of influenza A/H7N9 inactivated vaccine in subjects who received an influenza A/H7N7 inactivated vaccine (N = 17) 8 years earlier or who were influenza A/H7-naïve (10) was performed. Plasma IL-6 and IL-21 concentrations by ELISA, the frequency of A/H7N7-specific memory B cells and antibody secreting cells by ELISpot, the frequency of circulating T follicular helper cells and the frequency of T cells expressing IL-6 and IL-21 by flow cytometry were assessed at baseline (D1), and 8 days (D9) and 28 days (D29) after vaccination. We assessed the correlation between these measurements and the D29 serologic responses to the boost vaccine. Plasma IL-6 concentration on D9 significantly correlated with the H7N7 and H7N9 hemagglutination inhibition (HAI) antibody levels (
    MeSH term(s) Animals ; Antibodies, Viral ; Birds ; Cytokines ; Humans ; Immunity, Cellular ; Influenza A Virus, H7N7 Subtype/immunology ; Influenza A Virus, H7N9 Subtype/immunology ; Influenza Vaccines ; Influenza in Birds ; Influenza, Human/prevention & control ; Vaccination ; Vaccines, Inactivated
    Chemical Substances Antibodies, Viral ; Cytokines ; Influenza Vaccines ; Vaccines, Inactivated
    Language English
    Publishing date 2020-05-13
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2664176-8
    ISSN 2164-554X ; 2164-5515
    ISSN (online) 2164-554X
    ISSN 2164-5515
    DOI 10.1080/21645515.2020.1750910
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Living in a house of cards: re-evaluating CD8+ T-cell immune correlates against HIV.

    Makedonas, George / Betts, Michael R

    Immunological reviews

    2011  Volume 239, Issue 1, Page(s) 109–124

    Abstract: The Merck STEP and the Thai RV144 human immunodeficiency virus (HIV) vaccine trials confirmed that we still have a long way to go before developing a prophylactic HIV vaccine. The main issue at hand is that we have yet to identify an immunological ... ...

    Abstract The Merck STEP and the Thai RV144 human immunodeficiency virus (HIV) vaccine trials confirmed that we still have a long way to go before developing a prophylactic HIV vaccine. The main issue at hand is that we have yet to identify an immunological correlate of protection against HIV. While many question the T-cell-based approach towards vaccine development, it is likely that T cells will be a necessary part of any vaccine strategy. CD8(+) T cells remain an attractive option because of their ability to specifically recognize and eliminate virally infected host cells. In this review, we recapitulate the evidence for CD8(+) T cells as an immunological correlate against HIV, but more importantly, we assess the means by which we evaluate their antiviral capacity. To achieve a breakthrough in the domain of T-cell-based HIV vaccine development, it has become abundantly clear that we must overhaul our system of immune monitoring and come up with a 'rational' tactic to evaluate the efficacy of HIV-specific CD8(+) T cells.
    MeSH term(s) AIDS Vaccines/genetics ; AIDS Vaccines/immunology ; Adaptive Immunity ; CD8-Positive T-Lymphocytes/immunology ; Clinical Trials as Topic ; Cytokines/immunology ; Cytokines/metabolism ; HIV/immunology ; HIV/physiology ; HIV Infections/immunology ; HIV Infections/prevention & control ; Humans ; Lymphocyte Activation ; Perforin/metabolism
    Chemical Substances AIDS Vaccines ; Cytokines ; Perforin (126465-35-8)
    Language English
    Publishing date 2011-01-19
    Publishing country England
    Document type Journal Article ; Review
    ZDB-ID 391796-4
    ISSN 1600-065X ; 0105-2896
    ISSN (online) 1600-065X
    ISSN 0105-2896
    DOI 10.1111/j.1600-065X.2010.00968.x
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article: SARS-CoV-2 Pandemic Impacts on NASA Ground Operations to Protect ISS Astronauts

    Makedonas, George / Mehta, Satish K / Scheuring, Richard A / Haddon, Robert / Crucian, Brian E

    J Allergy Clin Immunol Pract

    Abstract: NASA implements required medical tests and clinical monitoring to ensure the health and safety of its astronauts. These measures include a pre-launch quarantine to mitigate the risk of infectious diseases. During space missions, most astronauts ... ...

    Abstract NASA implements required medical tests and clinical monitoring to ensure the health and safety of its astronauts. These measures include a pre-launch quarantine to mitigate the risk of infectious diseases. During space missions, most astronauts experience perturbations to their immune system that manifest as a detectable secondary immunodeficiency. On return to Earth, after the stress of re-entry and landing, astronauts would be most vulnerable to infectious disease. In April 2020, a crew returned from International Space Station to NASA Johnson Space Center in Houston, Texas, during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. Post-flight quarantine protocols (both crew and contacts) were enhanced to protect this crew from SARS-CoV-2. In addition, specific additional clinical monitoring was performed to determine post-flight immunocompetence. Given that coronavirus disease 2019 (COVID-19) prognosis is more severe for the immunocompromised, a countermeasures protocol for spaceflight suggested by an international team of scientists could benefit terrestrial patients with secondary immunodeficiency.
    Keywords covid19
    Publisher WHO
    Document type Article
    Note WHO #Covidence: #779133
    Database COVID19

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  9. Article ; Online: SARS-CoV-2 Pandemic Impacts on NASA Ground Operations to Protect ISS Astronauts

    Makedonas, George / Mehta, Satish K. / Scheuring, Richard A. / Haddon, Robert / Crucian, Brian E.

    The Journal of Allergy and Clinical Immunology: In Practice

    2020  Volume 8, Issue 10, Page(s) 3247–3250

    Keywords Immunology and Allergy ; covid19
    Language English
    Publisher Elsevier BV
    Publishing country us
    Document type Article ; Online
    ZDB-ID 2843237-X
    ISSN 2213-2201 ; 2213-2198
    ISSN (online) 2213-2201
    ISSN 2213-2198
    DOI 10.1016/j.jaip.2020.08.064
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  10. Article ; Online: Spaceflight validation of technology for point-of-care monitoring of peripheral blood WBC and differential in astronauts during space missions.

    Crucian, Brian / Valentine, Russell / Calaway, Kimesha / Miller, Rachael / Rubins, Kathleen / Hopkins, Michael / Salas, Zachary / Krieger, Stephanie / Makedonas, George / Nelman-Gonzalez, Mayra / McMonigal, Kathleen / Perusek, Gail / Lehnhardt, Kris / Easter, Benjamin

    Life sciences in space research

    2021  Volume 31, Page(s) 29–33

    Abstract: During long duration orbital space missions, astronauts experience immune system dysregulation, the persistent reactivation of latent herpesviruses, and some degree of clinical incidence. During planned NASA 'Artemis' deep space missions the stressors ... ...

    Abstract During long duration orbital space missions, astronauts experience immune system dysregulation, the persistent reactivation of latent herpesviruses, and some degree of clinical incidence. During planned NASA 'Artemis' deep space missions the stressors that cause this phenomenon will increase, while clinical care capability will likely be reduced. There is currently minimal clinical laboratory capability aboard the International Space Station (ISS). The ability to monitor the white blood cell count (WBC) and differential during spaceflight has been an unmet NASA medical requirement, primarily due to a lack of capable hardware. We performed ground and flight validation of a device designed to monitor WBC and differential within minutes from a fingerstick blood sample. This device is miniaturized, robust, and generally compatible with microgravity operations. Ground testing for spaceflight consisted of vibration tolerance, power/battery and interface requirements, electromagnetic interference (EMI), and basic evaluation of sample preparation and operations in the context of spaceflight constraints. The in-flight validation performed aboard the ISS by two astronauts included assessment of three levels of control solution (blood) samples as well as a real time analysis of a fingerstick blood sample by one of the crewmembers. Flight and ground testing of the same lot of control solutions yielded similar total WBC values. There was some select discrepancy between flight and ground data for the differential analysis. However, the data suggest that this issue is due to compromise of the control solutions as a result of storage length before flight operations, and not due to a microgravity-associated issue with instrument performance. This evaluation also yielded lessons learned regarding crewmember training for technique-sensitive small-volume biosample collection and handling in microgravity. The fingerstick analysis was successful and was the first real-time hematology assessment performed during spaceflight. This device may provide an in-mission monitoring capability for astronauts thereby assisting Flight Surgeons and the crew medical officer during both orbital and deep space missions.
    MeSH term(s) Astronauts ; Humans ; Point-of-Care Systems ; Space Flight ; Technology ; Weightlessness/adverse effects
    Language English
    Publishing date 2021-07-15
    Publishing country Netherlands
    Document type Journal Article
    ISSN 2214-5532
    ISSN (online) 2214-5532
    DOI 10.1016/j.lssr.2021.07.003
    Database MEDical Literature Analysis and Retrieval System OnLINE

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