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Статті в журналах з теми "Blood cell imaging"

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Автор: Grafiati

Опубліковано: 6 вересня 2023

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1

GUZE, BARRY H., RANDALL A. HAWKINS, and CAROL S. MARCUS. "Technetium-99m White Blood Cell Imaging." Clinical Nuclear Medicine 14, no. 2 (February 1989): 104–6. http://dx.doi.org/10.1097/00003072-198902000-00007.

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2

Tishko, Tatyana V., Dimitrij Tishko, and Vladimir Titar. "Holographic Method for Blood Cell Imaging." Imaging & Microscopy 11, no. 3 (August 2009): 46–48. http://dx.doi.org/10.1002/imic.200990063.

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3

KURTULDU, Hüseyin, Aynur Didem OKTAN, Hatice CANDAN, and Beste Sahra CİHANGİROĞLU. "Red Blood Cell Analysis by Hyperspectral Imaging." Natural and Applied Sciences Journal 1, no. 2 (December 29, 2018): 1–7. http://dx.doi.org/10.38061/idunas.442490.

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4

Mittelbrunn, María, Gloria Martínez del Hoyo, María López-Bravo, Noa B. Martín-Cofreces, Alix Scholer, Stéphanie Hugues, Luc Fetler, Sebastián Amigorena, Carlos Ardavín, and Francisco Sánchez-Madrid. "Imaging of plasmacytoid dendritic cell interactions with T cells." Blood 113, no. 1 (January 1, 2009): 75–84. http://dx.doi.org/10.1182/blood-2008-02-139865.

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Abstract Plasmacytoid dendritic cells (pDCs) efficiently produce type I interferon and participate in adaptive immune responses, although the molecular interactions between pDCs and antigen-specific T cells remain unknown. This study examines immune synapse (IS) formation between murine pDCs and CD4+ T cells. Mature pDCs formed canonical ISs, involving relocation to the contact site of the microtubule-organizing center, F-actin, protein kinase C-θ, and pVav, and activation of early signaling molecules in T cells. However, immature pDCs were less efficient at forming conjugates with T cells and inducing IS formation, microtubule-organizing center translocation, and T-cell signaling and activation. Time-lapse videomicroscopy and 2-photon in vivo imaging of pDC–T-cell interactions revealed that immature pDCs preferentially mediated transient interactions, whereas mature pDCs promoted more stable contacts. Our data indicate that, under steady-state conditions, pDCs preferentially establish transient contacts with naive T cells and show a very modest immunogenic capability, whereas on maturation, pDCs are able to form long-lived contacts with T cells and significantly enhance their capacity to activate these lymphocytes.

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5

Loeffler, Dirk, and Timm Schroeder. "Understanding cell fate control by continuous single-cell quantification." Blood 133, no. 13 (March 28, 2019): 1406–14. http://dx.doi.org/10.1182/blood-2018-09-835397.

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Abstract Cells and the molecular processes underlying their behavior are highly dynamic. Understanding these dynamic biological processes requires noninvasive continuous quantitative single-cell observations, instead of population-based average or single-cell snapshot analysis. Ideally, single-cell dynamics are measured long-term in vivo; however, despite progress in recent years, technical limitations still prevent such studies. On the other hand, in vitro studies have proven to be useful for answering long-standing questions. Although technically still demanding, long-term single-cell imaging and tracking in vitro have become valuable tools to elucidate dynamic molecular processes and mechanisms, especially in rare and heterogeneous populations. Here, we review how continuous quantitative single-cell imaging of hematopoietic cells has been used to solve decades-long controversies. Because aberrant cell fate decisions are at the heart of tissue degeneration and disease, we argue that studying their molecular dynamics using quantitative single-cell imaging will also improve our understanding of these processes and lead to new strategies for therapies.

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6

Zhang, Yi-Yi, Jia-Chen Wu, Ran Hao, Shang-Zhong Jin, and Liang-Cai Cao. "Digital holographic microscopy for red blood cell imaging." Acta Physica Sinica 69, no. 16 (2020): 164201. http://dx.doi.org/10.7498/aps.69.20200357.

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7

Hanssen, E., C. Knoechel, P. Carlton, J. Sedat, C. Larabell, and L. Tilley. "Whole Cell Imaging of Plasmodium Falciparum Blood Stages." Microscopy and Microanalysis 15, S2 (July 2009): 866–67. http://dx.doi.org/10.1017/s143192760909268x.

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8

Debatin, J�rg F., and Erol M. Beytas. "Indium 111 white blood cell imaging of epididymitis." European Journal of Nuclear Medicine 17, no. 5 (1990): 286–89. http://dx.doi.org/10.1007/bf00812372.

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9

Al-Janabi, M. A., P. J. Maltby, M. Critchley, and K. E. Britton. "5. Radiolabelled white blood cell imaging. Is it a blood pool effect?" Nuclear Medicine Communications 11, no. 12 (December 1990): 890. http://dx.doi.org/10.1097/00006231-199012000-00011.

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10

Vynckier, Jan, Jelle Demeestere, and Julie Lambert. "Black-blood Magnetic Resonance Imaging in Giant Cell Arteritis." Journal of Rheumatology 48, no. 2 (February 2021): 301–2. http://dx.doi.org/10.3899/jrheum.190286.

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11

CHANDRAMOULY, B., T. SCAGNELL, P. NARDI, and A. BRUDNICKI. "lndium-111 White Blood Cell Imaging in Suppurative Peritonitis." Clinical Nuclear Medicine 15, no. 4 (April 1990): 268–69. http://dx.doi.org/10.1097/00003072-199004000-00016.

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12

Popescu, Gabriel, YoungKeun Park, Wonshik Choi, Ramachandra R. Dasari, Michael S. Feld, and Kamran Badizadegan. "Imaging red blood cell dynamics by quantitative phase microscopy." Blood Cells, Molecules, and Diseases 41, no. 1 (July 2008): 10–16. http://dx.doi.org/10.1016/j.bcmd.2008.01.010.

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13

Winkelman, James W. "Noninvasive Blood Cell Measurements by Imaging of the Microcirculation." American Journal of Clinical Pathology 113, no. 4 (April 1, 2000): 479–83. http://dx.doi.org/10.1309/7079-v61f-d90u-gu6r.

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14

Cortes, Julia, Juan I. Alonso, Francisco Ruiz–Oliva, Soledad Alvarez, Julio Saenz Ormijana, Blanca Caton, and Pilar Alcorta. "Renal Cell Carcinoma Detected on Tc-99m-Labeled Red Blood Cell Imaging." Clinical Nuclear Medicine 28, no. 11 (November 2003): 920–22. http://dx.doi.org/10.1097/01.rlu.0000093089.53875.3e.

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15

Yi, Faliu, Inkyu Moon, and Bahram Javidi. "Cell morphology-based classification of red blood cells using holographic imaging informatics." Biomedical Optics Express 7, no. 6 (May 25, 2016): 2385. http://dx.doi.org/10.1364/boe.7.002385.

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16

AL-JANABI, M. A., M. CRITCHLEY, P. MALTBY, and K. E. BRITTON. "Radiolabelled white blood cell imaging in arthritis. Is it a blood pool effect?" Nuclear Medicine Communications 12, no. 12 (December 1991): 1013–24. http://dx.doi.org/10.1097/00006231-199112000-00003.

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17

Severo, Maiara S., Jonathan J. M. Landry, Randall L. Lindquist, Christian Goosmann, Volker Brinkmann, Paul Collier, Anja E. Hauser, et al. "Unbiased classification of mosquito blood cells by single-cell genomics and high-content imaging." Proceedings of the National Academy of Sciences 115, no. 32 (July 23, 2018): E7568—E7577. http://dx.doi.org/10.1073/pnas.1803062115.

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Анотація:

Mosquito blood cells are immune cells that help control infection by vector-borne pathogens. Despite their importance, little is known about mosquito blood cell biology beyond morphological and functional criteria used for their classification. Here, we combined the power of single-cell RNA sequencing, high-content imaging flow cytometry, and single-molecule RNA hybridization to analyze a subset of blood cells of the malaria mosquito Anopheles gambiae. By demonstrating that blood cells express nearly half of the mosquito transcriptome, our dataset represents an unprecedented view into their transcriptional program. Analyses of differentially expressed genes identified transcriptional signatures of two cell types and provide insights into the current classification of these cells. We further demonstrate the active transfer of a cellular marker between blood cells that may confound their identification. We propose that cell-to-cell exchange may contribute to cellular diversity and functional plasticity seen across biological systems.

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18

Maeda, Yoshiaki, Tomoko Yoshino, Atsushi Kogiso, Ryo Negishi, Tomohiro Takabayashi, Hikaru Tago, Tae-Kyu Lim, Manabu Harada, Tadashi Matsunaga, and Tsuyoshi Tanaka. "Lensless imaging-based discrimination between tumour cells and blood cells towards circulating tumour cell cultivation." Analyst 146, no. 23 (2021): 7327–35. http://dx.doi.org/10.1039/d1an01414e.

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19

Jaferzadeh, Keyvan, Inkyu Moon, Manon Bardyn, Michel Prudent, Jean-Daniel Tissot, Benjamin Rappaz, Bahram Javidi, Gerardo Turcatti, and Pierre Marquet. "Quantification of stored red blood cell fluctuations by time-lapse holographic cell imaging." Biomedical Optics Express 9, no. 10 (September 10, 2018): 4714. http://dx.doi.org/10.1364/boe.9.004714.

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20

Liu, Xiaoshuai, Yuchao Li, Xiaohao Xu, Yao Zhang, and Baojun Li. "Red-Blood-Cell-Based Microlens: Application to Single-Cell Membrane Imaging and Stretching." ACS Applied Bio Materials 2, no. 7 (May 16, 2019): 2889–95. http://dx.doi.org/10.1021/acsabm.9b00274.

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21

Acevedo-Mendez, Maria, Anil Rao, and Lewis L. Hsu. "Diagnostic Imaging Radiation in Severe Sickle Cell Disease: Cancer Risk Implications." Blood 138, Supplement 1 (November 5, 2021): 4944. http://dx.doi.org/10.1182/blood-2021-149911.

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Abstract Introduction: Patients with sickle cell disease (SCD) are repeatedly exposed to diagnostic radiation. Radiographs, computed tomography (CT) and nuclear medicine scans are often ordered for suspected complications caused by sickle cell disease that exposes patients with SCD to ionizing or another form of radiation. A few studies of low-dose cumulative radiation exposure (in people without SCD) suggest that 30 to 100 mSv over 30 decades is associated with higher excess risk of leukemia. New epidemiologic data of low quality suggests that individuals with sickle cell disease (SCD) accumulate "driver mutations" for acute myelogenous leukemia (AML) about 20-30 years earlier than the general population, and have higher risk of AML. In a gene therapy protocol with a few dozen patients, 2 cases of AML have occurred in sickle cell disease and none in thalassemia. It has been reported that children with SCD are frequently exposed to ionizing radiation in the form of plain radiographs, fluoroscopy, computed tomography (CT) scans, bone scans, and other tests. Exposure to ionizing radiation during childhood carries a risk of developing cancer that is directly related to the total radiation dose.Epidemiological data has demonstrated an increase both in diagnostic radiation and in actual or predicted resultant cancer diagnosis. Children are particularly vulnerable to radiation-induced cancer because they are still actively growing and thus are at greater risk of acquiring an oncogenic mutation in an actively dividing cell. Hypothesis: Frequent diagnostic imaging for children and adults with SCD can have significant cumulative radiation exposure that could add excess risk of AML. Methods: The study design was a retrospective chart review. The sample was selected to be enriched for the most severely-affected children and adults in the Sickle Cell Center at UI Health, which provides medical care for over 700 patients with SCD. The subgroup on chronic erythrocytapheresis blood transfusionswere selected as a sample of severe of SCD who are more likely to be exposed to repeated diagnostic radiation. Many have had stroke, which often leads to repeated head CT and cerebral angiograms. Others had pulmonary embolism or acute chest syndrome, which can lead to chest CT angiograms. Other SCD complications with high risk of morbidity or morbidity lead to similar likelihood that patients on chronic exchange transfusion therapy would have histories of multiple imaging studies. Medical records were reviewed for the type and number of all radiographic tests, especially CT scans, during the 10-year period 2011-2020. A second observer confirmed a subset of charts. Standard references were used to estimate radiation exposure in mSv for each type of test. The sum of mSv for each individual was a rough estimate of cumulative radiation in 10 years. The IRB approved the protocol. Results: Chart review on 39 patients (ages 16 - 60y) identified 1,030 radiographic tests with a mean of 26.4 tests/patient. Seven patients had > 50 tests, and one patient had > 100 tests over a 10-year period. Thirty-three patients had at least one CT scan. Eighteen patients had at least 3 CT scans. Twenty patients had cumulative radiation exposure > 30 mSv over a 10-year period, 4 patients had > 100 mSv, and one patient had > 200 mSv. Plain radiographs comprised 71% (736) of the studies and relatively low dose radiation exposure. Discussion: This retrospective study estimated that diagnostic radiography exposed 20 of 39 patients with severe SCD to the range of 30 to 200 mSv over 10 years, mostly from numerous CT scans. This range of cumulative radiation exposure has mixed evidence about possible heightened risk of AML and other cancers. The study is limited by the small sample at a single institution and a heavy bias toward patients with stroke and chest complications, but this severely-affected subgroup comprises many of those eligible for transplant and gene therapy in SCD. Cumulative exposure to diagnostic radiation might be one mechanism for the unexplained patterns of AML in SCD after gene therapy that led to a pause in SCD gene therapy studies for a few months in 2021. Further studies are needed. Disclosures Hsu: Global Blood Therapeutics: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Aruvant: Consultancy, Membership on an entity's Board of Directors or advisory committees; Hoffman LaRoche: Consultancy, Membership on an entity's Board of Directors or advisory committees; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Forma Therapeutics: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Cyclerion: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Imara: Research Funding; Eli Lilly: Research Funding; Baxalta / Shire / Takeda: Research Funding.

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22

Finkelstein, David M., Arnold M. Noyek, and Joel C. Kirsh. "Red Blood Cell Scan in Cavernous Hemangioma of the Larynx." Annals of Otology, Rhinology & Laryngology 98, no. 9 (September 1989): 707–12. http://dx.doi.org/10.1177/000348948909800909.

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Cavernous hemangioma of the larynx is an uncommon, difficult-to-diagnose vascular tumor for which there is no significant imaging literature to date. The possibility of improved diagnosis through RBC scanning might obviate injudicious biopsy and potential hemorrhage within the airway. Utilizing the radionuclide RBC scan, which labels the patient's own RBCs initially with cold pyrophosphate, and subsequently with technetium 99m as pertechnetate, we have identified successfully four patients with cavernous hemangioma of the larynx. All presented with a supraglottic mass involving at least the aryepiglottic fold and arytenoid region unilaterally. This report describes our satisfactory diagnostic imaging experience with the radionuclide RBC scan and suggests both its imaging specificity and its role in the management of this lesion.

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23

McKay, Gregory N., Ryan C. Niemeier, Carlos Castro-González, and Nicholas J. Durr. "Scattering oblique plane microscopy for in-vivo blood cell imaging." Biomedical Optics Express 12, no. 5 (April 5, 2021): 2575. http://dx.doi.org/10.1364/boe.422993.

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24

Li, Qingli, Mei Zhou, Hongying Liu, Yiting Wang, and Fangmin Guo. "Red Blood Cell Count Automation Using Microscopic Hyperspectral Imaging Technology." Applied Spectroscopy 69, no. 12 (December 2015): 1372–80. http://dx.doi.org/10.1366/14-07766.

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25

Eilken, Hanna M., Shin-Ichi Nishikawa, and Timm Schroeder. "Continuous single-cell imaging of blood generation from haemogenic endothelium." Nature 457, no. 7231 (February 2009): 896–900. http://dx.doi.org/10.1038/nature07760.

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26

Türkmen, Cüneyt, Seher Unal, Yasemin Sanli, and Rejin Kebudi. "Technetium-99m red blood cell imaging of multicentric kaposiform haemangioendothelioma." European Journal of Nuclear Medicine and Molecular Imaging 33, no. 3 (February 17, 2006): 391. http://dx.doi.org/10.1007/s00259-006-1989-y.

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27

Cottereau, Anne-Ségolène. "PET imaging: back in the game for gastric EMZL?" Blood 139, no. 2 (January 13, 2022): 154–55. http://dx.doi.org/10.1182/blood.2021013964.

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28

Herhaus, Peter, Jana Lipkova, Felicitas Lammer, Julia Slotta-Huspenina, Benedikt Wiestler, Tibor Vag, Stefan Habringer, et al. "CXCR4-Targeted Positron Emission Tomography Imaging of Central Nervous System B-Cell Lymphoma." Blood 134, Supplement_1 (November 13, 2019): 2900. http://dx.doi.org/10.1182/blood-2019-126024.

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Introduction Central nervous system lymphoma (CNSL) is a rare neoplasia and arises as primary (PCNSL) or secondary CNSL (SCNSL) and most commonly occurs as diffuse large B-cell lymphoma. Although prognosis of this disease has significantly improved due to advantages in diagnostic procedures and intensification of treatment over the last decade there remain major challenges in the clinical management. Magnetic resonance imaging (MRI) - as the standard imaging modality - has difficulties to discriminate CNSL from other brain-derived tumors or metastasis. Moreover, prognostic scores in CNSL lack the ability to reliable define patients with high relapse risk 1. Therefore, novel strategies to facilitate diagnosis and to select patients that profit from intense treatment protocols are urgently needed. The C-X-C chemokine receptor 4 (CXCR4) is a transmembrane chemokine receptor with pivotal roles in cell homing and is often overexpressed in hematologic malignancies. CXCR4-directed positron emission tomography (PET) imaging with the tracer [68Ga]Pentixafor has been proven to be a suitable in vivo imaging modality for CXCR4 expression in lymphoid malignancies 2. To evaluate the feasibility of CXCR4-directed PET and the prognostic value of this imaging modality this retrospective proof-of-concept study evaluated [68Ga]Pentixafor PET imaging in patients with CNSL. Methods 11 patients with lymphoma of the CNS (n=8 PCNSL, n=3 SCNSL) were imaged with the CXCR4-directed PET tracer [68Ga]Pentixafor in this retrospective proof-of-concept study after signing inform consent. Lymphoma tissue was assessed for CXCR4 expression ex vivo by immunohistochemistry. The prognostic value of CXCR4-directed PET imaging was evaluated in a computed analysis in 7 patients with follow-up MRI. Treatment response calculated as treatment efficiency η by sequential MRI was correlated with [68Ga]Pentixafor PET derived parameters at diagnosis such as volume of PET positive lymphoma lesion V(PET), maximal PET uptake value within the lesion max(PET) or integrated uptake values over the lymphoma lesion ∫(PET). Analysis was performed in a lesion- and patient-based manner. Results [68Ga]Pentixafor PET imaging was positive in all patients with active disease (10/11 patients) with excellent contrast characteristics to the surrounding brain parenchyma. PET positive lesions correlated well with lymphoma lesions in MRI-T1c sequences. The surrounding edema depicted in MRI-FLAIR sequences was evaluated as PET negative (Figure 1). Semi-quantitative analysis revealed maximum standard uptake values of [68Ga]Pentixafor-derived PET from 4.2 to 23.3 within the lesions with a high tumor-to-background-ratio ranging from 13.2 to 83.0. The computed analysis revealed that CXCR4-directed PET parameters at diagnosis given by max(PET) and ∫(PET) significantly correlated with treatment response in the lesion-based as well as in the patient-based analysis. Specifically, ∫(PET) was the most significant prognostic factor in the present study (Figure 2 C, D, F). On the other hand tumor volume at diagnosis measured either by MRI or by [68Ga]Pentixafor PET did not denote a predictive marker for treatment response (Figure 2 A, B, E). Conclusion/Outlook CXCR4-directed PET imaging represents a novel imaging modality for CNSL. Due to its excellent contrast properties it might help to facilitate diagnosis and refine response assessment. Moreover, owing the predictive value for treatment response, CXCR4-directed PET could serve as a biomarker for the selection of patients profiting from intense treatment protocols. Furthermore, the feasibility of endoradiotherapeutic approaches targeting CXCR4 with Pentixather - the therapeutic twin of the imaging peptide Pentixafor - has been shown in hematologic malignancies and could be incorporated in the treatment of CNSL. References 1. Han CH, Batchelor TT. Diagnosis and management of primary central nervous system lymphoma. Cancer. 2017;123(22):4314-4324. 2.Kircher M, Herhaus P, Schottelius M, et al. CXCR4-directed theranostics in oncology and inflammation. Ann Nucl Med. 2018;32(8):503-511. Disclosures Wester: Scintomics: Other: Spouse CEO of Company; CXCR4-targeted radiopharmaceuticals: Other: Inventor; Scintomics GmbH, Germany: Other: Shareholder. Bassermann:Celgene: Consultancy, Research Funding.

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29

T Zalizam, T. Muda, Abdul Salam Rosalina, and Ismail Suzilah. "Adaptive Hybrid Blood Cell Image Segmentation." MATEC Web of Conferences 255 (2019): 01001. http://dx.doi.org/10.1051/matecconf/201925501001.

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Image segmentation is an important phase in the image recognition system. In medical imaging such as blood cell analysis, it becomes a crucial step in quantitative cytophotometry. Currently, blood cell images become predominantly valuable in medical diagnostics tools. In this paper, we present an adaptive hybrid analysis based on selected segmentation algorithms. Three designates common approaches, that are Fuzzy c-means, K-means and Mean-shift are adapted. Blood cell images that are infected with malaria parasites at various stages were tested. The most suitable method will be selected based on the lowest number of regions. The selected approach will be enhanced by applying Median-cut algorithm to further expand the segmentation process. The proposed adaptive hybrid method has shown a significant improvement in the number of regions.

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30

Assmann, Julian C., Don E. Farthing, Keita Saito, Natella Maglakelidze, Brittany Oliver, Kathrynne A. Warrick, Carole Sourbier, et al. "Glycolytic metabolism of pathogenic T cells enables early detection of GVHD by 13C-MRI." Blood 137, no. 1 (January 7, 2021): 126–37. http://dx.doi.org/10.1182/blood.2020005770.

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Abstract Graft-versus-host disease (GVHD) is a prominent barrier to allogeneic hematopoietic stem cell transplantation (AHSCT). Definitive diagnosis of GVHD is invasive, and biopsies of involved tissues pose a high risk of bleeding and infection. T cells are central to GVHD pathogenesis, and our previous studies in a chronic GVHD mouse model showed that alloreactive CD4+ T cells traffic to the target organs ahead of overt symptoms. Because increased glycolysis is an early feature of T-cell activation, we hypothesized that in vivo metabolic imaging of glycolysis would allow noninvasive detection of liver GVHD as activated CD4+ T cells traffic into the organ. Indeed, hyperpolarized 13C-pyruvate magnetic resonance imaging detected high rates of conversion of pyruvate to lactate in the liver ahead of animals becoming symptomatic, but not during subsequent overt chronic GVHD. Concomitantly, CD4+ T effector memory cells, the predominant pathogenic CD4+ T-cell subset, were confirmed to be highly glycolytic by transcriptomic, protein, metabolite, and ex vivo metabolic activity analyses. Preliminary data from single-cell sequencing of circulating T cells in patients undergoing AHSCT also suggested that increased glycolysis may be a feature of incipient acute GVHD. Metabolic imaging is being increasingly used in the clinic and may be useful in the post-AHSCT setting for noninvasive early detection of GVHD.

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31

White, James G. "Platelet interior imaging technologies." Blood 116, no. 26 (December 23, 2010): 6150–51. http://dx.doi.org/10.1182/blood-2010-09-305557.

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32

Ozpolat, Hasan Tahsin, Tim Chang, Junmei Chen, Xiaoping Wu, Colette Norby, Barbara A. Konkle, and Jose A. Lopez. "Evaluation of Cell Types and Morphologies in Sickle Cell Disease with an Imaging Flow Cytometer." Blood 126, no. 23 (December 3, 2015): 972. http://dx.doi.org/10.1182/blood.v126.23.972.972.

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Abstract Sickle cell disease (SCD) is a hemoglobinopathy characterized by vaso-occlusive episodes and hemolysis. Hemoglobin S is prone to polymerize at low oxygen tension, causing the red cell to become sickle shaped, more rigid and sticky. Evaluation of blood cell morphology, counts and activation are important components of the patient evaluation. This is usually accomplished by evaluation of the blood film, performing a complete blood count (CBC), and with the use of flow cytometry. A typical blood film from an SCD patient shows anisocytosis, poikilocytosis, polychromasia, nucleated erythrocytes, sickled cells, and irregular contracted cells. The methods of blood cell evaluation all have disadvantages. Preparation of the blood film is laborious and its evaluation is highly subjective and requires extensive experience. Some CBC counters (e.g., Siemens - ADVIA 2120) are able to detect dense cells (increased hemoglobin content-high MCHC cells) by their volume and hemoglobin concentration after the red blood cells (RBC) are swelled to spheres with a hypotonic solution. Dense cells resist becoming spheres and are detected by their low volume and high hemoglobin concentration. However, the number of dense cells might be underestimated because reversibly sickled cells are capable of undergoing the sphering and will not be detected. In addition, the hypotonic solution can lyse the cells. Finally, RBC counters cannot detect cells on the basis of specific cell markers, which can be used to define cell types and cell morphology and activation status (platelets). Conventional flow cytometry can detect cell markers, but yields little information on morphology and cannot detect dense cells. Here, we used the ImagestreamX Flow Cytometer (Amnis) to analyze SCD blood. In addition to providing information available with conventional cytometers, this instrument provides an image of each cell analyzed, thus allowing for detailed morphological assessment of a large population of cells. We analyzed 5 patients. All were outpatients not suffering from acute complications. Blood was collected by venipuncture into citrate anticoagulant, stained with antibodies or other reagents, and then fixed in 4% paraformaldehyde. We evaluated the blood for cell numbers and morphology, reticulocytes, dense cells, platelet-monocyte aggregates, phosphatidylserine exposure, and platelet activation status. The blood from all of the SCD patients displayed characteristics not found in control blood. We could clearly distinguish RBC morphologies corresponding to sickle cells, dense cell and reticulocytes. Reticulocytes, identified by CD71 positivity, often displayed a "puckered" morphology, as previously seen in electron micrographs. We calculated the percentage of RBCs that were sickled based on the shape ratio of > 2 (length along the long axis/maximum thickness along the short axis). The sickle cell percentage was 1.4±0.5% (normal 0%) out of total RBC population. We also evaluated dense cell morphology after separating the cells on a percoll density gradient. The cells appeared flattened and "deflated", clearly indicating their loss of intracellular fluid. We also analyzed platelet activation status based on staining for P-selectin, the activated form of integrin aIIbb3 (PAC-1 antibody), and phosphatidylserine exposure. Platelets staining positively for these markers also demonstrated morphological evidence of activation: shape change and filopodia extension. Platelet-monocyte aggregates were higher in all of the patients than in controls (0.036% vs 0%) and were easily distinguished from coincident events by morphology. The number of platelets associated with individual monocytes varied from 1 to 3. Other heterotypic cell aggregates were rare. In summary, imaging flow cytometry provides a powerful tool for the analysis of blood in SCD. The technique allows cell population analysis like conventional cytometry, while yielding detailed morphological information on many parameters of relevance in the disease. Further, the morphological assessment avoids many of the potential artifacts arising from blood film preparation and allows an unbiased assessment of the results. Disclosures Konkle: Baxalta: Consultancy, Research Funding; Biogen: Consultancy, Research Funding; CSL Behring: Consultancy, Other: IDMC chair; Pfizer: Other: IDMC member; Octapharma: Research Funding; Novo Nordisk: Consultancy.

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Lindner, Heidrun, Ernst Holler, Ines Klauke, Georg W. Bornkamm, and Günther Einer. "489Irradiated peripheral blood mononuclear cells induce endothelial cell apoptosis." Radiotherapy and Oncology 40 (January 1996): S125. http://dx.doi.org/10.1016/s0167-8140(96)80498-1.

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Park, Han-Sang, Hillel Price, Silvia Ceballos, Jen-Tsan Chi, and Adam Wax. "Single Cell Analysis of Stored Red Blood Cells Using Ultra-High Throughput Holographic Cytometry." Cells 10, no. 9 (September 17, 2021): 2455. http://dx.doi.org/10.3390/cells10092455.

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Holographic cytometry is introduced as an ultra-high throughput implementation of quantitative phase imaging of single cells flowing through parallel microfluidic channels. Here, the approach was applied for characterizing the morphology of individual red blood cells during storage under regular blood bank conditions. Samples from five blood donors were examined, over 100,000 cells examined for each, at three time points. The approach allows high-throughput phase imaging of a large number of cells, greatly extending our ability to study cellular phenotypes using individual cell images. Holographic cytology images can provide measurements of multiple physical traits of the cells, including optical volume and area, which are observed to consistently change over the storage time. In addition, the large volume of cell imaging data can serve as training data for machine-learning algorithms. For the study here, logistic regression was used to classify the cells according to the storage time points. The analysis showed that at least 5000 cells are needed to ensure accuracy of the classifiers. Overall, results showed the potential of holographic cytometry as a diagnostic tool.

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Guo, Xi, Yanwen Zhang, Jianbo Liu, Xiaohai Yang, Jin Huang, Li Li, Lan Wan, and Kemin Wang. "Red blood cell membrane-mediated fusion of hydrophobic quantum dots with living cell membranes for cell imaging." Journal of Materials Chemistry B 4, no. 23 (2016): 4191–97. http://dx.doi.org/10.1039/c6tb01067a.

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A biomimetic route to fusion of hydrophobic quantum dots (QDs) with living cells for membrane imaging was proposed. Red blood cell membrane lipids acted as both an efficient surfactant to phase-transfer QDs and a fusion reagent to facilitate fusion with cell membranes.

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Cohen, Jonathon B., Madhusmita Behera, Carrie A. Thompson, and Christopher R. Flowers. "Evaluating surveillance imaging for diffuse large B-cell lymphoma and Hodgkin lymphoma." Blood 129, no. 5 (February 2, 2017): 561–64. http://dx.doi.org/10.1182/blood-2016-08-685073.

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Abstract Up to 50% of patients with Hodgkin lymphoma and diffuse large B-cell lymphoma will relapse, requiring additional therapy. Although surveillance imaging is commonly performed in clinical practice, its ability to identify asymptomatic relapses and improve survival for patients is not well defined. We evaluated the surveillance imaging role in relapse detection and reviewed its impact on survival for relapsed patients, and found that current imaging approaches do not detect most relapses prior to clinical signs and symptoms or improve survival.

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Swinkels, Maurice, Johan A. Slotman, Frank W. G. Leebeek, Jan Voorberg, Ruben Bierings, and Gerard Jansen. "Super-Resolution Immunofluorescence Imaging of Platelet Granules." Blood 134, Supplement_1 (November 13, 2019): 3613. http://dx.doi.org/10.1182/blood-2019-124353.

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Introduction Platelets play a central in hemostasis by facilitating thrombus growth at sites of vascular injury. During activation, platelets secrete a highly diverse cocktail of proteins from their granules, thereby enhancing the hemostatic response. How platelets organize their granules in a resting state as well as during activation and secretion is still poorly understood. We have developed a platform for quantitative analysis of platelet granules employing super-resolution microscopy. Using structured illumination microscopy (SIM), individual alpha- and dense-granule can be separated in platelets. Together with morphological analysis and other quantitative imaging data that can be processed in automatic analysis workflows using ImageJ software, this method yields novel insights in granule organization and has implications for both platelet biology and translational studies on patients with platelet defects. Methods Platelet were isolated from citrated whole blood from consenting healthy donors and select patients. Samples were immediately fixed in 2% paraformaldehyde, washed, and loaded on poly-L-lysine coated high-resolution coverslips. We performed indirect immunostainings for various antigens, including alpha-tubulin, GP1b, Von Willebrand Factor (VWF) and platelet factor 4. Samples were imaged on an Elyra PS1 (Zeiss) with a SIM module using five phases and five rotations. After image acquisition, samples were reconstructed using Zen Software. Reconstructed images were analyzed by ImageJ plugins combined with in house written macro's which allowed for assessment of platelet morphology (volume, surface and shape), granule morphology (counts, volume, surface, shape) and granule metrics (smallest distance between granules and closest distance to membrane). We analyzed platelets from five healthy donors for general variation as well as from selected patients with platelet defects, which allowed us to validate our analytic workflow. Statistics was performed using GraphPad Prism 8. Results We have first assessed the variation in granule content in healthy donors. Segmentation of single resting platelets was done on cells stained for GP1b or alpha-tubulin, while alpha granule content of platelets was assessed by staining for VWF. These analyses revealed that the granule number (defined as unique VWF-positive spots of fluorescence using an optimized threshold) was normally distributed around the average of 16.78 per platelet. While there is considerable variation in alpha granule number (SD = 7.641), the sensitivity of our analysis is retained by analyzing hundreds of platelets per donor (SEM = 0.6923). We then tested whether our analysis could pick up subtle differences in platelet granules. For this purpose, we analyzed platelets from a type 2A Von Willebrand's Disease (VWD) patient with a C1190R mutation, leading to defective multimerization of VWF. Morphological analysis of SIM images revealed that the number of VWF-positive alpha granules was similar when compared to controls (16.7±0.7; p>0.05). Alpha granules in this patient were smaller (granule volume = 42.9±0.9 vs. controls 207.7±9.2; p<0.0001) and irregularly shaped (granule sphericity = 0.47±0.002 vs. controls 0.72±0.01; p<0.0001). As a control, we stained for platelet factor 4. There were no differences in granule number (18.3±0.4 vs. controls 19.5±0.5; p>0.05), nor granule volume (p=0.78). Altogether, this suggests that there is less VWF located in the alpha-granules of the VWD patient which we confirmed by Western blot and confocal microscopy. Our results imply that the packaging of VWF into alpha granules is impaired in VWD patients carrying the C1190R mutation. Discussion We have developed an automated image analysis workflow that allows for evaluation of platelets and their granules in patients with VWD. Our findings show the utility of unbiased, high throughput analyses of platelets and platelet granules for the diagnosis of hemostatic disorders. Disclosures Leebeek: Shire/Takeda: Research Funding; Novo Nordisk: Consultancy; UniQure: Consultancy; CSL Behring: Research Funding; Sobi: Other: Travel grant; Shire/Takeda: Consultancy. Jansen:Novartis: Other: Travel funding; 3SBio: Other: Speaker fee; Celgene: Other: Travel funding.

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Herman, Peter, Basavaraju G. Sanganahalli, and Fahmeed Hyder. "Multimodal Measurements of Blood Plasma and Red Blood Cell Volumes during Functional Brain Activation." Journal of Cerebral Blood Flow & Metabolism 29, no. 1 (September 3, 2008): 19–24. http://dx.doi.org/10.1038/jcbfm.2008.100.

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As an alternative to functional magnetic resonance imaging (fMRI) with blood oxygenation level dependent (BOLD) contrast, cerebral blood volume (CBV)-weighted fMRI with intravascular contrast agents in animal models have become popular. In this study, dynamic measurements of CBV were performed by magnetic resonance imaging (MRI) and laser-Doppler flowmetry (LDF) in α-chloralose anesthetized rats during forepaw stimulation. All recordings were localized to the contralateral primary somatosensory cortex as revealed by BOLD at 11.7 T. Ultra-small superparamagnetic iron oxide (15mg/kg)—a plasma-borne MRI contrast agent with a half-life of several hours in blood circulation—was used to quantify changes in magnetic field inhomogeneity in blood plasma. The LDF backscattered laser light (805 nm), which reflects the amount of red blood cells, was used to measure alterations in the non-plasma compartment. Dynamic and layer-specific comparisons of the two CBV signals during functional hyperemia revealed excellent correlations (> 0.86). These results suggest that CBV measurements from either compartment may be used to reflect dynamic changes in total CBV. Furthermore, by assuming steady-state mass balance and negligible counter flow, these results indicate that volume hematocrit is not appreciably affected during functional activation.

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Odaka, Masao, Hyonchol Kim, Yoshiyasu Nakamura, Akihiro Hattori, Kenji Matsuura, Moe Iwamura, Yohei Miyagi, and Kenji Yasuda. "Size Distribution Analysis with On-Chip Multi-Imaging Cell Sorter for Unlabeled Identification of Circulating Tumor Cells in Blood." Micromachines 10, no. 2 (February 25, 2019): 154. http://dx.doi.org/10.3390/mi10020154.

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We report a change of the imaging biomarker distribution of circulating tumor cell (CTC) clusters in blood over time using an on-chip multi-imaging flow cytometry system, which can obtain morphometric parameters of cells and those clusters, such as cell number, perimeter, total cross-sectional area, aspect ratio, number of nuclei, and size of nuclei, as “imaging biomarkers”. Both bright-field (BF) and fluorescent (FL) images were acquired at 200 frames per second and analyzed within the intervals for real-time cell sorting. A green fluorescent protein-transfected prostate cancer cell line (MAT-LyLu-GFP) was implanted into Copenhagen rats, and the blood samples of these rats were collected 2 to 11 days later and measured using the system. The results showed that cells having BF area of 90 μm2 or larger increased in number seven days after the cancer cell implantation, which was specifically detected as a shift of the cell size distribution for blood samples of implanted rats, in comparison with that for control blood. All cells with BF area of 150 μm2 or larger were arranged in cell clusters composed of at least two cells, as confirmed by FL nucleus number and area measurements, and they constituted more than 1% of all white blood cells. These results indicate that the mapping of cell size distribution is useful for identifying an increase of irregular cells such as cell clusters in blood, and show that CTC clusters become more abundant in blood over time after malignant tumor formation. The results also reveal that a blood sample of only 50 μL is sufficient to acquire a stable size distribution map of all blood cells to predict the presence of CTC clusters.

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Sharma, Akshay, Matthew Scoggins, Ping Zou, Amanda Kennedy, Richard Lovins, Sanjay Verma, Stacy High, et al. "Reduced Intensity Hematopoietic Cell Transplantation Improves Cerebral Hemodynamics in Children with Sickle Cell Disease." Blood 138, Supplement 1 (November 5, 2021): 125. http://dx.doi.org/10.1182/blood-2021-144993.

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Abstract Introduction Allogeneic hematopoietic cell transplantation (HCT) is increasingly being utilized in patients with sickle cell disease (SCD) to prevent progressive organ dysfunction, including for primary or secondary stroke prevention. However, outcomes after HCT have mostly been evaluated using broad measures such as overall survival and event free survival with limited direct physiologic evidence of improved cerebral hemodynamics. Cerebral blood flow (CBF; mL blood/100 g tissue/min) is increased in SCD to compensate for chronic anemia, reduced oxygen carrying capacity and to maintain adequate oxygen delivery to the brain tissue. In SCD patients, CBF is inversely associated with intelligence quotient (Strouse JJ, et al. Blood. 2006;108(1):379-381.) and working memory (Prussien KV, et al. Stroke. 2021;52(5):1830-1834.) and has been shown to improve with blood transfusions (Guilliams KP, et al. Blood. 2018;131(9):1012-1021.). Assessment of CBF may help predict long-term neurocognitive outcomes in patients with SCD and assess therapeutic responses to therapies. Additionally, the high metabolic demand of the brain potentially makes assessment of CBF a sensitive predictor of future multiorgan damage in patients with SCD and hence may be useful to help determine eligibility of patients for curative therapies. Methods We performed anatomical and hemodynamic magnetic resonance imaging (MRI) of the brain prior to, and at 6 months after HCT in children with SCD undergoing a reduced intensity conditioning based HCT on a clinical study (NCT04362293). All patients received pretransplant conditioning with hydroxyurea, azathioprine, alemtuzumab, thiotepa and low dose total body irradiation (200-400 cGy). All patients received an unmanipulated mobilized peripheral blood derived hematopoietic stem and progenitor cell graft. One patient who received the graft from a haploidentical (HAPLO) donor also received post-transplant cyclophosphamide. Graft versus host disease prophylaxis comprised of sirolimus. A 3-dimensional (3D) T1 sequence was used for gray/white matter segmentation. 3D-pulsed arterial spin labeling (ASL) perfusion imaging (3 mm isotropic) was acquired to measure resting CBF. Mean CBF values were calculated from gray matter. Mean ± standard deviation and median values for various variables are presented. Results Four consecutive patients had 2 serial MRIs performed (one before HCT and another 6 months after HCT). Median age at HCT was 14.5 years. Indications for HCT, donor type and whether patients were receiving chronic transfusion therapy (CTT) prior to HCT are indicated in the Table. No patient had prior evidence of overt stroke. All patients engrafted and had >99% donor myeloid chimerism at the time of the post-HCT imaging. Mean pre-HCT hemoglobin was 8.7 ± 0.8 g/dL (median 8.6 g/dL) and it increased to 12.5 ± 2.1 g/dL (median 12.4 g/dL) at 6 months after HCT. All patients exhibited elevated resting CBF values prior to HCT (123.8 ± 32.2 mL blood/100 g tissue/min, median 112.8 mL blood/100 g tissue/min), that decreased following HCT (71.1 ± 32.1 mL blood/100 g tissue/min, median 72.8 mL blood/100 g tissue/min) (Table and Figure). None of the patients had any clinical neurological complications or imaging evidence of new infarcts or anatomical abnormalities in the follow up period. Conclusion Our preliminary results indicate that it is feasible to quantify CBF in patients with SCD before and after HCT using MRI. We demonstrate that cerebral hemodynamics improve after HCT in children with SCD, indicated by a decrease in CBF to near normal values (normal CBF ~80 mL blood/100 g tissue/min). The more substantial changes (-50% and -78%) are seen in children who were not receiving CTT, suggesting that CTT might partially mitigate CBF elevation prior to HCT. Nevertheless, 2 patients receiving CTT prior to HCT also had a further modest decrease in CBF after HCT (-5% and -18%). We plan to follow these changes at serial time points annually after HCT and correlate them with changes in neurocognitive functioning and other functional MRI measures. Improved CBF is expected to reduce stroke risk and may also preserve or improve neurocognitive functioning, and prospective monitoring of these outcomes are underway. Data on all the study participants who have undergone an HCT and had 2 serial MRIs performed by the ASH annual meeting will be presented. Figure 1 Figure 1. Disclosures Sharma: Vertex Pharmaceuticals/CRISPR Therapeutics: Other: Salary support paid to institution; Medexus Inc: Consultancy; CRISPR Therapeutics: Other, Research Funding; Novartis: Other: Salary support paid to institution; Spotlight Therapeutics: Consultancy; Vindico Medical Education: Honoraria. Triplett: Miltenyi: Other: Travel, meeting registration. Hankins: Vindico Medical Education: Consultancy; UpToDate: Consultancy; Global Blood Therapeutics: Consultancy; Bluebird Bio: Consultancy.

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van Dam, Lisette F., Charlotte E. A. Dronkers, Gargi Gautam, Åsa Eckerbom, Waleed Ghanima, Jostein Gleditsch, Anders von Heijne, et al. "Magnetic resonance imaging for diagnosis of recurrent ipsilateral deep vein thrombosis." Blood 135, no. 16 (April 16, 2020): 1377–85. http://dx.doi.org/10.1182/blood.2019004114.

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Abstract The diagnosis of recurrent ipsilateral deep vein thrombosis (DVT) is challenging, because persistent intravascular abnormalities after previous DVT often hinder a diagnosis by compression ultrasonography. Magnetic resonance direct thrombus imaging (MRDTI), a technique without intravenous contrast and with a 10-minute acquisition time, has been shown to accurately distinguish acute recurrent DVT from chronic thrombotic remains. We have evaluated the safety of MRDTI as the sole test for excluding recurrent ipsilateral DVT. The Theia Study was a prospective, international, multicenter, diagnostic management study involving patients with clinically suspected acute recurrent ipsilateral DVT. Treatment of the patients was managed according to the result of the MRDTI, performed within 24 hours of study inclusion. The primary outcome was the 3-month incidence of venous thromboembolism (VTE) after a MRDTI negative for DVT. The secondary outcome was the interobserver agreement on the MRDTI readings. An independent committee adjudicated all end points. Three hundred five patients were included. The baseline prevalence of recurrent DVT was 38%; superficial thrombophlebitis was diagnosed in 4.6%. The primary outcome occurred in 2 of 119 (1.7%; 95% confidence interval [CI], 0.20-5.9) patients with MRDTI negative for DVT and thrombophlebitis, who were not treated with any anticoagulant during follow-up; neither of these recurrences was fatal. The incidence of recurrent VTE in all patients with MRDTI negative for DVT was 1.1% (95% CI, 0.13%-3.8%). The agreement between initial local and post hoc central reading of the MRDTI images was excellent (κ statistic, 0.91). The incidence of VTE recurrence after negative MRDTI was low, and MRDTI proved to be a feasible and reproducible diagnostic test. This trial was registered at www.clinicaltrials.gov as #NCT02262052.

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Scanlon, Vanessa M., Maria Kochugaeva, Juliana Xavier-Ferrucio, Yi-Chien Lu, Nayoung Kwon, Anisha Laumas, Matthew Cenci, et al. "Developing Single Cell Live Imaging Strategies to Determine MEP Fate and Predict Potential." Blood 134, Supplement_1 (November 13, 2019): 1190. http://dx.doi.org/10.1182/blood-2019-131204.

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The molecular mechanisms underlying lineage commitment of stem and progenitor cells have implications for deriving specific cell types in vitro for regenerative medicine purposes and elucidating the aberrant pathways responsible for pathological conditions. We investigated Megakaryocytic-Erythroid Progenitors (MEP) commitment to the megakaryocytic (Mk) and erythroid (E) lineages as a model of cell fate decisions. Colony forming unit (CFU) assays are used to test the functional output, or lineage potential, of progenitor cell populations. As single progenitor cells proliferate, their progeny remain near each other to form a colony. This potential is deduced from the mature cell types comprising the colony. However, this assay has several limitations: 1) there is an assumption that each colony arose from a single progenitor cell based on the low probability that individual progenitors would form overlapping colonies when plated sparsely; 2) there is a surfeit of kinetic data about the colony forming cells that is not collected with a single end-point; and 3) the outcome may be misleading if there is selective loss of specific cell types of early committed progeny prior to the endpoint. To overcome these limitations, we developed a time-lapse microscopy and lineage tracing approach to visualize single sorted MEP, as well as committed Megakaryocytic progenitors (MkP) and Erythroid progenitors (ErP), as they proliferate, specify and progress down either the Mk or E lineages. We plated primary adult human MEP (Sanada and Xavier-Ferrucio, et al. Blood, 2016) at low density in semisolid media supplemented with cytokines in a MatTek plate under a coverslip to minimize focal range and support colony formation for up to 14 days. The plated cells were imaged with an Olympus VivaView. Fluorescently conjugated anti-CD41 (a marker of Mk) and anti-CD235a (a marker of E) were added to the culture dishes at the end to confirm the lineage of each cell in the colonies. The average time from plating to the first division was 38 h, and not significantly different between MEP, ErP, and MkP. We optimized the image acquisition settings to permit accurate tracking of the plated cells and their progeny for up to 11 days. Average colony forming efficiency was 75%, which was equal to unimaged CFU assays, indicating minimal phototoxicity. Acquired images were stacked into time-lapse videos and automatically tracked with the Baxter Algorithm (Magnusson et al. IEEE Trans Med Imaging, 2015). After manual segmentation and track correction, lineage trees were generated for each colony (Figure 1). We exported quantifiable features of the cells including the time between cell divisions and velocity of each cell from the algorithm. We defined cells that give rise to only CD41-labeled cells as committed MkP, cells that give rise to only CD235-labeled cells as ErP, and cells that give rise to at least one progeny of both as MEP. However, these progenitor definitions are based on an inaccurate assumption that fate reflects potential, since a bipotent cell may stochastically give rise to progeny that are committed to a single lineage. To convert fate to potential, we first need to identify the precise period of commitment to more accurately define MEP, MkP, and ErP in our culture system. To achieve this, we are developing a mathematical model that can predict the potential of cells based on quantifiable features that are measured form the time-lapse images and are significantly different between MEP, MkP, and ErP. Utilizing statistical tools, we have identified that the duration of the cell cycle and the velocity of single progenitor cells along with their lineage history, permit predictions of progenitor potential and provide several novel insights into the process of lineage commitment. We found that ErP have a significantly shortened cell cycle duration and slower velocity compared to MkP, while MEP have the longest cell cycle and an intermediate velocity until undergoing their fate decisions (Figures 2 and 3). With this time-lapse imaging approach, we hope to better investigate molecular mechanisms that direct fate decisions of multipotent progenitor cells. Disclosures No relevant conflicts of interest to declare.

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YOO, JAMES H. K., JOSEPH A. ORZEL, JAMES W. BAGNALL, and FREDERICK L. WEILAND. "Technetium-99m Red Blood Cell Blood-pool Imaging in Functional Asplenia Due to Leukemic Infiltration." Clinical Nuclear Medicine 11, no. 7 (July 1986): 493–94. http://dx.doi.org/10.1097/00003072-198607000-00007.

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Kelb�k, Henning, Kirsten B�low, Jan Aldershvile, Jens M�gelvang, and Steen Levin Nielsen. "A new 99mTc-red blood cell labeling procedure for cardiac blood pool imaging: clinical results." European Journal of Nuclear Medicine 15, no. 7 (July 1989): 333–35. http://dx.doi.org/10.1007/bf00449219.

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Simonetta, Federico, Israt S. Alam, Juliane K. Lohmeyer, Bita Sahaf, Zinaida Good, Weiyu Chen, Zunyu Xiao, et al. "Molecular Imaging of Chimeric Antigen Receptor T Cells By ICOS-Immunopet." Blood 136, Supplement 1 (November 5, 2020): 5–6. http://dx.doi.org/10.1182/blood-2020-136331.

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Introduction: Immunomonitoring of chimeric antigen receptor (CAR) T cells relies primarily on their quantification in the peripheral blood, which inadequately quantifies their biodistribution and activation status in the tissues. Non-invasive molecular imaging of CAR T cell therapy by positron emission tomography (PET) is a promising approach providing spatial, temporal and functional information. Reported strategies for PET-based monitoring of CAR T cells rely on additional manipulation of the cell product such as the incorporation of reporter transgenes or ex vivo biolabeling, which significantly limits the wider application of CAR T cell molecular imaging. In the present study, we assessed the ability of antibody-based PET (immunoPET) to non-invasively visualize CAR T cells in vivo. Methods: For analysis of human CAR T cell activation, we analyzed publicly available RNA sequencing data (GSE136891) obtained at serial time points during in vitro culture of CD19.CD28z CAR T cells. We analyzed by mass cytometry (CyTOF) the ex vivo ICOS expression on human CD19-28z CAR T cells obtained from 31 patients receiving axicabtagene ciloleucel (Axi-cel) for relapsed/refractory diffuse large B-cell lymphoma (DLBCL). For in vivo murine experiments, CD19-expressing B-cell lymphoma A20 cells (2.5×10e5 cells) were injected by tail vein intravenously (i.v.) into sub-lethally (4.4 Gy) irradiated Thy1.2+ BALB/c mice. Seven days later, murine CD19.CD28z Luc+ Thy1.1+ CAR T cells (1×10e6) were i.v. injected. ICOS expression was analyzed by flow cytometry on CAR T cells recovered from spleen and bone marrow 5 days after injection. For imaging studies, anti-ICOS monoclonal antibody (mAb) specific for murine ICOS (clone:7E.17G9, BioXcell) was modified with the bifunctional chelator deferoxamine (DFO/p-SCN-Bn-Deferoxamine). The DFO-ICOS mAb conjugate was radiolabeled with 37 MBq (~1 mCi) of 89Zr-oxalate (final specific activity 6 µCi/µg/ml and radiochemical purity of 99%). 89Zr-DFO-ICOSmAb (45 μCi ± 3.6, 7.5 μg± 0.6) was injected i.v. 5 days post-CAR T cell administration and PET-CT imaging performed 48 hours later. Following PET-CT, mice were euthanized and radioactivity measured in dissected weighed tissues using a gamma-counter. Results: Analysis of RNA-sequencing data from human CAR T cells identified ICOS as an activation marker whose transcription was up-regulated and sustained during in vitro culture. ICOS was preferentially expressed on CAR+ T cells recovered at day 7 from axi-cel treated patients compared with CAR- cells (p<0.001; Figure 1A). Phenotypic analysis in a murine model of B cell lymphoma infiltrating the spleen and the bone marrow confirmed preferential ICOS expression on murine CAR T cells compared to resident cells in both spleen (p=0.003) and bone marrow (p=0.008). Figure 1B shows representative volume-rendered technique (VRT) PET/CT images of 89Zr-DFO-ICOS mAb-injected tumor-bearing mice either untreated (left panels) or that received mCD19.28z CAR T cells (right panels). 89Zr-DFO-ICOS mAb similarly accumulated in highly vascularized organs (heart, liver and spleen) of both untreated and CAR T cell treated mice, consistent with the biodistribution and clearance of intact antibodies. We detected pronounced 89Zr-DFO-ICOS mAb-PET signals in the bones of CAR T cell treated mice, particularly prominent in the lumbar spine, iliac bones, femur, tibia and humeral heads (Figure 1B). Region of interest analysis confirmed markedly increased radiotracer uptake in bones rich in bone marrow from CAR T treated mice compared with those of untreated mice (lumbar spine vertebrae p<0.001; iliac bones p=0.001; femur p=0.002; tibia p=0.002). Moreover we observed a slight, but statistically significant increase in radiotracer accumulation in the heart of CAR T cell-treated mice (p=0.004) while no significant differences were detected in spleen and liver. As expected, there was no significant signal difference in the muscle, considered background. Biodistribution analysis using gamma counting of tissues confirmed the PET results. Conclusions: We describe for the first time an immunoPET approach to monitor the in vivo dynamics of CAR T cell migration, expansion, and persistence that does not require the addition of reporter genes or ex vivo labeling, being therefore applicable to the clinical setting for the study of any commercially available and investigational CAR T cell products. Disclosures Miklos: Novartis: Consultancy, Other: Travel support, Research Funding; Allogene Therapeutics Inc.: Research Funding; Pharmacyclics: Consultancy, Other: Travel support, Patents & Royalties, Research Funding; Juno-Celgene-Bristol-Myers Squibb: Consultancy, Other: Travel support, Research Funding; Janssen: Consultancy, Other: Travel support; Miltenyi Biotec: Research Funding; Kite-Gilead: Consultancy, Membership on an entity's Board of Directors or advisory committees, Other: Travel support, Research Funding; Adaptive Biotech: Consultancy, Other: Travel support, Research Funding. Mackall:BMS: Consultancy; Allogene: Current equity holder in publicly-traded company; Lyell Immunopharma: Consultancy, Current equity holder in private company; NeoImmune Tech: Consultancy; Nektar Therapeutics: Consultancy; Apricity Health: Consultancy, Current equity holder in private company. Gambhir:CellSight Inc: Current equity holder in private company. Negrin:Amgen: Consultancy; BioEclipse Therapeutics: Current equity holder in private company; Magenta Therapeutics: Consultancy, Current equity holder in publicly-traded company; Biosource: Current equity holder in private company; KUUR Therapeutics: Consultancy; UpToDate: Honoraria.

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46

George, Thaddeus, Brian Hall, Sherree Friend, Scott Mordecai, Richard DeMarco, and Raymond Kong. "Analysis of whole blood immune cell function using imaging flow cytometry. (65.8)." Journal of Immunology 186, no. 1_Supplement (April 1, 2011): 65.8. http://dx.doi.org/10.4049/jimmunol.186.supp.65.8.

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Abstract Analysis of immune function directly in human whole blood samples has several advantages over analysis in processed samples. First, leucocytes are activated in conditions closely matching their natural environment in the bloodstream, complete with all cofactors and cell types necessary for normal function. Second, purification processes are time consuming, technically challenging, and can themselves be a source of stress (temperature, physical, pH, etc) that modulates immune cell function. Furthermore, whole blood is the most obtainable source of clinically-relevant material from patients and healthy controls. Whole blood imaging assays are challenging due to the need for simultaneous multispectral immunophenotyping and in many cases due to the rarity of the cell type of interest. However, recent advancements in high speed flow imaging uniquely enable the study of leukocyte function directly in whole blood samples in three image-based applications that we present here. These include measurement of: 1) chemokine-induced receptor internalization in whole blood T cells; 2) chemokine-induced shape change in whole blood monocytes; and 3) dose and time kinetic activation of NFkB translocation in monocytes and plasmacytoid dendritic cells, the latter of which represent less than 0.1% of whole blood leukocytes. These results uniquely demonstrate methods for statistically robust image-based measurement of rare immune cell function directly in whole blood samples.

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47

Chen, Yong, Lingyun Shao, Zahida Ali, Jiye Cai та Zheng W. Chen. "NSOM/QD-based nanoscale immunofluorescence imaging of antigen-specific T-cell receptor responses during an in vivo clonal Vγ2Vδ2 T-cell expansion". Blood 111, № 8 (15 квітня 2008): 4220–32. http://dx.doi.org/10.1182/blood-2007-07-101691.

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Abstract Nanoscale imaging of an in vivo antigen-specific T-cell immune response has not been reported. Here, the combined near-field scanning optical microscopy– and fluorescent quantum dot–based nanotechnology was used to perform immunofluorescence imaging of antigen-specific T-cell receptor (TCR) response in an in vivo model of clonal T-cell expansion. The near-field scanning optical microscopy/quantum dot system provided a best-optical-resolution (<50 nm) nano-scale imaging of Vγ2Vδ2 TCR on the membrane of nonstimulated Vγ2Vδ2 T cells. Before Ag-induced clonal expansion, these nonstimulating Vγ2Vδ2 TCRs appeared to be distributed differently from their αβ TCR counterparts on the cell surface. Surprisingly, Vγ2Vδ2 TCR nanoclusters not only were formed but also sustained on the membrane during an in vivo clonal expansion of Vγ2Vδ2 T cells after phosphoantigen treatment or phosphoantigen plus mycobacterial infection. The TCR nanoclusters could array to form nanodomains or microdomains on the membrane of clonally expanded Vγ2Vδ2 T cells. Interestingly, expanded Vγ2Vδ2 T cells bearing TCR nanoclusters or nanodomains were able to rerecognize phosphoantigen and to exert better effector function. These studies provided nanoscale insight into the in vivo T-cell immune response.

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48

Gao, Chuang, Pravin Bhattarai, Yiming Zhou, Nisi Zhang, Sadaf Hameed, Xiuli Yue, and Bo Zhao. "Harnessing platelets as functional vectors for contrast enhanced ultrasound imaging and fluorescence imaging." RSC Advances 9, no. 72 (2019): 41993–99. http://dx.doi.org/10.1039/c9ra05118j.

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49

Wang, Weijia, Yang Zhang, Philip Dettinger, Andreas Reimann, Tobias Kull, Dirk Loeffler, Markus G. Manz, Claudia Lengerke, and Timm Schroeder. "Cytokine combinations for human blood stem cell expansion induce cell-type– and cytokine-specific signaling dynamics." Blood 138, no. 10 (May 14, 2021): 847–57. http://dx.doi.org/10.1182/blood.2020008386.

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Abstract How hematopoietic stem cells (HSCs) integrate signals from their environment to make fate decisions remains incompletely understood. Current knowledge is based on either averages of heterogeneous populations or snapshot analyses, both missing important information about the dynamics of intracellular signaling activity. By combining fluorescent biosensors with time-lapse imaging and microfluidics, we measured the activity of the extracellular-signal–regulated kinase (ERK) pathway over time (ie, dynamics) in live single human umbilical cord blood HSCs and multipotent progenitor cells (MPPs). In single cells, ERK signaling dynamics were highly heterogeneous and depended on the cytokines, their combinations, and cell types. ERK signaling was activated by stem cell factor (SCF) and FMS-like tyrosine kinase 3 ligand in HSCs but SCF, interleukin 3, and granulocyte colony-stimulating factor in MPPs. Different cytokines and their combinations led to distinct ERK signaling dynamics frequencies, and ERK dynamics in HSCs were more transient than those in MPPs. A combination of 5 cytokines recently shown to maintain HSCs in long-term culture, had a more-than-additive effect in eliciting sustained ERK dynamics in HSCs. ERK signaling dynamics also predicted future cell fates. For example, CD45RA expression increased more in HSC daughters with intermediate than with transient or sustained ERK signaling. We demonstrate heterogeneous cytokine- and cell-type–specific ERK signaling dynamics, illustrating their relevance in regulating hematopoietic stem and progenitor (HSPC) cell fates.

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50

Takahashi, Miwako, Sodai Takyu, Aya Sugyo, Hitomi Sudo, Hideaki Tashima, Hidekatsu Wakizaka, Go Akamatsu, et al. "Visualization of Multiple Myeloma Mouse Model with 89zr-Labeled Anti-CD38 Antibody By a New Imaging System; Whole-Gamma Imaging." Blood 140, Supplement 1 (November 15, 2022): 12476–77. http://dx.doi.org/10.1182/blood-2022-165050.

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