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New York Blood Center

2 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Protocol for Recombinant RBD-based SARS Vaccines: Protein Preparation, Animal Vaccination and Neutralization Detection

Lanying Du¹, Xiujuan Zhang¹, Jixiang Liu¹, Shibo Jiang¹

¹Lindsley F. Kimball Research Institute, New York Blood Center

This protocol describes a general procedure for studying recombinant receptor-binding domain (RBD)-based subunit vaccines against SARS. It includes methods for transfection and expression of RBD protein in 293T cells, immunization of mice with RBD and detection of neutralization activity of mouse sera using an established SARS pseudovirus neutralization assay.

Immunology and Infection

Measuring Deformability and Red Cell Heterogeneity in Blood by Ektacytometry

Nermi L. Parrow^*1, Pierre-Christian Violet^*2, Hongbin Tu², James Nichols³, Corinne A. Pittman⁴, Courtney Fitzhugh⁴, Robert E. Fleming^1,5, Narla Mohandas⁶, John F. Tisdale³, Mark Levine²

¹Department of Pediatrics, Saint Louis University School of Medicine, ²Molecular and Clinical Nutrition Section, Digestive Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, ³Molecular and Clinical Hematology Branch, National Institute of Diabetes and Digestive and Kidney Diseases, ⁴Sickle Cell Branch, National Heart, Lung and Blood Institute, National Institutes of Health, ⁵Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, ⁶Red Cell Physiology Laboratory, New York Blood Center

Here we present techniques to measure red cell deformability and cellular heterogeneity by ektacytometry. These techniques are applicable to general investigations of red cell deformability and specific investigations of blood diseases characterized by the presence of both rigid and deformable red cells in circulation, such as sickle cell anemia.