This readme.txt file was generated on 20230713 by Pampusch (edited by data curator 20230828) Recommended citation for the data: Davey, Brianna, C; Pampusch, Mary, S; Cartwright, Emily, K; Abdelaal, Hadia, M; Rakasz, Eva, G; Rendahl, Aaron; Berger, Edward, A; Skinner, Pamela, J. (2022). Anti-CAR antibody response in SIV infected rhesus macaques. Retrieved from the Data Repository for the University of Minnesota, https://doi.org/10.13020/d3e7-wt82. ------------------- GENERAL INFORMATION ------------------- 1. Title: Anti-CAR antibody response in SIV infected rhesus macaques 2. Authors: Brianna C Davey, Mary S Pampusch, Emily K Cartwright, Hadia M Abdelaal, EVA G RAKASZ, Aaron Rendahl, Edward A Berger, Pamela J Skinner Principal Investigator Contact Information Name:Pamela J Skinner Institution: University of Minnesota Address: 2-107 MRF, 689 23rd Ave SE, Minneapolis, MN 55455 Email: skinn002@umn.edu ORCID: 0000-0003-3388-1687 Associate or Co-investigator Contact Information Name: Brianna C Davey Institution: University of Minnesota Address: 689 23rd Ave SE, Minneapolis, MN 55455 Email: davey042@umn.edu ORCID: Associate or Co-investigator Contact Information Name: Mary S Pampusch Institution: University of Minnesota Address: 2-139 MRF, 689 23rd Ave SE, Minneapolis, MN 55455 Email: pampu002@umn.edu ORCID: 0000-0002-6973-9474 3. Date published or finalized for release: 20221122 4. Date of data collection (single date, range, approximate date) 12/01/2020 - 06/30/2022 5. Geographic location of data collection (where was data collected?): University of Minnesota 6. Information about funding sources that supported the collection of the data: This research was supported by: 5R01AI096966-06S1 (PS, and EB), 1UM1AI26617 (PS, and EB), P51OD011106/ P51RR000167 (ER), R01AI143380 (PS and EB), 1UM14126617 (PS and EC), training grant 5- T32 AI 55433 (EKC) as well as funds provided by the NIAID Division of Intramural Research (https:// www.niaid.nih.gov) and the NIH Intramural AIDS Targeted Antiviral Program (https://irp.nih.gov) (EB). 7. Overview of the data (abstract): T cells expressing a simian immunodeficiency (SIV)-specific chimeric antigen receptor (CAR) and the follicular homing molecule, CXCR5, were infused into antiretroviral therapy (ART) suppressed, SIV-infected rhesus macaques to assess their ability to localize to the lymphoid follicle and control the virus upon ART interruption. The cells did not persist in the animals beyond 28 days. Development of anti-CAR antibodies could be responsible for the lack of persistence. Potential antigenic sites on the anti-SIV CAR used in these studies included domains 1 and 2 of CD4, the carbohydrate recognition domain (CRD) of mannose-binding lectin (MBL), and an extracellular domain of the costimulatory molecule, CD28, along with short linker sequences. Using a flow cytometry based assay and target cells expressing the CAR/CXCR5 construct, we examined the serum of the CAR T-cell treated animals to determine that the animals had developed an anti-CAR antibody response after infusion. Binding sites for the anti-CAR antibodies were identified by using alternative CARs transduced into target cells and by preincubation of the target cells with a CD4 blocking antibody. All of the treated animals developed antibodies in their serum that bound to CAR-T cells and the majority were capable of inducing an ADCC response. The CD4 antibody-blocking assay suggests that the dominant immunogenic components of this CAR are the CD4 domains with a possible additional site of the CD28 domain with its linker. -------------------------- SHARING/ACCESS INFORMATION -------------------------- 1. Licenses/restrictions placed on the data: CC0 1.0 Universal https://creativecommons.org/publicdomain/zero/1.0/ 2. Links to publications that cite or use the data: Davey, B. C., Pampusch, M. S., Cartwright, E. K., Abdelaal, H. M., Rakasz, E. G., Rendahl, A., ... & Skinner, P. J. (2022). Development of an anti-CAR antibody response in SIV-infected rhesus macaques treated with CD4-MBL CAR/CXCR5 T cells. Frontiers in immunology, 13, 1032537. 3. Was data derived from another source? no 4. Terms of Use: Data Repository for the U of Minnesota (DRUM) By using these files, users agree to the Terms of Use. https://conservancy.umn.edu/pages/drum/policies/#terms-of-use --------------------- DATA & FILE OVERVIEW --------------------- 1. File List A. Davey_statistical analysis.xlsx Short description: Table with p values and confidence intervals from correlations of antibody level with other factors as presented in the manuscript B. Davey_Anti-CAR ab levels (figure 1, 2 and supplementary figure 1).xlsx Short description: File showing levels of antibody detected using different target cells and the serum of the different animals in the study C. Davey_CD4 Blocking data (figure 3).xlsx Short description: Antibody binding data in target cells blocked with anti-CD4 antibody D. Davey_ADCC data (figure 4).xlsx Short description: Data from antibody dependent cytotoxicity assays using serum from the treated animals E. Davey_CARCXCR5 studies (supplemental figure 3).xlsx Short description: Information about the cells infused into the treatment animals leading to antibody generation F. Davey_AllCorrelationData (figure 5 and supplementary figure 4).xlsx Short description: Data used in running correlations between antibody levels and other factors such as persistence, area under the curve and MBL MFI. 2. Relationship between files: Files contain the raw data used to generate the figures in the manuscript as well as the statistical values generated. All relate to the levels of anti-CAR antibody generated in animals treated with CAR T cells and the functionality of those antibodies Note: Curator created a zipped file of csv copies of all files for preservation and interoperability purposes. These are duplicates of individual file list. -------------------------- METHODOLOGICAL INFORMATION -------------------------- 1. Description of methods used for collection/generation of data: Most of the data was generated by incubating serum from CAR-T treated animals with CAR-T cells and measuring binding by flow cytometry. We also present data from an antibody-dependent cellular cytotoxicity assay to demonstrate functionality. This assay is also flow cytometry based. 2. Methods for processing the data: The data was generated by comparing binding of serum samples from treated and untreated animals to the CAR T cells. Background was subtracted from the samples and then comparisons were made between animals or between different types of target cells. 3. Instrument- or software-specific information needed to interpret the data: Binding of the antibodies was measured on a Beckman Cytoflex flow cytometer with data analyzed by FlowJo v10. 4. Standards and calibration information, if appropriate: NA 5. Environmental/experimental conditions: NA 6. Describe any quality-assurance procedures performed on the data: NA 7. People involved with sample collection, processing, analysis and/or submission: Sample collection was carried out by Brianna Davey. Data for correlations was generated by Brianna Davey, Mary Pampusch and Hadia Abdellal Analysis was conducted by Brianna Davey, Emily Cartwright, Mary Pampusch and Aaron Rendahl Ethics statement: All rhesus macaques used in this study were cared for by the staff at the Wisconsin National Primate Research Center (WNPRC) in accordance with the principles described in the National Research Council's Guide for the Care and Use of Laboratory Animals and with the recommendations of the Weatherall report (https://royalsociety.org/topics-policy/publications/2006/weatherall-report/). This study was approved by the University of Wisconsin-Madison Graduate School Institutional Animal Care and Use Committee (Animal Care and Use Protocol Number G005529).