PMID- 38411617 OWN - NLM STAT- MEDLINE DCOM- 20240228 LR - 20240529 IS - 1540-9538 (Electronic) IS - 0022-1007 (Print) IS - 0022-1007 (Linking) VI - 221 IP - 4 DP - 2024 Apr 1 TI - Framework for in vivo T cell screens. LID - 10.1084/jem.20230699 [doi] LID - e20230699 AB - In vivo T cell screens are a powerful tool for elucidating complex mechanisms of immunity, yet there is a lack of consensus on the screen design parameters required for robust in vivo screens: gene library size, cell transfer quantity, and number of mice. Here, we describe the Framework for In vivo T cell Screens (FITS) to provide experimental and analytical guidelines to determine optimal parameters for diverse in vivo contexts. As a proof-of-concept, we used FITS to optimize the parameters for a CD8+ T cell screen in the B16-OVA tumor model. We also included unique molecular identifiers (UMIs) in our screens to (1) improve statistical power and (2) track T cell clonal dynamics for distinct gene knockouts (KOs) across multiple tissues. These findings provide an experimental and analytical framework for performing in vivo screens in immune cells and illustrate a case study for in vivo T cell screens with UMIs. CI - © 2024 Milling et al. FAU - Milling, Lauren E AU - Milling LE AUID- ORCID: 0009-0002-4604-9435 AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 FAU - Markson, Samuel C AU - Markson SC AUID- ORCID: 0000-0001-9053-9867 AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Broad Institute of Harvard and Massachusetts Institute of Technology , Cambridge, MA, USA. ROR: https://ror.org/05a0ya142 FAU - Tjokrosurjo, Qin AU - Tjokrosurjo Q AUID- ORCID: 0000-0003-0965-9461 AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 FAU - Derosia, Nicole M AU - Derosia NM AUID- ORCID: 0009-0002-9429-8946 AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 FAU - Streeter, Ivy S L AU - Streeter ISL AUID- ORCID: 0000-0003-0729-1042 AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 FAU - Hickok, Grant H AU - Hickok GH AUID- ORCID: 0009-0004-8235-4773 AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 FAU - Lemmen, Ashlyn M AU - Lemmen AM AUID- ORCID: 0009-0002-9296-7210 AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 FAU - Nguyen, Thao H AU - Nguyen TH AUID- ORCID: 0000-0002-2767-0528 AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 FAU - Prathima, Priyamvada AU - Prathima P AUID- ORCID: 0000-0002-9907-4696 AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 FAU - Fithian, William AU - Fithian W AUID- ORCID: 0000-0003-2331-3029 AD - Department of Statistics, University of California, Berkeley, Berkeley, CA, USA. ROR: https://ror.org/01an7q238 FAU - Schwartz, Marc A AU - Schwartz MA AUID- ORCID: 0000-0003-3869-0150 AD - Broad Institute of Harvard and Massachusetts Institute of Technology , Cambridge, MA, USA. ROR: https://ror.org/05a0ya142 AD - Department of Medicine, Massachusetts General Hospital Cancer Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 FAU - Hacohen, Nir AU - Hacohen N AUID- ORCID: 0000-0002-2349-2656 AD - Broad Institute of Harvard and Massachusetts Institute of Technology , Cambridge, MA, USA. ROR: https://ror.org/05a0ya142 AD - Department of Medicine, Massachusetts General Hospital Cancer Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 FAU - Doench, John G AU - Doench JG AUID- ORCID: 0000-0002-3707-9889 AD - Broad Institute of Harvard and Massachusetts Institute of Technology , Cambridge, MA, USA. ROR: https://ror.org/05a0ya142 FAU - LaFleur, Martin W AU - LaFleur MW AUID- ORCID: 0000-0002-5017-774X AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Broad Institute of Harvard and Massachusetts Institute of Technology , Cambridge, MA, USA. ROR: https://ror.org/05a0ya142 FAU - Sharpe, Arlene H AU - Sharpe AH AUID- ORCID: 0000-0002-9736-2109 AD - Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Gene Lay Institute of Immunology and Inflammation, Brigham and Women's Hospital, Massachusetts General Hospital and Harvard Medical School , Boston, MA, USA. ROR: https://ror.org/03wevmz92 AD - Broad Institute of Harvard and Massachusetts Institute of Technology , Cambridge, MA, USA. ROR: https://ror.org/05a0ya142 LA - eng GR - P01 AI108545/AI/NIAID NIH HHS/United States GR - U19 AI133524/AI/NIAID NIH HHS/United States PT - Journal Article DEP - 20240227 PL - United States TA - J Exp Med JT - The Journal of experimental medicine JID - 2985109R SB - IM MH - Animals MH - Mice MH - *CD8-Positive T-Lymphocytes MH - Gene Knockout Techniques PMC - PMC10899089 COIS- Disclosures: L.E. Milling and M.W. LaFleur reported grants from Merck Sharp & Dohme LLC during the conduct of the study. N.M. Derosia and P. Prathima reported grants from Merck during the conduct of the study and grants from Merck outside the submitted work. I.S.L. Streeter and A.M. Lemmen reported grants from Merck during the conduct of the study. G.H. Hickok reported grants from Merck & Co. during the conduct of the study. N. Hacohen reported personal fees from Danger Bio/Related Science, Immune Repertoire Medicines, and CytoReason, grants from Bristol Myers Squibb, and grants from Calico Life Sciences outside the submitted work. J.G. Doench reported personal fees from BioNTech, Tango Therapeutics, Microsoft Research, and Pfizer outside the submitted work; in addition, J.G. Doench consults for Microsoft Research, Abata Therapeutics, Servier, Maze Therapeutics, BioNTech, Sangamo, and Pfizer; consults for and has equity in Tango Therapeutics; serves as a paid scientific advisor to the Laboratory for Genomics Research, funded in part by GlaxoSmithKline; and receives funding support from the Functional Genomics Consortium: Abbvie, Bristol Myers Squibb, Janssen, Vir Biotechnology, and Merck Sharp & Dohme LLC, a subsidiary of Merck & Co., Inc., Rahway, NJ, USA. J.G. Doench’s interests were reviewed and managed by the Broad Institute in accordance with its conflict-of-interest policies. A.H. Sharpe reported grants from NIH U19AI133524 and grants from NIH P01 AI108545 during the conduct of the study; grants from Vertex, Moderna, Merck Sharp & Dohme, AbbVie, Quark/IOME, Roche, Ipsen, Novartis, Erasca, Taiwan Bio, and Calico; personal fees from Surface Oncology, Sqz Biotechnologies, Elpiscience, Selecta, Bicara, Fibrogen, Alixia, GlaxoSmithKline, Janssen, Amgen, and Bioentre; and “other” from IOME, Monopteros, and Corner Therapeutics outside the submitted work; in addition, A.H. Sharpe had patent numbers 7,432,059 and 7,722,868 with royalties paid “Roche, Merck, Bristol-Myers-Squibb, EMD-Serono, Boehringer-Ingelheim, AstraZeneca, Leica, Mayo Clinic, Dako and Novartis,” patent numbers 8,652,465 and 9,457,080 licensed “Roche,” patent numbers 9,683,048, 9,815,898, 9,845,356, 10,202,454, and 10,457,733 licensed “Novartis,” patent numbers 9,580,684, 9,988,452, 10,370,446, 10,457,733, 10,752,687, 10,851,165, 10,934,353, and 15,314,251 issued; and is on scientific advisory boards for the Massachusetts General Cancer Center, Program in Cellular and Molecular Medicine at Boston Children’s Hospital, the Human Oncology and Pathogenesis Program at Memorial Sloan Kettering Cancer Center, The Gladstone Institute, and the Bloomberg-Kimmel Institute for Cancer Immunotherapy is an academic editor for the Journal of Experimental Medicine. A.H. Sharpe is on advisory boards for Elpiscience, Bicara, Monopteros, Fibrogen, Corner Therapeutics, Bioentre, IOME, Alixia, GlaxoSmithKline, Janssen, and Amgen. No other disclosures were reported. EDAT- 2024/02/27 12:43 MHDA- 2024/02/28 06:45 PMCR- 2024/08/27 CRDT- 2024/02/27 10:54 PHST- 2023/04/24 00:00 [received] PHST- 2023/12/14 00:00 [revised] PHST- 2024/01/19 00:00 [accepted] PHST- 2024/08/27 00:00 [pmc-release] PHST- 2024/02/28 06:45 [medline] PHST- 2024/02/27 12:43 [pubmed] PHST- 2024/02/27 10:54 [entrez] AID - 276584 [pii] AID - jem.20230699 [pii] AID - 10.1084/jem.20230699 [doi] PST - ppublish SO - J Exp Med. 2024 Apr 1;221(4):e20230699. doi: 10.1084/jem.20230699. Epub 2024 Feb 27. PMID- 38354702 OWN - NLM STAT- MEDLINE DCOM- 20240216 LR - 20240225 IS - 1097-4180 (Electronic) IS - 1074-7613 (Print) IS - 1074-7613 (Linking) VI - 57 IP - 2 DP - 2024 Feb 13 TI - The B7:CD28 family and friends: Unraveling coinhibitory interactions. PG - 223-244 LID - S1074-7613(24)00038-4 [pii] LID - 10.1016/j.immuni.2024.01.013 [doi] AB - Immune responses must be tightly regulated to ensure both optimal protective immunity and tolerance. Costimulatory pathways within the B7:CD28 family provide essential signals for optimal T cell activation and clonal expansion. They provide crucial inhibitory signals that maintain immune homeostasis, control resolution of inflammation, regulate host defense, and promote tolerance to prevent autoimmunity. Tumors and chronic pathogens can exploit these pathways to evade eradication by the immune system. Advances in understanding B7:CD28 pathways have ushered in a new era of immunotherapy with effective drugs to treat cancer, autoimmune diseases, infectious diseases, and transplant rejection. Here, we discuss current understanding of the mechanisms underlying the coinhibitory functions of CTLA-4, PD-1, PD-L1:B7-1 and PD-L2:RGMb interactions and less studied B7 family members, including HHLA2, VISTA, BTNL2, and BTN3A1, as well as their overlapping and unique roles in regulating immune responses, and the therapeutic potential of these insights. CI - Copyright © 2024 Elsevier Inc. All rights reserved. FAU - Burke, Kelly P AU - Burke KP AD - Department of Medical Oncology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA 02115, USA; Department of Immunology, Harvard Medical School, Boston, MA 02115, USA. FAU - Chaudhri, Apoorvi AU - Chaudhri A AD - Department of Medical Oncology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA 02115, USA. FAU - Freeman, Gordon J AU - Freeman GJ AD - Department of Medical Oncology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA 02115, USA. FAU - Sharpe, Arlene H AU - Sharpe AH AD - Department of Immunology, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA, USA; Brigham and Women's Hospital, Boston, MA 02115, USA. Electronic address: arlene_sharpe@hms.harvard.edu. LA - eng GR - P01 CA236749/CA/NCI NIH HHS/United States GR - P01 AI056299/AI/NIAID NIH HHS/United States GR - P50 CA101942/CA/NCI NIH HHS/United States GR - P01 AI108545/AI/NIAID NIH HHS/United States GR - P50 CA127003/CA/NCI NIH HHS/United States PT - Journal Article PT - Review PL - United States TA - Immunity JT - Immunity JID - 9432918 RN - 0 (CD28 Antigens) RN - 0 (CTLA-4 Antigen) RN - 0 (B7-1 Antigen) RN - 0 (HHLA2 protein, human) RN - 0 (Immunoglobulins) RN - 0 (BTNL2 protein, human) RN - 0 (Butyrophilins) RN - 0 (BTN3A1 protein, human) RN - 0 (Antigens, CD) SB - IM MH - Humans MH - *CD28 Antigens/metabolism MH - Friends MH - T-Lymphocytes MH - CTLA-4 Antigen/metabolism MH - Immunotherapy MH - *Autoimmune Diseases MH - B7-1 Antigen/metabolism MH - Immunoglobulins/metabolism MH - Butyrophilins/metabolism MH - Antigens, CD/metabolism PMC - PMC10889489 MID - NIHMS1961890 COIS- Declaration of interests A.H.S. has patents/pending royalties on the PD-1 pathway from Roche and Novartis. A.H.S. is on advisory boards for Elpiscience, Bicara, Monopteros, Fibrogen, Alixia, IOME, Corner Therapeutics, Bioentre, GlaxoSmithKline, Amgen, and Janssen. She also is on scientific advisory boards for the Massachusetts General Cancer Center, Program in Cellular and Molecular Medicine at Boston Children’s Hospital, the Human Oncology and Pathogenesis Program at Memorial Sloan Kettering Cancer Center, the Johns Hopkins Bloomberg Kimmel Institute for Cancer Immunotherapy, and the Gladstone Institute and is an academic editor for the Journal of Experimental Medicine. A.H.S. has funding from Quark/Iome; AbbVie; Calico; and Taiwan Bio. G.J.F. has patents/pending royalties on the PD-L1/PD-1 pathway from Roche, Merck MSD, Bristol-Myers-Squibb, Merck KGA, Boehringer-Ingelheim, AstraZeneca, Dako, Leica, Mayo Clinic, Eli Lilly, and Novartis. G.J.F. has patents or patent applications on the use of PD-L1, PD-L2, PD-1, RGMb, HHLA2, KIR3DL3, and BTNL2 in cancer immunotherapy. G.J.F. has served on advisory boards for iTeos, NextPoint, IgM, GV20, IOME, Bioentre, Santa Ana Bio, Simcere of America, and Geode. G.J.F. has equity in Nextpoint, Triursus, Xios, iTeos, IgM, Trillium, Invaria, GV20, Bioentre, and Geode. EDAT- 2024/02/15 00:42 MHDA- 2024/02/16 06:42 PMCR- 2025/02/13 CRDT- 2024/02/14 18:18 PHST- 2023/11/21 00:00 [received] PHST- 2024/01/17 00:00 [revised] PHST- 2024/01/17 00:00 [accepted] PHST- 2025/02/13 00:00 [pmc-release] PHST- 2024/02/16 06:42 [medline] PHST- 2024/02/15 00:42 [pubmed] PHST- 2024/02/14 18:18 [entrez] AID - S1074-7613(24)00038-4 [pii] AID - 10.1016/j.immuni.2024.01.013 [doi] PST - ppublish SO - Immunity. 2024 Feb 13;57(2):223-244. doi: 10.1016/j.immuni.2024.01.013.