{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T16:17:25Z","timestamp":1777738645632,"version":"3.51.4"},"reference-count":52,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2022,12,11]],"date-time":"2022-12-11T00:00:00Z","timestamp":1670716800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institute of Allergy","doi-asserted-by":"publisher","award":["R01AI132348"],"award-info":[{"award-number":["R01AI132348"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institute of Allergy","doi-asserted-by":"publisher","award":["75N93019C00052"],"award-info":[{"award-number":["75N93019C00052"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Center for Influenza Vaccine Research for High Risk Populations (CIVR-HRP)","award":["R01AI132348"],"award-info":[{"award-number":["R01AI132348"]}]},{"name":"Center for Influenza Vaccine Research for High Risk Populations (CIVR-HRP)","award":["75N93019C00052"],"award-info":[{"award-number":["75N93019C00052"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Older adults experience declining influenza vaccine-induced immunity and are at higher risk of influenza and its complications. For this reason, high dose (e.g., Fluzone) and adjuvanted (e.g., Fluad) vaccines are preferentially recommended for people age 65 years and older. However, T cell transcriptional activity shaping the humoral immune responses to Fluzone and Fluad vaccines in older adults is still poorly understood. We designed a study of 234 older adults (\u226565 years old) who were randomly allocated to receive Fluzone or Fluad vaccine and provided blood samples at baseline and at Day 28 after immunization. We measured the humoral immune responses (hemagglutination inhibition\/HAI antibody titer) to influenza A\/H3N2 and performed mRNA-Seq transcriptional profiling in purified CD4+ T cells, in order to identify T cell signatures that might explain differences in humoral immune response by vaccine type. Given the large differences in formulation (higher antigen dose vs adjuvant), our hypothesis was that each vaccine elicited a distinct transcriptomic response after vaccination. Thus, the main focus of our study was to identify the differential gene expression influencing the antibody titer in the two vaccine groups. Our analyses identified three differentially expressed, functionally linked genes\/proteins in CD4+ T cells: the calcium\/calmodulin dependent serine\/threonine kinase IV (CaMKIV); its regulator the TMEM38B\/transmembrane protein 38B, involved in maintenance of intracellular Ca2+ release; and the transcriptional coactivator CBP\/CREB binding protein, as regulators of transcriptional activity\/function in CD4+ T cells that impact differences in immune response by vaccine type. Significantly enriched T cell-specific pathways\/biological processes were also identified that point to the importance of genes\/proteins involved in Th1\/Th2 cell differentiation, IL-17 signaling, calcium signaling, Notch signaling, MAPK signaling, and regulation of TRP cation Ca2+ channels in humoral immunity after influenza vaccination. In summary, we identified the genes\/proteins and pathways essential for cell activation and function in CD4+ T cells that are associated with differences in influenza vaccine-induced humoral immunity by vaccine type. These findings provide an additional mechanistic perspective for achieving protective immunity in older adults.<\/jats:p>","DOI":"10.3390\/v14122763","type":"journal-article","created":{"date-parts":[[2022,12,12]],"date-time":"2022-12-12T03:36:09Z","timestamp":1670816169000},"page":"2763","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["T Cell Transcriptional Signatures of Influenza A\/H3N2 Antibody Response to High Dose Influenza and Adjuvanted Influenza Vaccine in Older Adults"],"prefix":"10.3390","volume":"14","author":[{"given":"Iana H.","family":"Haralambieva","sequence":"first","affiliation":[{"name":"Mayo Clinic Vaccine Research Group, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huy Quang","family":"Quach","sequence":"additional","affiliation":[{"name":"Mayo Clinic Vaccine Research Group, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Inna G.","family":"Ovsyannikova","sequence":"additional","affiliation":[{"name":"Mayo Clinic Vaccine Research Group, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Krista M.","family":"Goergen","sequence":"additional","affiliation":[{"name":"Department of Quantitative Health Sciences, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Diane E.","family":"Grill","sequence":"additional","affiliation":[{"name":"Department of Quantitative Health Sciences, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gregory A.","family":"Poland","sequence":"additional","affiliation":[{"name":"Mayo Clinic Vaccine Research Group, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6164-4967","authenticated-orcid":false,"given":"Richard B.","family":"Kennedy","sequence":"additional","affiliation":[{"name":"Mayo Clinic Vaccine Research Group, Mayo Clinic, Rochester, MN 55905, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,11]]},"reference":[{"key":"ref_1","unstructured":"(2022, December 05). Available online: https:\/\/www.cdc.gov\/flu\/about\/burden\/index.html."},{"key":"ref_2","unstructured":"(2010). Estimates of deaths associated with seasonal influenza\u2014United States, 1976\u20132007. MMWR Morb. Mortal. Wkly. Rep., 59, 1057\u20131062."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Reed, C., Chaves, S.S., Daily Kirley, P., Emerson, R., Aragon, D., Hancock, E.B., Butler, L., Baumbach, J., Hollick, G., and Bennett, N.M. (2015). Estimating influenza disease burden from population-based surveillance data in the United States. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0118369"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1427","DOI":"10.1093\/cid\/cis211","article-title":"Hospitalizations associated with influenza and respiratory syncytial virus in the United States, 1993\u20132008","volume":"54","author":"Zhou","year":"2012","journal-title":"Clin. Infect. Dis."},{"key":"ref_5","unstructured":"(2022, December 05). Available online: https:\/\/www.cdc.gov\/flu\/highrisk\/65over.htm."},{"key":"ref_6","unstructured":"World Health Organization (2019). Global Influenza Strategy 2019\u20132030, World Health Organization."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1016\/j.vaccine.2005.08.105","article-title":"Antibody response to influenza vaccination in the elderly: A quantitative review","volume":"24","author":"Goodwin","year":"2006","journal-title":"Vaccine"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Haralambieva, I.H., Painter, S.D., Kennedy, R.B., Ovsyannikova, I.G., Lambert, N.D., Goergen, K.M., Oberg, A.L., and Poland, G.A. (2015). The impact of immunosenescence on humoral immune response variation after influenza A\/H1N1 vaccination in older subjects. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0122282"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1056\/NEJMoa1315727","article-title":"Efficacy of high-dose versus standard-dose influenza vaccine in older adults","volume":"371","author":"DiazGranados","year":"2014","journal-title":"N. Engl. J. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1016\/j.vaccine.2016.12.011","article-title":"Effectiveness of MF59-adjuvanted seasonal influenza vaccine in the elderly: A systematic review and meta-analysis","volume":"35","author":"Domnich","year":"2017","journal-title":"Vaccine"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5027","DOI":"10.1016\/j.vaccine.2014.07.013","article-title":"Comparison of the safety and immunogenicity of an MF59\u00ae-adjuvanted with a non-adjuvanted seasonal influenza vaccine in elderly subjects","volume":"32","author":"Frey","year":"2014","journal-title":"Vaccine"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1016\/S2213-2600(17)30235-7","article-title":"Comparative effectiveness of high-dose versus standard-dose influenza vaccination on numbers of US nursing home residents admitted to hospital: A cluster-randomised trial","volume":"5","author":"Gravenstein","year":"2017","journal-title":"Lancet Respir. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1080\/14760584.2019.1622418","article-title":"Adjuvanted versus nonadjuvanted influenza vaccines and risk of hospitalizations for pneumonia and cerebro\/cardiovascular events in the elderly","volume":"18","author":"Lapi","year":"2019","journal-title":"Expert Rev. Vaccines"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e4237","DOI":"10.1093\/cid\/ciaa1233","article-title":"Cluster-randomized Trial of Adjuvanted Versus Nonadjuvanted Trivalent Influenza Vaccine in 823 US Nursing Homes","volume":"73","author":"McConeghy","year":"2021","journal-title":"Clin. Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1093\/cid\/ciab152","article-title":"Relative Effectiveness of Adjuvanted Trivalent Inactivated Influenza Vaccine Versus Egg-derived Quadrivalent Inactivated Influenza Vaccines and High-dose Trivalent Influenza Vaccine in Preventing Influenza-related Medical Encounters in US Adults \u2265 65 Years During the 2017\u20132018 and 2018\u20132019 Influenza Seasons","volume":"73","author":"Boikos","year":"2021","journal-title":"Clin. Infect. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1824","DOI":"10.1002\/eji.202049054","article-title":"Transcriptional signatures associated with rubella virus-specific humoral immunity after a third dose of MMR vaccine in women of childbearing age","volume":"51","author":"Haralambieva","year":"2021","journal-title":"Eur. J. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Kaufmann, L., Syedbasha, M., Vogt, D., Hollenstein, Y., Hartmann, J., Linnik, J.E., and Egli, A. (2017). An optimized hemagglutination inhibition (HI) assay to quantify influenza-specific antibody titers. J. Vis. Exp., 55833.","DOI":"10.3791\/55833-v"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1038\/gene.2016.34","article-title":"Gene signatures associated with adaptive humoral immunity following seasonal influenza A\/H1N1 vaccination","volume":"17","author":"Ovsyannikova","year":"2016","journal-title":"Genes Immun."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Quach, H.Q., Chen, J., Monroe, J.M., Ratishvili, T., Warner, N.D., Grill, D.E., Haralambieva, I.H., Ovsyannikova, I.G., Poland, G.A., and Kennedy, R.B. (2022). The influence of sex, BMI, and age on cellular and humoral immune responses against measles after a 3rd dose of MMR vaccine. J. Infect. Dis., jiac351.","DOI":"10.1093\/infdis\/jiac351"},{"key":"ref_20","unstructured":"Webster, R., Cox, N., and St\u00f6hr, K. (2002). WHO Animal Influenza Manual, World Health Organization. WHO\/CDS\/CSR\/NCS\/2002.5."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"445","DOI":"10.3389\/fimmu.2017.00445","article-title":"Integration of Immune Cell Populations, mRNA-Seq, and CpG Methylation to Better Predict Humoral Immunity to Influenza Vaccination: Dependence of mRNA-Seq\/CpG Methylation on Immune Cell Populations","volume":"8","author":"Zimmermann","year":"2017","journal-title":"Front. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3993","DOI":"10.1016\/j.vaccine.2016.06.034","article-title":"Transcriptional signatures of influenza A\/H1N1-specific IgG memory-like B cell response in older individuals","volume":"34","author":"Haralambieva","year":"2016","journal-title":"Vaccine"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Kalari, K.R., Nair, A.A., Bhavsar, J.D., O\u2019Brien, D.R., Davila, J.I., Bockol, M.A., Nie, J., Tang, X., Baheti, S., and Doughty, J.B. (2014). MAP-RSeq: Mayo analysis pipeline for RNA sequencing. BMC Bioinform., 15.","DOI":"10.1186\/1471-2105-15-224"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1093\/biostatistics\/kxr054","article-title":"Removing technical variability in RNA-seq data using conditional quantile normalization","volume":"13","author":"Hansen","year":"2012","journal-title":"Biostatistics"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1007\/978-3-642-04898-2_248","article-title":"False Discovery Rate","volume":"1","author":"Storey","year":"2011","journal-title":"Int. Encycl. Stat. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"clusterProfiler: An R package for comparing biological themes among gene clusters","volume":"16","author":"Yu","year":"2012","journal-title":"OMICS J. Integr. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1111\/j.2517-6161.1995.tb02031.x","article-title":"Controlling the false discovery rate: A practical and powerful approach to multiple testing","volume":"57","author":"Benjamini","year":"1995","journal-title":"J. R. Stat. Soc. Ser. B"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"180","DOI":"10.3389\/fimmu.2019.00180","article-title":"Sex differences in older adults\u2019 immune responses to seasonal influenza vaccination","volume":"10","author":"Voigt","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.apsb.2019.12.006","article-title":"Solute carrier transporters: The metabolic gatekeepers of immune cells","volume":"10","author":"Song","year":"2020","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"786","DOI":"10.1038\/ni.2067","article-title":"Systems biology of vaccination for seasonal influenza in humans","volume":"12","author":"Nakaya","year":"2011","journal-title":"Nat. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1016\/j.smim.2013.05.003","article-title":"Systems biological approaches to measure and understand vaccine immunity in humans","volume":"25","author":"Li","year":"2013","journal-title":"Semin. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"12300","DOI":"10.1073\/pnas.1400476111","article-title":"Systems vaccinology: Probing humanity\u2019s diverse immune systems with vaccines","volume":"111","author":"Pulendran","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1505","DOI":"10.1126\/science.281.5382.1505","article-title":"CBP: A signal-regulated transcriptional coactivator controlled by nuclear calcium and CaM kinase IV","volume":"281","author":"Chawla","year":"1998","journal-title":"Science"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"7977","DOI":"10.1074\/jbc.M808620200","article-title":"Ribosomal protein S19 interacts with macrophage migration inhibitory factor and attenuates its pro-inflammatory function","volume":"284","author":"Filip","year":"2009","journal-title":"J. Biol. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3509","DOI":"10.1002\/eji.200737357","article-title":"Regulation of constitutive and microbial pathogen-induced human macrophage migration inhibitory factor (MIF) gene expression","volume":"37","author":"Roger","year":"2007","journal-title":"Eur. J. Immunol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2419","DOI":"10.3389\/fimmu.2019.02419","article-title":"NLRX1 Is a Multifaceted and Enigmatic Regulator of Immune System Function","volume":"10","author":"Morrison","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1093\/nar\/28.1.27","article-title":"KEGG: Kyoto encyclopedia of genes and genomes","volume":"28","author":"Kanehisa","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1128\/MCB.22.4.1001-1015.2002","article-title":"Rap1 functions as a key regulator of T-cell and antigen-presenting cell interactions and modulates T-cell responses","volume":"22","author":"Katagiri","year":"2002","journal-title":"Mol. Cell. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1038\/s41598-017-17735-x","article-title":"Transcriptomic signatures of cellular and humoral immune responses in older adults after seasonal influenza vaccination identified by data-driven clustering","volume":"8","author":"Voigt","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1853","DOI":"10.1073\/pnas.1519690113","article-title":"Systems biology of immunity to MF59-adjuvanted versus nonadjuvanted trivalent seasonal influenza vaccines in early childhood","volume":"113","author":"Nakaya","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"6413","DOI":"10.4049\/jimmunol.1001829","article-title":"The role of the transcription factor CREB in immune function","volume":"185","author":"Wen","year":"2010","journal-title":"J. Immunol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1038\/379081a0","article-title":"Defective thymocyte proliferation and IL-2 production in transgenic mice expressing a dominant-negative form of CREB","volume":"379","author":"Barton","year":"1996","journal-title":"Nature"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1762","DOI":"10.4049\/jimmunol.165.4.1762","article-title":"Defective Th function induced by a dominant-negative cAMP response element binding protein mutation is reversed by Bcl-2","volume":"165","author":"Zhang","year":"2000","journal-title":"J. Immunol."},{"key":"ref_44","first-page":"8524","article-title":"CaMK4 Controls the Humoral Response via the T Follicular Helper Compartment and Regulates Autoimmunity","volume":"74","author":"Scherlinger","year":"2022","journal-title":"Arthritis Rheumatol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2583","DOI":"10.1002\/eji.200636606","article-title":"MSK regulate TCR-induced CREB phosphorylation but not immediate early gene transcription","volume":"37","author":"Kaiser","year":"2007","journal-title":"Eur. J. Immunol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"284","DOI":"10.4049\/jimmunol.166.1.284","article-title":"Multiple signals required for cyclic AMP-responsive element binding protein (CREB) binding protein interaction induced by CD3\/CD28 costimulation","volume":"166","author":"Yu","year":"2001","journal-title":"J. Immunol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1016\/S0896-6273(00)80738-2","article-title":"Regulation of CBP-mediated transcription by neuronal calcium signaling","volume":"22","author":"Hu","year":"1999","journal-title":"Neuron"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1128\/MCB.22.1.23-29.2002","article-title":"Defective signaling in a subpopulation of CD4+ T cells in the absence of Ca2+\/calmodulin-dependent protein kinase IV","volume":"22","author":"Anderson","year":"2002","journal-title":"Mol. Cell. Biol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1182\/blood-2007-05-091173","article-title":"Calmodulin-dependent kinase IV links Toll-like receptor 4 signaling with survival pathway of activated dendritic cells","volume":"111","author":"Illario","year":"2008","journal-title":"Blood"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"4302","DOI":"10.1523\/JNEUROSCI.17-11-04302.1997","article-title":"The fusiform face area: A module in human extrastriate cortex specialized for face perception","volume":"17","author":"Kanwisher","year":"1997","journal-title":"J. Neurosci."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1038\/s41577-018-0110-7","article-title":"Calcium signalling in T cells","volume":"19","author":"Trebak","year":"2019","journal-title":"Nat. Rev. Immunol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"21095","DOI":"10.1073\/pnas.1319784110","article-title":"The adjuvant MF59 induces ATP release from muscle that potentiates response to vaccination","volume":"110","author":"Vono","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/12\/2763\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:39:27Z","timestamp":1760146767000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/12\/2763"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,11]]},"references-count":52,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["v14122763"],"URL":"https:\/\/doi.org\/10.3390\/v14122763","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,11]]}}}