fc0891d6e624f1cbe9107366c6581481 medsci-14-00465-v2.pdf 32a7af4d68246a61b73186b41790c524537eebdb medsci-14-00465-v2.pdf 009565ab080a851dafd7bdeefe1ab2c9ebf234de4cef0fe27556927842a591aa medsci-14-00465-v2.pdf Title: Early Identification of a Cytokine-Low Immune Phenotype in Suspected Sepsis Using a Point-of-Care Whole-Blood TNF- Release Assay Subject: Background/Objectives: Routine biomarkers and clinical scores do not directly assess functional immune responsiveness in suspected sepsis. We evaluated whether the TNF- Release Assay (TARA), a whole-blood LPS-induced functional assay, is associated with an early cytokine-low immune phenotype in Emergency Department (ED) patients. Methods: In this prospective single-center cohort, adults with suspected sepsis and NEWS2 3 were sampled at presentation, at 4 h and, when available, at 24 h. Immune phenotypes were defined independently of TARA by unsupervised k-means clustering of 12 circulating cytokines at 4 h, and a continuous Low-Response Cytokine Score (LRCS) was derived, with higher values indicating lower global cytokine concentrations. Results: Among 152 patients, three phenotypes were identified: C1 cytokine-low (n = 56), C2 inflammatory/intermediate (n = 63), and C3 IFN-/MCP-3/IL-17A-high (n = 33). Indicative TARA low-response status was independently associated with a higher LRCS after adjustment for clinical covariates and procalcitonin (PCT), both at presentation ( = 0.33; 95% CI, 0.14–0.53; p = 0.001) and at 4 h ( = 0.38; 95% CI, 0.19–0.57; p < 0.001). Indicative TARA low-response results at 4 h were most frequent in C1 and least frequent in C3 (75% vs. 45%). Exploratory clinical comparisons showed lower recorded early escalation in C1, whereas Sepsis-3 final diagnosis and in-hospital mortality were similar across phenotypes. Conclusions: A TARA low-response status was independently associated with an early circulating cytokine-low phenotype defined without reference to TARA or outcomes, providing information complementary to routine biomarkers and clinical severity scores. These exploratory findings support TARA as a complementary functional immune readout but require external validation and clinical-impact evaluation. Keywords: suspected sepsis; sepsis; host response; cytokine-low phenotype; TARA; TNF- release assay; innate immune hyporesponsiveness; emergency department Author: Erika P. Plata-Menchaca, Berta Cisteró, Adrian Ceccato, Veronica Monforte, Queralt Caus-Capdevila, Aina Areny-Balaguero, Elena Campaña-Duel, Marta Camprubí-Rimblas, Gemma Goma-Fernandez, Carla Guijarro, Patricia Salom, Monica Lopez, Juan Tajan, Tiago Teles De Castro, Vanessa Roman, Joan Vieyra, Judit Cubedo, Eduard Guerrero, Emili Gené, Antonio Artigas and Enrique Hernández-Jiménez Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Mon Aug 10 07:53:32 2026 CEST ModDate: Mon Aug 10 07:58:29 2026 CEST Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 19 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 723876 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- DORTIW+EURM10 Type 1 Builtin yes yes yes 146 0 ZDEJCT+URWPalladioL-Bold Type 1 Custom yes yes yes 147 0 MLTPMG+VnURWPalladioL Type 1 Custom yes yes yes 148 0 NTZLIT+URWPalladioL-Roma Type 1 Custom yes yes yes 149 0 QVIKRN+URWPalladioL-Ital Type 1 Custom yes yes yes 150 0 RQPVQI+CMSY10 Type 1 Builtin yes yes yes 151 0 CZOINJ+EURB10 Type 1 Builtin yes yes yes 152 0 FAECIJ+URWPalladioL-BoldItal Type 1 Custom yes yes yes 278 0 OOFKFM+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 265 0 OOFKEK+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 266 0 OOFKEL+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 267 0 OOFKFM+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 324 0 OOFKEK+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 325 0 OOFKEL+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 326 0 OOFKFM+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 344 0 OOFKEK+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 345 0 OOFKEL+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 346 0 OOFKFM+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 385 0 OOFKEK+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 386 0 OOFKEL+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 387 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-09-30 03:56:39 CEST RepresentationInformation: medsci-14-00465-v2.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-09-29 14:53:17 CEST Size: 723876 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 646 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Materials and Methods Children: Item: Title: Study Design, Setting and Ethics Item: Title: Participants, Clinical Data and Sampling Time Points Item: Title: TARA Functional Immune Assay Children: Item: Title: Plasma Cytokine Measurement Item: Title: Cytokine-Derived Immune Phenotype Definition Item: Title: Primary and Secondary Analyses Children: Item: Title: Clinical, Microbiological and Outcome Analyses Item: Title: Statistical Analysis Item: Title: Results Children: Item: Title: Study Population and Definition of Early Cytokine-Derived Immune Phenotypes Item: Title: TARA Low-Response Status Is Independently Associated with the Early Cytokine-Low Phenotype Item: Title: Cytokine-Derived Immune Phenotypes Show Longitudinal Biological Coherence Item: Title: Routine Biomarkers, Severity Scores and Hematological Variables Do Not Fully Reproduce the Cytokine-Low Phenotype Item: Title: Clinical Presentation, Microbiological Context and Exploratory Hospital Outcomes Across Immune Phenotypes Item: Title: Discussion Children: Item: Title: Biological Interpretation and Established Endotypes Item: Title: Microbiological Context and Longitudinal Interpretation Item: Title: Potential Clinical Implications Item: Title: Strengths and Limitations Item: Title: Conclusions Item: Title: References Info: Title: Early Identification of a Cytokine-Low Immune Phenotype in Suspected Sepsis Using a Point-of-Care Whole-Blood TNF- Release Assay Author: Erika P. Plata-Menchaca, Berta Cisteró, Adrian Ceccato, Veronica Monforte, Queralt Caus-Capdevila, Aina Areny-Balaguero, Elena Campaña-Duel, Marta Camprubí-Rimblas, Gemma Goma-Fernandez, Carla Guijarro, Patricia Salom, Monica Lopez, Juan Tajan, Tiago Teles De Castro, Vanessa Roman, Joan Vieyra, Judit Cubedo, Eduard Guerrero, Emili Gené, Antonio Artigas and Enrique Hernández-Jiménez Subject: Background/Objectives: Routine biomarkers and clinical scores do not directly assess functional immune responsiveness in suspected sepsis. We evaluated whether the TNF- Release Assay (TARA), a whole-blood LPS-induced functional assay, is associated with an early cytokine-low immune phenotype in Emergency Department (ED) patients. Methods: In this prospective single-center cohort, adults with suspected sepsis and NEWS2 3 were sampled at presentation, at 4 h and, when available, at 24 h. Immune phenotypes were defined independently of TARA by unsupervised k-means clustering of 12 circulating cytokines at 4 h, and a continuous Low-Response Cytokine Score (LRCS) was derived, with higher values indicating lower global cytokine concentrations. Results: Among 152 patients, three phenotypes were identified: C1 cytokine-low (n = 56), C2 inflammatory/intermediate (n = 63), and C3 IFN-/MCP-3/IL-17A-high (n = 33). Indicative TARA low-response status was independently associated with a higher LRCS after adjustment for clinical covariates and procalcitonin (PCT), both at presentation ( = 0.33; 95% CI, 0.14–0.53; p = 0.001) and at 4 h ( = 0.38; 95% CI, 0.19–0.57; p < 0.001). Indicative TARA low-response results at 4 h were most frequent in C1 and least frequent in C3 (75% vs. 45%). Exploratory clinical comparisons showed lower recorded early escalation in C1, whereas Sepsis-3 final diagnosis and in-hospital mortality were similar across phenotypes. Conclusions: A TARA low-response status was independently associated with an early circulating cytokine-low phenotype defined without reference to TARA or outcomes, providing information complementary to routine biomarkers and clinical severity scores. These exploratory findings support TARA as a complementary functional immune readout but require external validation and clinical-impact evaluation. 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