Comparison of whole blood cytokine immunoassays for rapid, functional immune phenotyping in critically ill patients with sepsis
Abstract Background Sepsis is characterized by highly heterogeneous immune responses associated with a spectrum of disease severity. Methods that rapidly and sensitively profile these immune responses can potentially personalize immune-adjuvant therapies for sepsis. We hypothesized that the ELLA mic...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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SpringerOpen
2023-10-01
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| Series: | Intensive Care Medicine Experimental |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40635-023-00556-w |
| _version_ | 1797578601465380864 |
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| author | Anthony S. Bonavia Abigail Samuelsen Menglu Liang Jodi Hanson Daniel McKeone Zissis C. Chroneos E. Scott Halstead |
| author_facet | Anthony S. Bonavia Abigail Samuelsen Menglu Liang Jodi Hanson Daniel McKeone Zissis C. Chroneos E. Scott Halstead |
| author_sort | Anthony S. Bonavia |
| collection | DOAJ |
| description | Abstract Background Sepsis is characterized by highly heterogeneous immune responses associated with a spectrum of disease severity. Methods that rapidly and sensitively profile these immune responses can potentially personalize immune-adjuvant therapies for sepsis. We hypothesized that the ELLA microfluidic approach to measure cytokine production from the whole blood of septic and critically ill patients would deliver faster, more precise results than the existing optic-driven ELISpot quantification. We tested our hypothesis by measuring ex vivo-stimulated production of TNF and IFNγ in critically ill and septic patients (n = 22), critically ill and non-septic patients (n = 10), and healthy volunteers (n = 10) through both ELLA and ELISpot immunoassays. Blood samples were subjected to one of three stimulants for 4 h or 18 h durations during days 1, 7–10, and 14 of critical illness. Stimulants for lymphocytes included anti-CD3/anti-CD28 and phorbol 12-myristate 13-acetate (PMA), whereas LPS was used for monocytes. Stimulated TNF and IFNγ concentrations were then associated with 30-day mortality. Results Both ELISpot and ELLA immunoassays showed substantial agreement in TNF concentrations post 4 h and 18 h LPS stimulation, with concordance correlation coefficients at 0.62 and 0.60, respectively. ELLA had a broad dynamic measurement range and provided accurate TNF and IFNγ readings at both minimal and elevated cytokine concentrations (with mean coefficients of variation between triplicate readings at 2.1 ± 1.4% and 4.9 ± 7.2%, respectively). However, there was no association between the ELLA-determined cytokine concentrations on the first day of critical illness and 30-day mortality rate. In contrast, using the ELISpot for cytokine quantification revealed that non-survivors had reduced baseline TNF levels at 18 h, decreased LPS-induced TNF levels at 18 h, and diminished TNF levels post 4 h/18 h anti-CD3/28 stimulation. Conclusions Our study affirms the feasibility of obtaining dependable immune phenotyping data within 6 h of blood collection from critically ill patients, both septic and non-septic, using the ELLA immunoassay. Both ELLA and ELISpot can offer valuable insights into prognosis, therapeutic strategies, and the underlying mechanisms of sepsis development. |
| first_indexed | 2024-03-10T22:24:07Z |
| format | Article |
| id | doaj.art-6b7f80b19ab24f1fbce8e70d047f84d9 |
| institution | Directory Open Access Journal |
| issn | 2197-425X |
| language | English |
| last_indexed | 2024-03-10T22:24:07Z |
| publishDate | 2023-10-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Intensive Care Medicine Experimental |
| spelling | doaj.art-6b7f80b19ab24f1fbce8e70d047f84d92023-11-19T12:10:12ZengSpringerOpenIntensive Care Medicine Experimental2197-425X2023-10-0111111810.1186/s40635-023-00556-wComparison of whole blood cytokine immunoassays for rapid, functional immune phenotyping in critically ill patients with sepsisAnthony S. Bonavia0Abigail Samuelsen1Menglu Liang2Jodi Hanson3Daniel McKeone4Zissis C. Chroneos5E. Scott Halstead6Department of Anesthesiology and Perioperative Medicine, Penn State Milton S. Hershey Medical CenterDepartment of Anesthesiology and Perioperative Medicine, Penn State Milton S. Hershey Medical CenterDepartment of Epidemiology and Biostatistics, School of Public Health, University of MarylandCellular TechnologyDepartment of Pediatrics, Penn State Milton S. Hershey Medical CenterDepartment of Pediatrics, Penn State Milton S. Hershey Medical CenterDepartment of Pediatrics, Penn State Milton S. Hershey Medical CenterAbstract Background Sepsis is characterized by highly heterogeneous immune responses associated with a spectrum of disease severity. Methods that rapidly and sensitively profile these immune responses can potentially personalize immune-adjuvant therapies for sepsis. We hypothesized that the ELLA microfluidic approach to measure cytokine production from the whole blood of septic and critically ill patients would deliver faster, more precise results than the existing optic-driven ELISpot quantification. We tested our hypothesis by measuring ex vivo-stimulated production of TNF and IFNγ in critically ill and septic patients (n = 22), critically ill and non-septic patients (n = 10), and healthy volunteers (n = 10) through both ELLA and ELISpot immunoassays. Blood samples were subjected to one of three stimulants for 4 h or 18 h durations during days 1, 7–10, and 14 of critical illness. Stimulants for lymphocytes included anti-CD3/anti-CD28 and phorbol 12-myristate 13-acetate (PMA), whereas LPS was used for monocytes. Stimulated TNF and IFNγ concentrations were then associated with 30-day mortality. Results Both ELISpot and ELLA immunoassays showed substantial agreement in TNF concentrations post 4 h and 18 h LPS stimulation, with concordance correlation coefficients at 0.62 and 0.60, respectively. ELLA had a broad dynamic measurement range and provided accurate TNF and IFNγ readings at both minimal and elevated cytokine concentrations (with mean coefficients of variation between triplicate readings at 2.1 ± 1.4% and 4.9 ± 7.2%, respectively). However, there was no association between the ELLA-determined cytokine concentrations on the first day of critical illness and 30-day mortality rate. In contrast, using the ELISpot for cytokine quantification revealed that non-survivors had reduced baseline TNF levels at 18 h, decreased LPS-induced TNF levels at 18 h, and diminished TNF levels post 4 h/18 h anti-CD3/28 stimulation. Conclusions Our study affirms the feasibility of obtaining dependable immune phenotyping data within 6 h of blood collection from critically ill patients, both septic and non-septic, using the ELLA immunoassay. Both ELLA and ELISpot can offer valuable insights into prognosis, therapeutic strategies, and the underlying mechanisms of sepsis development.https://doi.org/10.1186/s40635-023-00556-wSepsisImmunoassayCytokinesCritical IllnessHumans |
| spellingShingle | Anthony S. Bonavia Abigail Samuelsen Menglu Liang Jodi Hanson Daniel McKeone Zissis C. Chroneos E. Scott Halstead Comparison of whole blood cytokine immunoassays for rapid, functional immune phenotyping in critically ill patients with sepsis Intensive Care Medicine Experimental Sepsis Immunoassay Cytokines Critical Illness Humans |
| title | Comparison of whole blood cytokine immunoassays for rapid, functional immune phenotyping in critically ill patients with sepsis |
| title_full | Comparison of whole blood cytokine immunoassays for rapid, functional immune phenotyping in critically ill patients with sepsis |
| title_fullStr | Comparison of whole blood cytokine immunoassays for rapid, functional immune phenotyping in critically ill patients with sepsis |
| title_full_unstemmed | Comparison of whole blood cytokine immunoassays for rapid, functional immune phenotyping in critically ill patients with sepsis |
| title_short | Comparison of whole blood cytokine immunoassays for rapid, functional immune phenotyping in critically ill patients with sepsis |
| title_sort | comparison of whole blood cytokine immunoassays for rapid functional immune phenotyping in critically ill patients with sepsis |
| topic | Sepsis Immunoassay Cytokines Critical Illness Humans |
| url | https://doi.org/10.1186/s40635-023-00556-w |
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