From: Blood-based biomarkers of inflammation in amyotrophic lateral sclerosis
Measured biomarker(s) | Tissue and detection method | Mean disease duration at sample donation | Number of samples | Potential Diagnostic Value | Study Reference Clinical Trial ID |
---|---|---|---|---|---|
Cell populations | |||||
 Monocyte subpopulations | Monocyte isolation kit from peripheral blood, and flow cytometry | Not reported | n = 68 ALS (1st cohort) n = 55 controls (1st cohort) n = 100 ALS (2nd cohort) n = 60 controls (2nd cohort) | - CD14−/low/CD16+ monocytes decreased in ALS | Beers et al., 2020 [26] |
 Subpopulations of T cells, B cells, natural killer cells, and antigen presenting cells | Peripheral blood and FACS | 2.48 years | n = 73 ALS n = 48 controls | - increased Th1 and Th17 cells in ALS - decreased Th2 and Treg cells in ALS - increased NK cells and monocytes in ALS | Jin et al., 2020 [27] |
 neutrophil-to-lymphocyte ratio (NLR) | From whole blood | Not reported | n = 80 ALS n = 80 matched controls(n = 41 ALS and n = 41 matched controls at follow-up 3-6 m after initial donation; n = 22 ALS patients and n = 22 matched controls at the third donation at least 3 months after the second) | - NLR was consistently elevated in ALS samples | Keizman et al., 2009 [28] |
Candidate-based soluble factor analysis | |||||
 IL-2, IL-6, IL-10, IFN-γ, and TNF | Plasma, BioPlex | 21.37 months (median) | n = 79 ALS n = 79 controls | - all measured cytokines were increased in ALS | Tortelli et al., 2020 [29] |
 CD14, LBP and CRP | Serum; ELISAs | Not reported | n = 68 ALS (1st cohort) n = 55 controls (1st cohort) n = 100 ALS (2nd cohort) n = 60 controls (2nd cohort) | - soluble CD14 increased in ALS (both cohorts) - LBP increased in ALS (both cohorts) - CRP increased in ALS (both cohorts) | Beers et al., 2020 [26] |
 IL-1b, IL-6, IL-10, IL-12, TNF,IFN-γ, IL17a, and IL-23 | Serum; ELISAs | 2.48 years | n = 73 ALS n = 48 controls | - increased IL-1b, IL-6, and IFN-γ in ALS - decreased IL-10 in ALS - no difference in TNF, IL-12, IL-17a, and IL-23 | Jin et al., 2020 [27] |
 CD5L, Ficolin-3 | Plasma; ELISAs | 739.9 months (median) | n = 37 ALS n = 30 controls | CD5L and Ficolin-3 are increased in ALS | Mohanty et al., 2020 [30] |
 IL-2, IL-1b, TNF, IFN-γ and IL-4 | Serum; ELISAs | 40.4 months | n = 35 ALS n = 30 controls | - increased IL-4 and IL-1b in ALS - decreased IFN-γ in ALS - no difference in IL-2 and TNF | Polverino et al., 2020 [31] |
 MCP-1, eotaxin-1, IL-18, TNF, CRP, IL-1, sTREM2 | Plasma; MSD assay | Not reported | n = 108 ALS n = 41 controls (n = 85 ALS with samples 2+ visits) | - MCP-1 and IL-18 are increased in ALS - sTREM2 is increased in ALS | Huang et a;., 2020 [32] NCT01495390 |
 IL-6 | Plasma; Chemokine assay | 25.7 months | n = 82 ALS n = 43 controls | Not upregulated in ALS (trend) | Pronto-Laborinho et al., 2019 [33] |
 IP-10, MCP-1, MIG, RANTES, IL-2, IL-4, IL-6, IL-8, IL-10, IL-17a, TNF, IGF-g, sTNFR1, sTNFR2 | Plasma; cytometric bead array and ELISA | 3 years | n = 68 ALS n = 62 controls over the first time point. n = 24 ALS at the second time point (6–12 months later) | IL-6 + IL-8: upregulated in ALS IL-2 (low) and IL-6 (high) predict ALS diagnosis | Prado et al., 2018 [34] |
 Gene expression of 45 genes | Serum; individual RT-qPCRs | Within the first 2 months of diagnosis | n = 22 sALS n = 13 controls | - ITGB2, INPP5D, SELL, ICAM1, MMP9 and TIMP2 are upregulated in ALS - CCL5, CXC5R, IL10, TGFB2, IL10RA, IL-6, CD2 and TRBC1 are downregulated in ALS | Andres-Benito et al., 2017 [35] |
 Gene expression of 37 brain-enriched and inflammation-associated microRNAs | Plasma; individual RT-qPCRs | 1.8 years | n = 50 ALS n = 50 FTD n = 50 AD n = 50 PD n = 50 controls | miR-206/miR-31 and miR-206/ miR-125b and miR-99/ miR-338-3p most effectively differentiate between ALS and control | Sheinerman et al., 2017 [36] |
 CC-16 | Plasma; ELISA | 27 months | n = 81 ALS n = 30 controls | Upregulated in ALS | Pronto-Laborinho et al., 2017 [37] |
 TNF, MCP-1, IL6, IL8, IL2, IFN-γ, IL1-beta, IL10, IL4, IL5, IL17, TNFR1, ELAM-1 | Plasma; different per individual dataset (25 studies) | different per individual dataset (25 studies) | pooled n = 812 ALS pooled n = 639 controls | TNF, TNFR-1, IL-6, IL-1β, and IL-8 levels were elevated in ALS. | Hu et al., 2017 |
 | [38] | ||||
 TNF, IL-8, IL-6, IL-10 | Serum; multiplex assay | Not reported | n = 19 ALS n = 10 controls | - IL-6 was increased in ALS. - IL-8 was increased in ALS. | Blasco et al., 2016 [39] |
 IL-1β, IL-18, IL-33, IL-37, IL-1Ra, sIL-1R2, IL-18BP, sIL-1R4 | Serum; individual ELISAs | 11.32 months | n = 144 sALS n = 40 controls | - IL-18 was increased in ALS. - IL-18BP was increased in ALS. | Italiani et al., 2014 [40] |
 IL-33, soluble ST2 | Serum; individual ELISAs | Not reported | n = 42 ALS n = 38 controls | - IL-33 was increased in ALS - soluble ST2 was decreased in ALS | Lin et al., 2012 [41] |
 IL-17A | Serum; individual ELISA | 23.4 months | n = 32 ALS n = 14 controls | IL-17A was increased in ALS. | Fiala et al., 2010 [42] |
 kynurenine pathway (tryptophan, picolinic acid) | Serum; HPLC, gas chromatography mass spectrometry | Not reported | n = 140 ALS n = 35 controls | - TRP and KYN is increased ALS - PIC is decreased in ALS | Chen et al., 2010 [43] |
 eotaxin, eotaxin-3, IL-8, IP-10, MCP-1, MCP-4, MDC, MIP-1b, TARC | Serum; solid-phase sandwich immuno-assay | Not reported | n = 20 ALS n = 20 controls (non-ALS neurologic) | No differences. | Kuhle et al., 2009 [44] |
 TNF, IFN-γ, and NO | Serum; individual ELISAs and NO by determining nitrite and nitrate levels | 12 months | n = 22 ALS n = 20 controls | TNF, IFN-γ, and NO were all increased in ALS | Babu et al., 2008 [45] |
 RANTES | Serum; individual ELISA | Not reported | n = 20 ALS n = 14 NIND n = 13 controls | RANTES was increased in ALS serum | Rentzos et al., 2008 [46] |
 MCP-1 | Serum; individual ELISA | 19.4 months | n = 27 ALS n = 30 NIND | MCP-1 was increased in ALS | Baron et al., 2005 [47] |
 MCP-1 | Serum; individual ELISA | 8 months (median) | n = 29 ALS n = 11 controls | MCP-1 was not altered in ALS | Wilms et al., 2005 [48] |
 wide-range C-reactive protein (wrCRP) | From whole blood | Not reported | n = 80 ALS n = 80 matched controls | - wrCRP was consistently elevated in ALS samples | Keizman et al., 2009 [28] |
Unbiased analyses of serum or plasma | |||||
 miRNA gene expression | Plasma; next generation sequencing on neural-enriched extracellular vesicles | Not reported | n = 10 + 10 (replication set) ALS n = 10 + 10 (replication set) controls | Eight miRNAs were differentially expressed between ALS samples and controls after replication. This included miR-146a-5p, which was upregulated in ALS samples and is associated with inflammation (monocytes) [49]. | Banack et al.;. 2020 [50]Some samples from NCT03580616. |
 Protein abundance detectable by mass spectrometry | Plasma; mass spectrometry | 747 months (median) | n = 42 ALS n = 18 controls | 30 proteins are differentially detected between ALS and controls. IPA analysis identified two networks of interacting proteins that differ between ALS and controls; IL-1 and NFkB. | Xu et al., 2018 [51] |
Blood-derived cells, in vitro assays | |||||
 Transcriptomic analysis | Gene expression of blood monocyte-derived macrophages, by RNAseq and RT-qPCR | Not reported | n = 5 controls n = 5 sALS n = 5 C9-ALS | - Increased type I interferon signature (pathway analysis) - increased gene expression of MX1, OASL, OAS2, IF44L | McCauley et al., 2020 [52] |
 Cell surface expression of VLA4, TLR4, CXCR3, CCR5, CXCR4, IFN-γ, CCR2, CD11B | Flow cytometry on blood-isolated T-cells, B-cells, monocytes, and NK cells | Not reported | n = 10 ALS n = 10 controls | - CXCR4, CXCR3, CCR2, CCR5 increased on ALS T-cells. - CD11B, CCR2 decreased on ALS monocytes - The combination of the analyzed markers could significantly predict the categorization into ALS or healthy donors, with CXCR3 and CCR5 on T cells comprising the strongest predictors. | Perner et al., 2018 [53] |
 Number of migrating cells | Boyden chamber. All cells migrated to the lower well after 2.5 h were stained using lineage antibody and counted by flow cytometry. | Not reported | n = 10 ALS n = 10 controls | More ALS CD45+ cells chemotaxis with IP-10 chemoattractant. | Perner et al., 2018 [53] |
 Frequency of myeloid dendritic cells | Flow cytometry (CD1chighCD19−) | Not calculated | n = 20 ALS n = 10 healthy donors | - Less circulating myeloid dendritic cells in ALS. - Increased CD62L expression on circulating myeloid dendritic cells in ALS. | Rusconi et al., 2017 [54] |
 Concentrations of TNF, IL-1β, IL-6, IL-12p40, IL-8, CCL2 and IL-10 in BDCA1+ DC supernatants | Circulating myeloid dendritic (CD1chigh) cells stimulated with LPS. | Not calculated | n = 52 ALS n = 36 healthy donors n = 25 other neurological controls. | Higher levels of IL-8 and CCL-2 upon LPS- stimulation in ALS dendritic cells | Rusconi et al., 2017 [54] |
 116 leukocyte populations and phenotypes from lymphocytes, monocytes, and granulocytes | Peripheral blood immunophenotyping by flow cytometry | 21.6 months | n = 80 ALS n = 50 controls | - 32 leukocyte phenotypes altered in ALS - elevated cell counts of granulocytes, NK cells and T cells in ALS - ALS patients were clustered into a profile distinct from controls primarily due to differences in multiple T cell phenotypes, CD3 CD56 T cells and HLA-DR on monocytes. | Gustafson et al., 2017 [55] |
 Transcriptomic analysis | RNA sequencing of blood monocytes | Not reported | n = 43 ALS n = 22 controls | ALS monocytes demonstrated a unique inflammation-related gene expression profile, the most prominent of which, including IL1B, IL8, FOSB, CXCL1, and CXCL2 | Zhao et al., 2017 [56] |
 IL1-b, IL-6, IL-8, IL-10, GM-CSF, and TNF and TGF-b1, −b2, and -b3 | By Luminex xMAP on supernatants from PBMCs or macrophages cultured overnight (non-stimulated, and SOD1-stimulated) | Not reported | 1 discordant twin pair | - In non-stimulated conditions the supernatants from the ALS PBMCs increased IL-6, TNF, and IL-1. | Lam et al., 2016 [57] |
 Immune and cytokine profiling | freshly collected, un-stimulated cells by flow cytometry, on peripheral monocytes and T lymphocytes. | 2.4 years | n = 24 ALS n = 25 controls | Th1-, Th17-, and IL-6-driven inflammation increased in ALS. | Saresella et al., 2013 [58] |
 90 inflammatory genes | qPCR analysis from isolated PBMCs | 25.4 months | n = 10 ALS n = 4 controls | - 50% of the ALS patients had ‘strong inflammation’ (upregulation of IL-1, IL-6, IL23a, PTGS2, MMP1, CCL20, CXCL3, CXCL5 and CXCR4; downregulation of CXCL9, CXCL10, and CXCL11), the other 50% had ‘weak inflammation’. - all ALS patients had an ‘ALS signature’ with 4-fold increase of MMP1, CCL7, CCL13 and CCL24. | Fiala et al., 2013 [59] No NCT reported |
 IL-1b, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IFN-γ, GM-CSF, TNF | PBMC cultures, non-stimulated and stimulated with SOD1 protein, supernatants analyzed by R&D Systems High Sensitivity Human Inflammation Multiplex-Kit | 26.9 months | n = 8 sALS n = 4 controls n = 1 unaffected twin of sALS patient | - 4 sALS patients had increased expression of TLR2 and CD14; ALOX5, PTGS2 and MMP1; IL1α, IL1β, IL6, IL36G, IL8 and TNF; CCL3, CCL20, CXCL2, CXCL3 and CXCL5. - 4 sALS patients had a decrease in the expression of PPARG, PPARA, RARG, HDAC4 and KAT2B; IL6R, IL6ST and ADAM17; TNFRSF11A; MGAT2 and MGAT3; PLCG1; CXCL3. - Difference identified between rapid ALS and slow ALS or controls. No diff between slow ALS and controls. | Mizwicki et al., 2012 [60] |
 monocyte and lymphocyte populations and activation | Surface expression, measured by flow cytometry from monocytes isolated from whole blood | 4–93 months (range) | n = 38 sALS n = 28 controls | - increased percentage of CD4+ cells in ALS - increased mean CD14-HLD-DR expression in ALS - increased percentage of CD14 and CD16+ cells in ALS - increased serum IgG in ALS - decreased serum IgM in ALS | Zhang et al., 2005 [61] |