Unbiased Analysis of Temporal Changes in Immune Serum Markers in Acute COVID-19 Infection With Emphasis on Organ Failure, Anti-Viral Treatment, and Demographic Characteristics

Abstract

Identification of novel immune biomarkers to gauge the underlying pathology and severity of COVID-19 has been difficult due to the lack of longitudinal studies. Here, we analyzed serum collected upon COVID-19 admission (t1), 48 hours (t2), and seven days later (t3) using Olink proteomics and correlated to clinical, demographics, and therapeutic data. Older age positively correlated with decorin, pleiotrophin, and TNFRS21 but inversely correlated with chemokine (both C-C and C-X-C type) ligands, monocyte attractant proteins (MCP) and TNFRS14. The burden of pre-existing conditions was positively correlated with MCP-4, CAIX, TWEAK, TNFRS12A, and PD-L2 levels. Individuals with COVID-19 demonstrated increased expression of several chemokines, most notably from the C-C and C-X-C family, as well as MCP-1 and MCP-3 early in the course of the disease. Similarly, deceased individuals had elevated MCP-1 and MCP-3 as well as Gal-9 serum levels. LAMP3, GZMB, and LAG3 at admission correlated with mortality. Only CX3CL13 and MCP-4 correlated positively with APACHE score and length of stay, while decorin, MUC-16 and TNFRSF21 with being admitted to the ICU. We also identified several organ-failure-specific immunological markers, including those for respiratory (IL-18, IL-15, Gal-9) or kidney failure (CD28, VEGF). Treatment with hydroxychloroquine, remdesivir, convalescent plasma, and steroids had a very limited effect on the serum variation of biomarkers. Our study identified several potential targets related to COVID-19 heterogeneity (MCP-1, MCP-3, MCP-4, TNFR superfamily members, and programmed death-ligand), suggesting a potential role of these molecules in the pathology of COVID-19.

Keywords: CCL23; COVID-19; MCP; convalescent plasma; hydroxychloroquine; programmed death; remdesavir; steroids.

Figure 1

Visualization of temporal changes in the variation of the immunological biomarkers when age (cutoff of 60 years) (A), gender (B) and race (C) were compared across the study subjects. Summary of statistically significant differences demonstrated as higher (red) or lower (blue) than an appropriate comparison group at standard [(p ≤ 0.05); pale shade] and more conservative (darker shade) The mean and SD of the compared group are listed in Supplemental Material 2 . Denotation of abbreviations and corresponding UniProt number are listed in Supplemental Material 2 .

Figure 2

Regression analysis between pre-COVID-19 health conditions, calculated as the CCI and serum level of MCP-4 (A) and PD-L2 (B) when measured at admission (t1; red), 48 hours after admission (t2; yellow) and seven days after admission (t3; green).

Figure 3

Summary of statistically significant differences demonstrated as higher (red) or lower (blue) than an appropriate comparison group at standard [(*p < 0.05); pale shade] and more conservative (darker shade) The mean and SD of the compared group are listed in Supplemental Material 2 . Denotation of abbreviations and corresponding UniProt number are listed in Supplemental Material 2 .

Figure 4

Also, patients who were admitted to the ICU had distinguish inflammatory profile at the admission (A) with DCN, NCR1, IL-15 and TNFRS21 being good predictor of ICU admission (B).

Figure 5

Intubation was signified for several markers (A) which also discriminate patient at the admission for those requiring ventilatory support (B). The profile of patient requiring ECMO was significantly different considering large number of inflammatory marker being depressed as compared to patient not requiring ECMO (C).

Figure 6

Several markers correlated strongly with numerous measures of COVID-19 severity (A). CXCL-13 (B, C) and CCL23 (D, E) demonstrated strong correlations with admission APACHE_1hr (B, D) and the length of stay in the hospital (C, D). Summary of statistically significant differences demonstrated as higher (red) or lower (blue) correlation. LOS, length of stay; ICU, intensive care unit; Vent, mechanical ventilation; APACHE, acute physiology and chronic health evaluation; SOFA, sequential organ failure assessment; MODS, multiple organ dysfunction syndrome.

Figure 7

Time-related temporal markers of the immune system for cardiovascular failure (CVf; A), central nervous system failure (CNSf; B), and liver failure (Lf; C). Summary of statistically significant differences demonstrated as higher (red) or lower (blue) than an appropriate comparison group at standard [(p ≤ 0.05); pale shade] and more conservative (darker shade) The mean and SD of the compared group are listed in Supplemental Material 2 . Denotation of abbreviations and corresponding UniProt number are listed in Supplemental Material 2 .

Figure 8

Time-related temporal markers of the immune system for cardiovascular failure (Rf) (A) demonstrated IL-18, GAL-9, CXCL13 and TNFRS2 being predictors of patients requiring ventilatory support (B). Summary of statistically significant differences demonstrated as higher (red) or lower (blue) than an appropriate comparison group at standard [(p ≤ 0.05); pale shade] and more conservative (darker shade) The mean and SD of the compared group are listed in Supplemental Material 2 . Denotation of abbreviations and corresponding UniProt number are listed in Supplemental Material 2 .

Figure 9

Time-related temporal markers of the immune system for cardiovascular failure (AKIf) (A) demonstrated VEGFA and CD28 being predictors of patients experiencing acute kidney failure during hospitalization (B).Summary of statistically significant differences demonstrated as higher (red) or lower (blue) than an appropriate comparison group at standard [(p ≤ 0.05); pale shade] and more conservative (darker shade) The mean and SD of the compared group are listed in Supplemental Material 2 . Denotation of abbreviations and corresponding UniProt number are listed in Supplemental Material 2 .

Figure 10

The effect of hydroxychloroquine (A), remdesivir (B), steroids (C), and convalescent plasma (D) on selected biomarkers (complete list in Supplemental Figure 2 ). *denotes the difference p ≤ 0.05, **denotes the difference p ≤ 0.01.