The modulatory action of C-Vx substance on the immune system in COVID-19

Figures & data

Table 1. Demographic characteristics of COVID-19 patients and treatments they received.

	Mild	Moderate	Severe
N	10	11	10
Gender	5 males	5 males	5 males
	5 females	6 females	5 females
Age Median (Min-Max)	45 (21-79)	54.8 (33-78)	63.2 (50-73)
Treatment received for COVID-19	Hydroxychloroquine, Azithromycin	Hydroxychloroquine, Azithromycin, Favipravir	Hydroxychloroquine, Azithromycin, Favipravir, Tociluzumab
The clinical symptoms of COVID-19 patients	Who do not require hospitalization with Fever, sore throat, dry cough, malaise, and body aches or nausea, vomiting, abdominal pain, loose stools	Who have moderate/severe pneumonia or mild pneumonia without hypoxemia with high ferritin levels high – C reactive protein (hsCRP) levels, lymphopenia, hypotension dyspnea	Who are hospitalized pneumonia with hypoxemia (SpO2< 92%) with coagulation defects, encephalopathy, heart failure, and acute kidney injury

Table 2. Biochemical parameters of patients.

	Mild Median (min-max)	Moderate Median (min-max)	Severe Median (min-max)	Reverences Values
White Blood Cells (10³/µL)	8.03 (3.9-13.2)	6.6 (3.4-11)	10.4 (5-16.6)	4-10
Lymphocytes (10³/µL)	1.55 (0.6-3.1)	1.26 (0.5-2.4)	1.35 (0.2-5.9)	1.2-3.6
Neutrophils (10³/µL)	5.88 (2.1-11.8)	4.96 (1.7-10.1)	7.93 (0.6-15.4)	1.3-7
Platelets (10³/µL)	276.3 (177-528)	238.1 (124-364)	261.9 (123-476)	160-390
CRP (mg/dL)	40.1 (0-164)	58.9 (3-150)	153.7 (13-528)	0-5
D-Dimer (µg/L)	968.9 (190-3700)	922.7 (220-1640)	4370 (380-18450)	0.0-550
Ferritin (ng/mL)	192.9 (22-842)	572.3 (24-1404)	6068.6 (232-46852)	13-400
Procalcitonin (ng/mL)	0.08 (0.01-0.19)	0.1 (0.01-0.35)	10.97 (0.07-48.89)	0-0.5

Figure 1. The evaluation of the effect of C-Vx on T and NK cell ratios and NK cell cytotoxicity. (A) CD3⁺ total T, (B) CD3⁻CD16⁺CD56⁺ NK, (C) CD4⁺, (D) CD8⁺ T cell frequencies, and (E) CD107a expression of NK cells in patient groups and healthy donors with and without C-Vx. (*p < 0.05, **p < 0.001, ***p < 0.0001) (US: unstimulated, HD: healthy donors).

Figure 2. The alterations in intracellular cytokine levels of T cell subsets in response to C-Vx. IFN-γ, IL-17, TNF-α, IL-4, and IL-10 levels of (A) CD4⁺ and (B) CD8⁺ T cells after culture of PBMCs stimulated or not with C-Vx. (*p < 0.05, **p < 0.001, ***p < 0.0001) (US: unstimulated, HD: healthy donors).

Figure 3. Determination of lymphocyte proliferation in response to C-Vx. The proliferation capacities of (A) total lymphocytes, (B) CD3⁺ T cells, (C) CD3⁻CD16⁺CD56⁺ NK cells, (D) CD4⁺, and (E) CD8⁺ T cell subsets after cell culture with/without PHA stimulation with or without C-Vx. (*p < 0.05, **p < 0.001, ***p < 0.0001) (US: unstimulated, HD: healthy donors).

Figure 4. Bradykinin, IRF3, and IFN-α levels in response to C-Vx. The effects of C-Vx addition on (A) IRF3, (B) Bradykinin, and (C) IFN-α levels in plasma and culture supernatants were measured by ELISA method (*p < 0.05, **p < 0.001, ***p < 0.0001) (US: unstimulated, HD: healthy donors).

Figure 5. The alterations in cytokine levels after C-Vx addition. IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, IFN-γ, TNF-α, and GM-CSF levels were measured by multiplex assay in supernatants of cultured PBMCs of patients and healthy donors with/without C-Vx. (*p < 0.05, **p < 0.001, ***p < 0.0001) (US: unstimulated, HD: healthy donors).

Figure 6. The network analysis of immune parameters in COVID-19 patients with/without C-Vx. Blue and red lines indicate positive and negative correlations. The significance of correlations increases in proportion with increased darkness of line colours. (US: unstimulated).