Introduction
Materials and methods
Patients and bronchoscopy
Analyses of BAL fluid
Lymphocyte subset | Antibody ligand (fluorochrome) |
---|---|
T cells (CD3+) | CD3 (PerCP) |
T helper cells (CD3+CD4+) | CD3 (PerCP), CD4 (FITC) |
Cytotoxic T cells (CD3+CD8+) | CD3 (PerCP), CD8 (PE) |
Natural killer cells (CD3−CD16+CD56+) | CD3 (FITC), CD16 (PE), CD56 (PE) |
Regulatory T cells (CD3+CD4+CD25brightCD127low/-) | CD3 (APC), CD4 (FITC), CD25 (PE-Cy5), CD127 (PE) |
Activated T helper cells (CD3+CD4+CD25+) | CD3 (APC), CD4 (FITC), CD25 (PE) |
Activated T helper cells (CD3+CD4+CD69+) | CD3 (APC), CD4 (FITC), CD69 (PE) |
Activated T helper cells (CD3+CD4+HLA-DR+) | CD3 (APC), CD4 (FITC), HLA-DR (PE) |
Activated cytotoxic T cells (CD3+CD8+CD25+) | CD3 (APC), CD8 (PerCP), CD25 (PE) |
Activated cytotoxic T cells (CD3+CD8+CD69+) | CD3 (APC), CD8 (PerCP), CD69 (PE) |
Activated cytotoxic T cells (CD3+CD8+HLA-DR+) | CD3 (APC), CD8 (PerCP), HLA-DR (PE) |
Activated cytotoxic T cells (CD3+CD8+NKG2D+) | CD3 (PerCP), CD8 (FITC), NKG2D (PE) |
Indicators of disease severity
Statistical analysis
Results
Clinical and laboratory findings
Days of hospitalization | 5 (2–9) |
Clinical findings | |
Hypotension (≤90 mmHg) | 6 (35 %) |
Respiratory symptoms | 10 (59 %) |
Dyspnea | 8 (47 %) |
Dry cough | 5 (29 %) |
Oxygen treated | 5 (29 %) |
Laboratory results | |
Leukocyte count (3.5–8.8 × 109/L), max | 9.0 (5.3–27.0) |
C-reactive protein (<3 mg/L), max | 78 (35–249) |
Platelet count (145–387 × 109/L), min | 63 (18–305) |
Creatinine (<105 μmol/L), max | 173 (59–1072) |
Lactate dehydrogenase (<3.4 μkat/L), max | 4.8 (3.8–12.3) |
Albumin (36–45 g/L), min | 28 (14–34) |
NT-proBNP (<150 ng/L), max | 1768 (121–8878) |
Troponin T (<15 ng/L), max | 8 (0–22) |
Oxygen treatment (n = 5) | No oxygen treatment (n = 12) |
p-Value | |
---|---|---|---|
Laboratory results | |||
Leukocyte count (109/L), max | 15.1 (9.1–15.8) | 8.6 (7.0–9.5) | 0.058 |
C-reactive protein (mg/L), max | 187 (65–203) | 73 (37–128) | 0.11 |
Platelet count (109/L), min | 63 (42–85) | 63 (42–90) | 1.00 |
Creatinine (μmol/L), max | 276 (186–327) | 130 (83–219) | 0.058 |
Lactate dehydrogenase (μkat/L), max | 4.9 (4.6–6.2) | 4.5 (4.2–5.1) | 0.21 |
Albumin (g/L), min | 22 (17–27) | 29 (24–31) | 0.034 |
NT-proBNP (ng/L), max | 2342 (1419–6862) | 728 (523–4393) | 0.17 |
Troponin T (ng/L), max | 10 (7–19) | 8 (0–11) | 0.22 |
Differential cell counts | |||
Macrophages | 22.0 (17.1–44.3) | 15.0 (11.4–30.2) | 0.21 |
Eosinophils | 0.1 (0–0.2) | 0 (0–0.8) | 0.21 |
Neutrophils | 0.6 (0.5–0.7) | 0.4 (0.2–0.6) | 0.14 |
Lymphocytes | 9.9 (6.2–15.7) | 3.2 (2.0–7.7) | 0.015 |
Flow cytometry | |||
T cells | 9.5 (5.6–14.9) | 2.6 (1.5–7.5) | 0.020 |
T helper cells | 1.9 (1.7–5.2) | 0.6 (0.3–2.7) | 0.079 |
Cytotoxic T cells | 7.3 (3.5–9.3) | 1.4 (1.1–4.8) | 0.036 |
Natural killer cells | 0.5 (0.4–0.6) | 0.2 (0.1–0.5) | 0.10 |
Regulatory T cells | 0.6 (0.4–1.5) | 0.4 (0.2–0.7) | 0.19 |
T helper cells, CD25+
| 0.1 (0.1–0.2) | 0 (0–0.1) | 0.047 |
T helper cells, CD69+
| 0 (0–0) | 0 (0–0.3) | 0.82 |
T helper cells, HLA-DR+
| 0.5 (0.3–1.3) | 0.1 (0.1–0.4) | 0.066 |
Cytotoxic T cells, CD25+
| 0.1 (0–0.5) | 0.1 (0–0.1) | 0.40 |
Cytotoxic T cells, CD69+
| 0 (0–0.2) | 0.1 (0–0.2) | 0.96 |
Cytotoxic T cells, HLA-DR+
| 3.6 (1.5–4.1) | 0.9 (0.4–2.9) | 0.11 |
Cytotoxic T cells, NKG2D+
| 2.6 (1.0–4.3) | 0.6 (0–1.0) | 0.020 |
Granzymes and cytokines | |||
Granzyme A | 326 (104–643) | 255 (116–509) | 0.83 |
Granzyme B | 33 (6–84) | 5 (0–23) | 0.070 |
Interleukin 6 | 0.8 (0.7–1.8) | 0.9 (0.6–1.5) | 0.83 |
Tumor necrosis factor-α | 0 (0–0) | 0 (0–0.6) | 0.24 |
Viral load | 1.4 × 101 (0.2 × 101 to 6.2 × 105) | 3.0 × 103 (1.2 × 101 to 1.5 × 104) | 0.40 |
Cytotoxic responses in the lungs
Patients (n = 17) | Controls (n = 16) |
p-Value | |
---|---|---|---|
Return volume | 95.0 (81.0–119.0) | 108.0 (67.5–123.0) | 0.86 |
Differential cell counts | |||
Macrophages | 16.8 (11.7–33.5) | 20.5 (13.4–32.1) | 0.67 |
Neutrophils | 0.5 (0.2–0.6) | 0.2 (0.0–0.4) | 0.088 |
Eosinophils | 0.00 (0.00–0.11) | 0.05 (0.00–0.28) | 0.40 |
Lymphocytes | 4.2 (2.1–9.3) | 1.8 (1.2–2.5) | 0.001 |
Flow cytometry | |||
T cells | 4.3 (1.9–9.1) | 1.5 (1.1–1.8) | 0.002 |
T helper cells | 1.3 (0.4–2.5) | 1.0 (0.6–1.5) | 0.71 |
Cytotoxic T cells | 2.6 (1.1–6.4) | 0.3 (0.1–0.6) | <0.001 |
Natural killer cells | 0.08 (0.05–0.11) | 0.04 (0.02–0.07) | 0.034 |
Regulatory T cells | 0.42 (0.22–0.80) | 1.38 (0.77–3.14) | 0.004 |
T helper cells, CD69+
| 0.02 (0.00–0.04) | 0.01 (0.00–0.07) | 0.62 |
T helper cells, HLA-DR+
| 0.23 (0.06–0.59) | 0.20 (0.12–0.32) | 0.46 |
T helper cells, CD25+
| 0.06 (0.02–0.14) | 0.06 (0.02–0.14) | 0.80 |
Cytotoxic T cells, CD25+
| 0.06 (0.02–0.19) | 0.00 (0.00–0.01) | <0.001 |
Cytotoxic T cells, CD69+
| 0.05 (0.01–0.16) | 0.01 (0.00–0.03) | 0.046 |
Cytotoxic T cells, HLA-DR+
| 1.14 (0.46–3.12) | 0.03 (0.01–0.06) | <0.001 |
Cytotoxic T cells, NKG2D+
| 0.80 (0.06–2.11) | 0.04 (0.01–0.05) | 0.001 |
Granzymes and cytokinesa
| |||
Granzyme A | 287.5 (126.3–484.5) | 19.7 (13.7–59.4) | <0.001 |
Granzyme B | 6.3 (0–30.4) | 0 (0–0) | <0.001 |
Interleukin 6 | 0.8 (0.7–1.5) | 1.6 (0.9–2.1) | 0.08 |
Tumor necrosis factor-α | 0 (0–0) | 0 (0–0) | 0.61 |