Expression of IL-8, IL-6 and IL-1β in Tears as a Main Characteristic of the Immune Response in Human Microbial Keratitis
Abstract
:1. Introduction
2. Results
2.1. Demographics, and Relevant Medical History of Patients with Microbial Keratitis
Characteristics of Patients | Patients with Fungal Keratitis (n = 14) | Patients with Gram-Positive Bacterial Keratitis (n = 8) | Patients with Gram-Negative Bacterial Keratitis (n = 6) | Healthy Volunteers (n = 48) | p Value |
---|---|---|---|---|---|
Males/Females | 10/4 | 3/5 | 3/3 | 24/24 | – |
Age (mean ± SD, years) | 49 ± 19 | 65 ± 19 | 51 ± 19 | 52 ± 19 | NS |
Microbiological diagnosis (number of patients) | Cephalosporium spp. (1) Curvularia spp. (1) Candida albicans (1) Trichoderma spp. (1) Aspergillus spp. (1) Fusarium spp. (9) | S. pneumonie (3) Streptococcus spp. (2) Micrococcus varians (1) S. pneumonie and S. epidermidis (2) | K. pneumonie (1) K. ozanae (1) Pseudomonas sp. (3) S. marcescens (1) | – | – |
Relevant medical history | Diabetes (4) Eye trauma (3) Diabetes and eye trauma (1) Alcoholism (1) Lens wear (1) None (4) | Diabetes (2) Previous herpes infection (2) Eye trauma (1) Congenital glaucoma (1) Rheumatoid arthritis (2) | Penetrating keratoplasty (1) Cosmetic lens/Lens wear (2) Diabetes (2) Drug user (1) | – | – |
2.2. Ocular Findings of Patients with Microbial Keratitis
Ocular Findings | Fungal Keratitis (n = 14) | Gram-Positive Bacterial Keratitis (n = 8) | Gram-Negative Bacterial Keratitis (n = 6) | p Value |
---|---|---|---|---|
Epithelial defect: Horizontal diameter (mm) | 5.3 ± 1.9 | 4.8 ± 1.9 | 5.6 ± 1.8 | NS between any groups of patients. |
Epithelial defect: Vertical diameter (mm) | 5.7 ± 2.2 | 5.5 ± 2.3 | 5 ± 2.4 | NS between any groups of patients. |
Stromal lesion: Horizontal infiltration diameter (mm) | 5.7 ± 2.4 | 4.9 ± 2.1 | 5 ± 2.4 | NS between any groups of patients. |
Stromal lesion: Vertical infiltration diameter (mm) | 6.0 ± 2.6 | 5.6 ± 2.3 | 4.3 ± 1.5 | NS between any groups of patients. |
Hypopyon (percentage) | 21 ± 22 | 37.5 ± 31 | 7.6 ± 8.6 | NS between any groups of patients. |
2.3. Frequency of CD3−CD56+ NK Cells Is Increased in Peripheral Blood Mononuclear Cells from Patients with Gram-Negative Bacterial Keratitis
2.4. IL-1β, IL-6, and IL-8 Are Increased in Tears Samples of Patients with Microbial Keratitis
PBMC Subpopulations | Fungal Keratitis (n = 14) | Gram-Positive Bacterial Keratitis (n = 8) | Gram-Negative Bacterial Keratitis (n = 6) | Healthy Volunteers (n = 48) | p Value |
---|---|---|---|---|---|
Percentage of CD3+ cells in PBMC | 57 ± 10 | 55 ± 8 | 63 ± 9 | 62 + 11 | NS between any groups |
Percentage CD4+ cells in PBMC | 34 ± 10 | 30 ± 3 | 31 ± 10 | 40 ± 13 | NS between any groups |
Percentage of CD8+ cells in PBMC | 25 ± 11 | 24 ± 10 | 26 ± 10 | 28 ± 12 | NS between any groups |
Percentage of CD19+ cells in PBMC | 12 ± 8 | 12 ± 9 | 10.1 ± 5 | 12 ± 7 | NS between any groups |
Percentage of CD3− CD56+ cells in PBMC | 28 ± 20 | 27 ± 13 | 39 ± 21 * | 25 ± 12 * | p < 0.05 |
CD4+IFN-γ+ cells (After PMA/ionomicyn stimulation) | 12 ± 10 | 16 ± 6 | 11 ± 6 | 10 ± 5 | NS between any groups |
CD4+IL-4+ (After PMA/ionomicyn stimulation) | 1 ± 1 | 3 ± 3 | 2 ± 2 | 4 ± 2 | NS between any groups |
CD8+IFN-γ+ (After PMA/ionomicyn stimulation) | 30 ± 27 | 16 ± 6 * | 31 ± 23 | 34 ± 20 * | p < 0.05 |
CD8+IL-4+ (After PMA/ionomicyn stimulation) | 1 ± 2 * | 1 ± 1 * | 2 ± 2 * | 10 ± 7 * | p < 0.05, when compared with healthy volunteers |
Tear Cytokine | Fungal Keratitis | Gram-Positive Bacterial Keratitis | Gram-Negative Bacterial Keratitis | Contralateral Non-Infected Eye | p Value (Compared with Contralateral Non-Infected Eye) |
---|---|---|---|---|---|
IL-1β pg/mL | 1135 ± 764 * | 64 ± 40 | 423 ± 240 * | 58 ± 89 * | <0.05 |
IL-6 pg/mL | 4172 ± 1873 * | 758 ± 1166 | 1596 ± 971 * | 25 ± 17 * | <0.05 |
IL-8 pg/mL | >2500 * | >2500 * | >2500 * | 420 ± 377 * | <0.05 |
IL-10 pg/mL | 43 ± 35 | 34 ± 23 | 107 ± 166 | 13 ± 3 | NS |
IL-12p70 pg/mL | 12 ± 6 | 12 ± 1 | 4.0 ± 1 | 5 ± 5 | NS |
TNF-α pg/mL | 28 ± 21 | 8 ± 4 | 8 ± 2 | 14 ± 7 | NS |
3. Discussion
4. Material and Methods
4.1. Patients
4.2. Clinical Evaluation
4.3. Monoclonal Antibodies and Reagents
4.4. Peripheral Blood Mononuclear Cells
4.5. Immunofluorescence Staining of Cell Surface Markers
4.6. Cell Cultures
4.7. Immunofluorescence Staining of Intracellular Cytokines
4.8. Flow Cytometric Analysis
4.9. Determination of Soluble Cytokines
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Santacruz, C.; Linares, M.; Garfias, Y.; Loustaunau, L.M.; Pavon, L.; Perez-Tapia, S.M.; Jimenez-Martinez, M.C. Expression of IL-8, IL-6 and IL-1β in Tears as a Main Characteristic of the Immune Response in Human Microbial Keratitis. Int. J. Mol. Sci. 2015, 16, 4850-4864. https://doi.org/10.3390/ijms16034850
Santacruz C, Linares M, Garfias Y, Loustaunau LM, Pavon L, Perez-Tapia SM, Jimenez-Martinez MC. Expression of IL-8, IL-6 and IL-1β in Tears as a Main Characteristic of the Immune Response in Human Microbial Keratitis. International Journal of Molecular Sciences. 2015; 16(3):4850-4864. https://doi.org/10.3390/ijms16034850
Chicago/Turabian StyleSantacruz, Concepcion, Marisela Linares, Yonathan Garfias, Luisa M. Loustaunau, Lenin Pavon, Sonia Mayra Perez-Tapia, and Maria C. Jimenez-Martinez. 2015. "Expression of IL-8, IL-6 and IL-1β in Tears as a Main Characteristic of the Immune Response in Human Microbial Keratitis" International Journal of Molecular Sciences 16, no. 3: 4850-4864. https://doi.org/10.3390/ijms16034850