Quantitative Transcriptome Analysis of Purified Equine Mast Cells Identifies a Dominant Mucosal Mast Cell Population with Possible Inflammatory Functions in Airways of Asthmatic Horses
Abstract
:1. Introduction
2. Results
2.1. Method to Isolate MCs from BALF of Asthmatic Horses
2.2. Transcriptome Analysis of Purified MCs and Total BALF Cells
2.3. Protease Profiles Reveal a Typical Mucosal MC Population in Horse BALF
2.4. Expression of FcεRI and Other MC Surface Markers
2.5. Low Expression of Toll-Like Receptors and Most Cytokine/Chemokine Receptors
2.6. Enzymes Related to Histamine and Arachidonic Acid Metabolites
2.7. Genes Related to Glycosaminoglycans
2.8. Expression of Transcription Factors
2.9. Markers Related to Antigen Presentation and other Immune Related Functions
3. Discussion
4. Materials and Methods
4.1. Horses and Collection of BALF
4.2. Mast Cell Isolation
4.3. Mast Cell Purity and Morphology
4.4. RNA Purification and Quantitative Transcriptome Analysis
4.5. RNA-Sequencing Data Analysis
4.6. Differential Gene Expression Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Genes | BALF MCs | Total BALF Cells | |
---|---|---|---|
Proteases | CMA1 | 0 | 0.4 |
CPA3 | 0 | 0 | |
TPSB2 | 1530 | 43 | |
TPSG1 | 16 | 0.3 | |
CTSG | 0 | 0 | |
GZMA | 68 | 166 | |
GZMK | 63 | 109 | |
GZMM | 2 | 2 | |
ELANE | 0 | 0 | |
AZU1 | 0 | 0 | |
NSP4 | 0 | 0 | |
MMP2 | 82 | 1.3 | |
MMP9 | 28 | 3 | |
CTSC | 275 | 100 | |
CPM | 18 | 44 | |
CPD | 11 | 5 | |
CPE | 0 | 0 | |
CPZ | 0 | 0 | |
CPA4 | 0 | 0 | |
CPA5 | 0 | 0 | |
CPA1 | 0 | 0 | |
CPB1 | 0 | 0 | |
CPA6 | 0 | 0 | |
CPB2 | 0 | 0 | |
Protease inhibitors | CST3 | 6198 | 565 |
SPINT2 | 244 | 16 | |
SERPINB9 | 237 | 18 | |
SERPINB6 | 230 | 27 |
Genes | BALF MCs | Total BALF Cells | |
---|---|---|---|
Immunoglobulin Receptors | FCER1A | 1764 | 43 |
FCER1G | 3387 | 4710 | |
MS4A2 | 147 | 1 | |
FCER2 | 5 | 0.6 | |
FCMR | 2 | 0.8 | |
Cytokine receptors | ST2 | 58 | 0.9 |
C-KIT | 164 | 1.4 | |
IL1R1 | 3 | 0.3 | |
IL1R2 | 5 | 0.6 | |
IL2RB | 60 | 20 | |
IL4R | 78 | 48 | |
IL9R | 6 | 0.1 | |
IL10RA | 47 | 28 | |
IL13RA1 | 54 | 18 | |
IL18R1 | 8 | 1 | |
IL21R | 53 | 4 | |
IL27RA | 11 | 77 | |
Chemokine receptors | CCR1 | 35 | 1.9 |
CCR2 | 21 | 14 | |
CCR3 | 5 | 6 | |
CCR5 | 13 | 5 | |
CCR7 | 650 | 26 | |
Toll-like receptors | TLR1 | 18 | 4 |
TLR2 | 16 | 47 | |
TLR3 | 13 | 11 | |
TLR4 | 0 | 0 | |
TLR5 | 1.2 | 0.2 | |
TLR6 | 8 | 5 | |
TRL7 | 17 | 16 | |
TLR8 | 13 | 32 | |
TLR9 | 2 | 0.4 | |
TLR10 | 13 | 0.2 | |
Other receptors | LY49B | 55 | 0.9 |
MILR1 | 13 | 4 | |
HRH4 | 24 | 0.4 |
Genes | BALF MCs | Total BALF Cells | |
---|---|---|---|
Heparin and Histamine synthesis | HDC | 166 | 2.5 |
HS6ST1 | 69 | 8 | |
HS3ST1 | 47 | 2 | |
HS6ST2 | 2 | 0 | |
NDST2 | 52 | 4 | |
NDST1 | 1 | 3.5 | |
SRGN | 5957 | 5935 | |
Chondroitin sulfate synthesis | CSGALNACT1 | 1.2 | 2 |
CSGALNACT2 | 21 | 7 | |
CHST3 | 0.1 | 0.6 | |
CHST7 | 45 | 5 | |
CHST11 | 1.5 | 10 | |
CHST13 | 32 | 2 | |
CHST15 | 1.5 | 0.4 | |
CHSY1 | 3 | 1.9 | |
CHSY3 | 0.03 | 2.5 | |
CHPF | 34 | 24 | |
Leukotriene synthesis | LTC4S | 2413 | 109 |
ALOX5 | 199 | 34 | |
ALOX5AP | 2174 | 1887 | |
Prostaglandin synthesis | HPGDS | 441 | 16 |
PTGS1 | 297 | 5 | |
PTGES3 | 216 | 211 | |
PTGDS | 18 | 0.9 | |
Transcription factors | GATA1 | 59 | 2 |
GATA2 | 93 | 2.2 | |
GATA3 | 20 | 5 |
Genes | BALF MCs | Total BALF Cells | |
---|---|---|---|
MHC-II and CD1 | DRA | 17,906 | 2344 |
DQA | 78 | 7 | |
DQB | 2988 | 375 | |
DRB | 1990 | 378 | |
HLA-DMA | 950 | 460 | |
HLA-DOB | 120 | 6 | |
CD74 | 14,764 | 3531 | |
CD1A3 | 426 | 44 | |
CD1A7 | 210 | 36 | |
CD1B1 | 157 | 5 | |
CD1B2 | 19 | 0.7 | |
CD1C | 44 | 0.9 | |
CD1E1 | 195 | 6 | |
CD1E2 | 537 | 18 | |
Complement components | C1QB | 1274 | 3896 |
C1QC | 1525 | 6168 | |
FCN1 | 48 | 1480 | |
CFP | 72 | 789 | |
Lysozyme | LYZ | 479 | 42 |
Genes | BALF MCs | Total BALF Cells | |
---|---|---|---|
Cytokine and Chemokines | TGFB1 | 460 | 434 |
IL1B | 7 | 3 | |
IL4 | 1 | 0.4 | |
IL4L1 | 83 | 17 | |
IL5 | 6 | 0.4 | |
IL6 | 9 | 3 | |
IL13 | 1 | 0 | |
IL15 | 6 | 12 | |
IL17 | 9 | 3 | |
IL18 | 68 | 160 | |
IL25 | 0 | 0 | |
IL33 | 0 | 0 | |
LIF | 46 | 1.3 | |
MIF | 631 | 862 | |
CCL5 | 163 | 483 | |
CCL8 | 242 | 242 | |
CCL22 | 59 | 2 | |
CCL24 | 268 | 1211 |
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Akula, S.; Riihimäki, M.; Waern, I.; Åbrink, M.; Raine, A.; Hellman, L.; Wernersson, S. Quantitative Transcriptome Analysis of Purified Equine Mast Cells Identifies a Dominant Mucosal Mast Cell Population with Possible Inflammatory Functions in Airways of Asthmatic Horses. Int. J. Mol. Sci. 2022, 23, 13976. https://doi.org/10.3390/ijms232213976
Akula S, Riihimäki M, Waern I, Åbrink M, Raine A, Hellman L, Wernersson S. Quantitative Transcriptome Analysis of Purified Equine Mast Cells Identifies a Dominant Mucosal Mast Cell Population with Possible Inflammatory Functions in Airways of Asthmatic Horses. International Journal of Molecular Sciences. 2022; 23(22):13976. https://doi.org/10.3390/ijms232213976
Chicago/Turabian StyleAkula, Srinivas, Miia Riihimäki, Ida Waern, Magnus Åbrink, Amanda Raine, Lars Hellman, and Sara Wernersson. 2022. "Quantitative Transcriptome Analysis of Purified Equine Mast Cells Identifies a Dominant Mucosal Mast Cell Population with Possible Inflammatory Functions in Airways of Asthmatic Horses" International Journal of Molecular Sciences 23, no. 22: 13976. https://doi.org/10.3390/ijms232213976