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| Status |
Public on Dec 07, 2020 |
| Title |
Transcriptome analysis of mammary CD45+ CD4+ lymphocytes after immunization against E. coli P4 strain and homologous challenge |
| Organism |
Bos taurus |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Methods: During the last trimester of the gestation, three heifers were immunized using intramuscular injections of heat-killed E. coli P4 emulsified in Montanide ISA 61VG® adjuvant (SEPPIC). Two months later, they received a mammary ductal injection of 50 µg protein from concentrated E. coli supernate as a booster. Control cows were injected intramuscularly with adjuvant only at the same dates. Cows were then challenged on average at 140 days in milk in one healthy quarter by infusion of a E. coli P4 bacterial suspension (10e3 bacteria in total). Cows were then euthanized 16 hours after inoculation for mammary tissue collection. After collagenase and DNase treatment and Percoll-gradient separation, CD45+ CD4+ mononuclear cells were sorted by flow cytometry. RNA was extracted and sequenced using Illumina protocol.
Results : Mammary Th17-polarized CD4 T cells predominate in the mammary tissue of cows immunized through the local route, and are activated after challenge with E. coli.
Conclusions: Local immunization promotes the development of TH17 immunity that correlated with protection.
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| Overall design |
During the last trimester of the gestation, three heifers were immunized using intramuscular injections of heat-killed E. coli P4 emulsified in Montanide ISA 61VG® adjuvant (SEPPIC). Two months later, they received a mammary ductal injection of 50 µg protein from concentrated E. coli supernate as a booster. Control cows were injected intramuscularly with adjuvant only at the same dates. Cows were then challenged on average at 140 days in milk in one healthy quarter by infusion of a E. coli P4 bacterial suspension (10e3 bacteria in total). Cows were then euthanized 16 hours after inoculation for mammary tissue collection. After collagenase and DNase treatment and Percoll-gradient separation, CD45+ CD4+ mononuclear cells were sorted by flow cytometry. RNA was extracted and sequenced using Illumina protocol.
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| Contributor(s) |
FOUCRAS G, CEBRON N, MAMAN S, RAINARD P |
| Citation(s) |
33298970 |
|
https://doi.org/10.1038/s41541-020-00258-4
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| |
| Submission date |
Oct 09, 2020 |
| Last update date |
Dec 15, 2020 |
| Contact name |
gilles FOUCRAS |
| E-mail(s) |
[email protected]
|
| Organization name |
ENVT INRA
|
| Street address |
23 Chemin des Capelles
|
| City |
TOULOUSE |
| State/province |
State/Province/Region |
| ZIP/Postal code |
31300 |
| Country |
France |
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| Platforms (1) |
| GPL21659 |
Illumina HiSeq 3000 (Bos taurus) |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA668294 |
| SRA |
SRP286865 |
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