Efficient isolation of particle-free human ILC2s from peripheral blood mononuclear cells
Bibliographic record
Abstract
Abstract Group 2 innate lymphoid cells (ILC2s) have important roles in type-2 immune responses and are implicated in allergies, asthma, helminth infections and other metabolic diseases. To better understand their role in immunity, highly purified ILC2s are required. ILC2s are phenotypically defined as CD45+ lineage-negative CRTH2+ CD127+ CD161+, and comprise approximately 0.05% of peripheral blood mononuclear cells (PBMCs). Fluorescence activated cell sorting is the most common method to isolate ILC2s but it is both time-consuming and expensive because the cells are so rare. To address this issue, we have developed an immunomagnetic isolation method to obtain highly pure, particle-free ILC2s from human PBMCs in 2.5 hours. First, CRTH2 positive cells are labelled with a CRTH2-PE monoclonal antibody. These cells are then positively selected using an anti-PE antibody complex, EasySep™ Releasable RapidSpheres™ and an EasySep™ magnet. After separation, the magnetic particles are released from the positively selected cells and a second cocktail of antibody complexes and EasySep™ Dextran RapidSpheres™ is added to label non-ILC2s within the CRTH2 positive population. After a second separation, the released particles and unwanted cells are retained in the tube within the magnet, whereas the ILC2s are simply poured off and ready for use. With this method, the purity of ILC2s isolated from PBMCs was 84–95% (median 91%, n=18) with a recovery of 0.4–14.3×103 ILC2s per 108 starting PBMCs (median 2.46×103, n=18). The isolated ILC2s are functional, producing IL-13 upon in vitro stimulation with IL-33 and IL-2. Overall, EasySep™ Human ILC2 Isolation Kit allows researchers to easily and efficiently isolate highly pure and functional ILC2s.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.001 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.001 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.001 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".