A rapid method for the isolation of untouched naive CD8 T cells from peripheral blood mononuclear cells (46.9)
Bibliographic record
Abstract
Abstract Naïve CD8 T cells are, by definition, those that have been released by the thymus into circulation, but have not yet encountered their cognate antigens. Once they become activated they proliferate and generate effector T cells. We describe an easy and rapid method for the isolation of untouched naïve CD8 T cells from peripheral blood mononuclear cells (PBMC). Briefly, PBMCs are isolated by Ficoll-Paque PLUS density gradient sedimentation. Naïve CD8 T cells are then enriched using immunomagnetic, column-free, negative selection (EasySep™). Using EasySep™, unwanted cells are removed using antibody complexes recognizing non-naïve CD8 T cells and dextran-coated magnetic particles. Labeled cells are separated by placing the tube in an EasySep™ magnet. This procedure can be fully automated using RoboSep™. EasySep™ isolated naïve CD8 T cells are assessed by flow cytometry using a combination of antibodies specific for naïve, effector and memory cells. Naïve CD8 T cells are defined as CD8+CD45RA+CCR7+ and CD45RO-CD56-CD57-. Starting with PBMCs isolated from fresh blood and buffy coat, purities of 89±3% (n=8) are obtained (starting cell frequency 1.0-11.2%). EasySep™ isolated naïve CD8 T cells undergo robust proliferation as measured by flow cytometry of CFSE-labeled cells in culture with CD3/28 T cell expander beads. This simple method to isolate naïve CD8 T cells will be invaluable to researchers studying T cell differentiation, autoimmunity and infectious diseases.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.001 |
| Meta-epidemiology (narrow) | 0.002 | 0.001 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.002 | 0.001 |
| Science and technology studies | 0.001 | 0.000 |
| Scholarly communication | 0.001 | 0.001 |
| Open science | 0.002 | 0.001 |
| Research integrity | 0.001 | 0.002 |
| Insufficient payload (model declined to judge) | 0.014 | 0.028 |
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 source (direct Gemma or distilled Codex), 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".