Development and characterization of anti-Gal B cell receptor transgenic Gal???/??? mice
Bibliographic record
Abstract
BACKGROUND: The successful clinical application of pig-to-primate xenotransplantation is currently limited by the development of an acute vascular rejection, which is thought to involve an induced humoral immune response to the galactose alpha1,3 galactose (alpha-Gal) antigen. Successful xenotransplantation may require the development of novel methods for removal or neutralization of anti-Gal antibodies and anti-Gal-producing B cells. The large diversity of the B-cell repertoire makes it difficult, however, to isolate and study anti-Gal B-cell development. METHODS: We have established a transgenic mouse model for investigating anti-Gal B cells by introducing a transgene encoding both heavy and light chains for an anti-Gal IgM antibody into an alpha-galactosyltransferase-deficient (Gal-/-) background. We have characterized the frequency, phenotype, and function of transgenic anti-Gal B cells by multiparameter flow cytometric analysis and ELISA. RESULTS: ELISA analysis of serum from animals with the transgene in an alpha-galactosyltransferase-deficient background (Tg Gal-/-), from transgenic animals with a heterozygous alpha-galactosyltransferase background (Tg Gal-/+), and from nontransgenic alpha-galactosyltransferase-deficient littermates (Gal-/-) demonstrated elevated expression of anti-Gal antibodies in Tg Gal-/- mice compared with nontransgenic Gal-/- animals and a lack of transgene expression in the Tg Gal-/+ mice. Anti-Gal antibody expression in Tg Gal-/- mice could be increased by immunization with an ovalbumin-Gal glycoconjugate in vivo and through stimulation with lipopolysaccharide in vitro. Multiparameter flow cytometric analysis indicates that 50% to 80% of splenic and peritoneal B cells expressed the transgene and excluded endogenous immunoglobulin gene rearrangements. The majority of these B cells expressed anti-Gal receptors on the surface, as identified by staining with a fluorescein isothiocyanate-bovine serum albumin-Gal glycoconjugate. FACS analysis of the Tg Gal-/- B cells identified them as a population of CD21highCD23lowIgMhigh marginal zone B cells in the spleen and CD5-CD23low B1 cells in the peritoneal cavity. CONCLUSIONS: These observations suggest that this model can be used to study the regulation of anti-Gal B cells and can establish a reliable source of functional anti-Gal B cells, which could be used to test the effectiveness of alpha-Gal-specific immunosuppressive reagents.
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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 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.001 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.002 | 0.001 |
| Science and technology studies | 0.001 | 0.001 |
| Scholarly communication | 0.001 | 0.001 |
| Open science | 0.002 | 0.001 |
| Research integrity | 0.002 | 0.003 |
| Insufficient payload (model declined to judge) | 0.004 | 0.004 |
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".