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Blood, Vol. 108, Issue 6, 1941-1948, September 15, 2006
Aire-deficient mice develop hematopoetic irregularities and marginal zone B-cell lymphoma
Blood Hässler et al.
108: 1941
Supplemental materials for: Hassler et al
Files in this Data Supplement:
- Supplemental materials and methods (PDF, 23.5 KB) -
Included here are the measurement of immunoglobulin subclasses, in vitro transcription translation assay of APS I autoantigens, a hematological evaluation, a culture of Candida albicans, clinical chemistry and endocrinological measurements, and a description of B-cell transfer.
- Supplemental data (PDF, 27 KB) -
Included here are sera from Aire-/- mice that do not recognize APS I autoantigens and a hematological evaluation of Aire-/- mice. Among the findings, aged Aire-/- mice have a normal endocrine function in APS I target glands, and B cells from Aire-/- spleens transfer antibody production to Aire sufficient recipients.
- Table S1. Red blood cell and platelet parameters in blood (PDF, 11.7 KB)
- Figure S1. Immunoglobulin subclass levels of Aire deficient mice (PDF, 18.5 KB) -
Ig subclasses were measured with ELISA in sera from 15 months old Aire WT and KO mice. Each symbol corresponds to a different animal, and median is indicated by horizontal lines. P-values calculated with Mann-Whitney test. N=6 WT, 6 KO.
- Figure S2. Absence of autoantibodies against known APS I autoantigens in Aire deficient mice (PDF, 21.3 KB) -
Radioactive autoantigens synthesized by in vitro transcription and translation were immunoprecipitated with sera from either Aire WT or KO mice and radioactivity was measured by scintillation counting. Serum from an APS I patient was used as positive control. (A) 21-hydroxylase (21-OH), 17-hydroxylase (17-OH), side chain cleavage enzyme (SCC), aromatic L-amino acid decarboxylase (AADC) in 15 months old mice. N=10 WT, 11 KO. (B) Insulinoma associated tyrosine phosphatase like protein (IA2), cytochrome P450 1A2 (CYP1A2) and tryptophan hydroxylase (TH) in 6 months old mice. N=5 WT, 6 KO. Each symbol corresponds to a different animal, and median is indicated by horizontal lines. P-values calculated with Mann-Whitney test.
- Figure S3. Normal bone marrow hematopoiesis in aged Aire deficient mice (PDF, 14.4 KB) -
Differential count of myeloid and erythrocytic lineage cells represented as myeloid/erythroid (M/E) ratio; number of megakaryocytes in bone marrow cytospins. N=8 WT, 8 KO. Each symbol corresponds to a different animal, p-value calculated with t-test for megakaryocytes and Mann-Whitney test for M/E ratio. Mean values (t-test) or median (Mann-Whitney test) are indicated by horizontal lines.
- Figure S4. Endocrinological evaluation of Aire deficient mice (PDF, 17.3 KB) -
(A) Ca2+ and parathyroid hormone (PTH) were measured in sera by a colorimetric assay and ELISA respectively. N=11 WT, 9 KO (Ca2+); N=9 WT, 10 KO (PTH). (B) Corticosterone was measured in serum by ELISA. N=11 WT, 11 KO. (C) Analysis of alanine aminotransferase (ALAT) in mouse sera. N=4 WT, 9 KO. Each symbol corresponds to a different animal, p-values calculated with t-test for (A), with Mann-Whitney test for (B) and (C). Mean values (A) or median (B,C) are indicated by horizontal lines.
- Figure S5. B cells from Aire deficient spleens transfer antibody production to Aire sufficient recipients (PDF, 23 KB) -
Indirect immunofluorescent staining of adrenocortical cells with serum samples from Aire WT mice taken 6 days after transfer of splenic B cells from either WT or KO donors.
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