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Thymic function in MHC class II–deficient patients

Published byFarida Veronika Indradjaja Modified over 5 years ago

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1 Thymic function in MHC class II–deficient patients
Atar Lev, MSc, Amos J. Simon, BSc, Arnon Broides, MD, Jacob Levi, MD, Ben Zion Garty, MD, Ester Rosenthal, MSc, Ninette Amariglio, PhD, Gideon Rechavi, MD, PhD, Raz Somech, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 131, Issue 3, Pages (March 2013) DOI: /j.jaci Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 TCR repertoire: FACS analyses on CD3+CD4/CD8+ cells. Relative expression of 24 different TCR-Vβ families in CD3+CD4+ cells (A) and CD3+CD8+ cells (B; solid bars) obtained from 4 MHC-II–deficient patients' PBMCs compared with the relative expression of normal CD3+CD4+ or CD3+CD8+ control cells (open bars). Normal control values were obtained from the IOTest Beta Mark-Quick Reference Card (Beckman Coulter, Fullerton, Calif). P, Patient. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 1 TCR repertoire: FACS analyses on CD3+CD4/CD8+ cells. Relative expression of 24 different TCR-Vβ families in CD3+CD4+ cells (A) and CD3+CD8+ cells (B; solid bars) obtained from 4 MHC-II–deficient patients' PBMCs compared with the relative expression of normal CD3+CD4+ or CD3+CD8+ control cells (open bars). Normal control values were obtained from the IOTest Beta Mark-Quick Reference Card (Beckman Coulter, Fullerton, Calif). P, Patient. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 2 TCR repertoire: FACS analyses on CD3+CD4/CD8+CD45RO/CD45RA+ cells. Relative expression of 24 different TCR-Vβ families in CD3+CD4/CD8+CD45RA+ (solid bars) and CD3+CD4/CD8+CD45RO+ (shaded bars) cells obtained from 2 MHC-II–deficient patients' PBMCs compared with the relative expression of normal CD3+CD4+ or CD3+CD8+ control cells (open bars). Normal control values were obtained from the IOTest Beta Mark-Quick Reference Card (Beckman Coulter). A, CD4 cells from patient 5. B, CD8 cells from patient 5. C, CD4 cells from patient 8. D, CD8 cells from patient 8. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 3 Rearrangement analysis of TCR-Vγ in MHC-II–deficient patients' PBMCs. Four different Vγ rearranged TCR genes (Vγ9.2, Vγ11, VγF1, and Vγ10.2) were PCR amplified in 8 MHC-II–deficient patients and age-matched healthy control subjects, followed by Gene Scan analysis. Four different patterns are shown: p, polyclonal; s, skewed polyclonal; o, oligoclonal; u, undetected. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 4 TREC copy numbers in dried blood spots. TRECs were determined on Guthrie cards with dried blood spots obtained from 15 anonymized healthy infants (open diamonds), 7 patients with SCID (open circles), and 2 MHC-II–deficient patients (solid diamonds). The dashed line represents cutoff values for TREC levels per 3-mm punched disk. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 5 Analysis of the ordered TCR rearrangement events in MHC-II–deficient patients' PBMCs. Five different TCR rearrangement events (TCRD-Dδ2-Dδ3, TCRD-Dδ2-Jδ1, TCRD-Vδ1-Jδ1, TCRA-REC, and TREC) were analyzed by using qRT-PCR in PBMCs obtained from 5 MHC-II–deficient patients and 7 age-matched healthy control subjects. The control is presented as the average ± SD of the 7 samples. For each patient, the TCR rearrangements are presented as DNA copies in log scale. The obtained Ct in each detected point was compared with the Ct of the TREC copies in the different standard serial dilutions, as described in the text (A). For each TCR rearrangement, the averages of the results obtained either for the 5 patients or for the 7 control subjects were calculated and compared. SDs of the calculated averages are included (B). Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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