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Tight junction proteins expression and modulation in immune cells and multiple sclerosis.

Mandel I, Paperna T, Glass-Marmor L, Volkowich A, Badarny S, Schwartz I, Vardi P, Koren I, Miller A - J. Cell. Mol. Med. (2012)

Bottom Line: The tight junction proteins (TJPs) are major determinants of endothelial cells comprising physiological vascular barriers such as the blood-brain barrier, but little is known about their expression and role in immune cells.Interestingly, claudin 1 protein levels were elevated also in PBLs of patients with type 1 diabetes (T1D).Furthermore, a correlation between CLDN5 pre-treatment levels and clinical response phenotype to interferon-β therapy was detected.

View Article: PubMed Central - PubMed

Affiliation: Rappaport Faculty of Medicine and Research Institute, Technion-Israel Institute of Technology, Haifa, Israel.

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Expression levels of claudins 1 and 5 in leukocytes from MS and T1D. (A) Claudin 1 protein levels in PBLs of healthy controls (n = 13), OND controls (n = 14), T1D controls (n = 14), MS patients in remission (n = 13) and MS patients in relapse (n = 10). (B) The expression levels of CLDN5 RNA (2ΔCT) relative to reference genes were determined by real- time RT-PCR of PBL RNA derived from MS patients in relapse (n = 33), MS patients in remission (n = 39), and healthy controls (n = 18). (C) Claudin 5 protein levels in the PBLs of healthy controls (n = 13), OND controls (n = 14), T1D controls (n = 14), MS patients in remission (n = 11), and MS patients in relapse (n = 17). Box plot and data points display the relative protein levels determined by Western blot analysis. The relative intensity of claudin 1 or claudin 5 proteins compared to β-actin signals was determined by densitometry. Medians are depicted by horizontal bars.
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fig03: Expression levels of claudins 1 and 5 in leukocytes from MS and T1D. (A) Claudin 1 protein levels in PBLs of healthy controls (n = 13), OND controls (n = 14), T1D controls (n = 14), MS patients in remission (n = 13) and MS patients in relapse (n = 10). (B) The expression levels of CLDN5 RNA (2ΔCT) relative to reference genes were determined by real- time RT-PCR of PBL RNA derived from MS patients in relapse (n = 33), MS patients in remission (n = 39), and healthy controls (n = 18). (C) Claudin 5 protein levels in the PBLs of healthy controls (n = 13), OND controls (n = 14), T1D controls (n = 14), MS patients in remission (n = 11), and MS patients in relapse (n = 17). Box plot and data points display the relative protein levels determined by Western blot analysis. The relative intensity of claudin 1 or claudin 5 proteins compared to β-actin signals was determined by densitometry. Medians are depicted by horizontal bars.

Mentions: Following our observations that in vitro activation of leukocytes modulates TJP expression, ex vivo expression analysis was performed for PBLs from MS patients, OND, T1D and healthy control individuals. Claudin 1 protein levels were significantly higher in MS patients in relapse when compared to healthy controls (3.3-fold change, PB= 1.5 × 10−5), and to patients in remission (3.1-fold change, PB= 5.9 × 10−4; Fig. 3A). Claudin 1 protein levels were also higher in the T1D group when compared to healthy controls (2.4-fold change, PB = 7 × 10−5), but not in the OND group. The higher levels of claudin 1 observed in PBLs from the active autoimmune diseases, MS and T1D, is in line with the effects observed for the in vitro activations the expression of this protein in leukocytes. The RNA levels of CLDN1 could not be reliably analysed, because its transcript appeared to be very labile using the study’s standard RNA extraction procedures and in the quantitative RT-PCR assay (as opposed to the qualitative assay used in the preliminary screen) gave very variable results.


Tight junction proteins expression and modulation in immune cells and multiple sclerosis.

Mandel I, Paperna T, Glass-Marmor L, Volkowich A, Badarny S, Schwartz I, Vardi P, Koren I, Miller A - J. Cell. Mol. Med. (2012)

Expression levels of claudins 1 and 5 in leukocytes from MS and T1D. (A) Claudin 1 protein levels in PBLs of healthy controls (n = 13), OND controls (n = 14), T1D controls (n = 14), MS patients in remission (n = 13) and MS patients in relapse (n = 10). (B) The expression levels of CLDN5 RNA (2ΔCT) relative to reference genes were determined by real- time RT-PCR of PBL RNA derived from MS patients in relapse (n = 33), MS patients in remission (n = 39), and healthy controls (n = 18). (C) Claudin 5 protein levels in the PBLs of healthy controls (n = 13), OND controls (n = 14), T1D controls (n = 14), MS patients in remission (n = 11), and MS patients in relapse (n = 17). Box plot and data points display the relative protein levels determined by Western blot analysis. The relative intensity of claudin 1 or claudin 5 proteins compared to β-actin signals was determined by densitometry. Medians are depicted by horizontal bars.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3822847&req=5

fig03: Expression levels of claudins 1 and 5 in leukocytes from MS and T1D. (A) Claudin 1 protein levels in PBLs of healthy controls (n = 13), OND controls (n = 14), T1D controls (n = 14), MS patients in remission (n = 13) and MS patients in relapse (n = 10). (B) The expression levels of CLDN5 RNA (2ΔCT) relative to reference genes were determined by real- time RT-PCR of PBL RNA derived from MS patients in relapse (n = 33), MS patients in remission (n = 39), and healthy controls (n = 18). (C) Claudin 5 protein levels in the PBLs of healthy controls (n = 13), OND controls (n = 14), T1D controls (n = 14), MS patients in remission (n = 11), and MS patients in relapse (n = 17). Box plot and data points display the relative protein levels determined by Western blot analysis. The relative intensity of claudin 1 or claudin 5 proteins compared to β-actin signals was determined by densitometry. Medians are depicted by horizontal bars.
Mentions: Following our observations that in vitro activation of leukocytes modulates TJP expression, ex vivo expression analysis was performed for PBLs from MS patients, OND, T1D and healthy control individuals. Claudin 1 protein levels were significantly higher in MS patients in relapse when compared to healthy controls (3.3-fold change, PB= 1.5 × 10−5), and to patients in remission (3.1-fold change, PB= 5.9 × 10−4; Fig. 3A). Claudin 1 protein levels were also higher in the T1D group when compared to healthy controls (2.4-fold change, PB = 7 × 10−5), but not in the OND group. The higher levels of claudin 1 observed in PBLs from the active autoimmune diseases, MS and T1D, is in line with the effects observed for the in vitro activations the expression of this protein in leukocytes. The RNA levels of CLDN1 could not be reliably analysed, because its transcript appeared to be very labile using the study’s standard RNA extraction procedures and in the quantitative RT-PCR assay (as opposed to the qualitative assay used in the preliminary screen) gave very variable results.

Bottom Line: The tight junction proteins (TJPs) are major determinants of endothelial cells comprising physiological vascular barriers such as the blood-brain barrier, but little is known about their expression and role in immune cells.Interestingly, claudin 1 protein levels were elevated also in PBLs of patients with type 1 diabetes (T1D).Furthermore, a correlation between CLDN5 pre-treatment levels and clinical response phenotype to interferon-β therapy was detected.

View Article: PubMed Central - PubMed

Affiliation: Rappaport Faculty of Medicine and Research Institute, Technion-Israel Institute of Technology, Haifa, Israel.

Show MeSH
Related in: MedlinePlus