The effects of overnight shipping on PBMC yield and survival.

<p>The cell number of blood derived PBMC was determined based on Trypan blue exclusion. A) Blood was collected in S-Monovette (Sarsted) or K2E Vacutainer (BD) tubes from 4 donors on site or at another location. Shipped samples were transported in the collection tube. PBMC yield was calculated per volume (10⁶ living PBMC/ml blood). B) Yield (10⁶ living PBMC/ml blood) is shown for PBMC derived from non-shipped vs. shipped blood (n = 9) prior to freezing. C) Viability (% of living cells/total cell count) is shown for non-shipped vs. shipped samples (n = 9), prior to freezing. D) Post-cryopreservation recovery is calculated for thawed samples (non-shipped vs. shipped, n = 9) and is provided as percentage of living cells from 10⁷ total (frozen amount/vial).</p

Additional file 2: of Human central nervous system astrocytes support survival and activation of B cells: implications for MS pathogenesis

Figure S1. Confirming activation of human astrocytes. Astrocytes were cultured for 24 h and were either left unstimulated or were stimulated with IFNγ (10 ng/ml) and IL-1β (10 ng/ml). After 24 h, the astrocytes were washed thoroughly and fresh medium was added. After an additional 24 h in culture, at which time cultures were imaged and supernatants were collected for subsequent measurement of astrocyte-secreted IL-6 by ELISA. Compared to unstimulated astrocytes (a), stimulated astrocytes exhibited activated morphology (b) and significantly-enhanced production of IL-6 (c; p = 0.0016; paired t-test). (TIFF 3951 kb

Additional file 3: of Human central nervous system astrocytes support survival and activation of B cells: implications for MS pathogenesis

Figure S2. Effects of astrocytes cytokine neutralization on B cell survival and activation. B cells from HC were either cultured alone, or with stimulated astrocyte conditioned-medium (ACM), or with ACM pre-treated with neutralizing antibodies to IL-6 (a, b; anti-IL6: aIL-6), IL-15 (c, d; anti-IL-15: aIL-15) or BAFF (e, f; anti-BAFF: aBAFF); or pre-treated with corresponding isotype control antibodies. After 2 days of culture B cell viability was assessed using ANNEXIN V and 7AAD staining, and CD86 expression was measured by flow cytometry (representative experiment). (TIFF 4226 kb

Supplementary Appendix, MSJ779952_supplementary_appendix – Detection and clinical correlation of leukocortical lesions in pediatric-onset multiple sclerosis on multi-contrast MRI

<p>Supplementary Appendix, MSJ779952_supplementary_appendix for Detection and clinical correlation of leukocortical lesions in pediatric-onset multiple sclerosis on multi-contrast MRI by Josefina Maranzano, Christine Till, Haz-Edine Assemlal, Vladimir Fonov, Robert Brown, David Araujo, Julia O’Mahony, E Ann Yeh, Amit Bar-Or, Ruth Ann Marrie, Louis Collins, Brenda Banwell, Douglas L Arnold and Sridar Narayanan in Multiple Sclerosis Journal</p

Evaluating Soluble EMMPRIN as a Marker of Disease Activity in Multiple Sclerosis: Studies of Serum and Cerebrospinal Fluid

<div><p><i>E</i>xtracellular <i>m</i>atrix <i>m</i>etallo<i>pr</i>oteinase <i>in</i>ducer (EMMPRIN, CD147) is an inducer of matrix metalloproteinases and has roles in leukocyte activation and migration. We reported previously that in MS and its animal model, experimental autoimmune encephalomyelitis, cell surface-associated EMMPRIN was significantly elevated in leukocytes around inflammatory perivascular cuffs in the CNS. In this study we report that activated T-cells can secrete soluble form of EMMPRIN (sEMMPRIN) upon activation. As sEMMPRIN is also present in biological fluids, we determined whether sEMMPRIN is altered in the CSF and sera of MS subjects. Sera from individuals without neurological conditions served as controls, while CSFs collected from subjects undergoing discectomy, and without evidence of CNS pathology, were used as a comparator group. We found that serum levels of sEMMPRIN from clinically stable MS patients or other inflammatory conditions did not differ from control subjects. Paired serum and CSF samples demonstrated poor correlation of sEMMPRIN. Interestingly, sEMMPRIN levels were approximately 60% higher in CSFs compared to sera. sEMMPRIN CSF levels were significantly higher in secondary progressive compared to primary progressive subjects. Thus we conclude that measurement of sEMMPRIN in serum is not informative for disease activity in MS. The differential expression of sEMMPRIN in the CSF of primary and secondary progressive MS invites hypotheses of the still undefined roles of EMMPRIN in the CNS.</p></div

sEMMPRIN in sera was not altered in different inflammatory conditions.

<p>sEMMPRIN in sera from HC (healthy controls) and patients with CD (Crohn’s disease), UC (ulcerative colitis), T1D (Type 1 diabetes) or RRMS were measured by ELISA. The number of subjects is shown in parentheses. The scatter plot shows that sEMMPRIN levels did not significantly differ between the groups (one-way ANOVA with Tukey’s post hoc test). Values are represented in pg/ml, with each entry representing a single subject performed in duplicate, and data is represented as mean ± SD.</p

Supplementary Material 1, MSJ779952_supplementary_material_1 – Detection and clinical correlation of leukocortical lesions in pediatric-onset multiple sclerosis on multi-contrast MRI

<p>Supplementary Material 1, MSJ779952_supplementary_material_1 for Detection and clinical correlation of leukocortical lesions in pediatric-onset multiple sclerosis on multi-contrast MRI by Josefina Maranzano, Christine Till, Haz-Edine Assemlal, Vladimir Fonov, Robert Brown, David Araujo, Julia O’Mahony, E Ann Yeh, Amit Bar-Or, Ruth Ann Marrie, Louis Collins, Brenda Banwell, Douglas L Arnold and Sridar Narayanan in Multiple Sclerosis Journal</p