Cytokine production by stimulated HD helper T lymphocytes is not altered compared to controls.

<p>CD4⁺ helper T lymphocytes were isolated from HD and control peripheral blood using magnetic cell sorting and stimulated with 5 μg/ml anti-CD3 and 2 μg/ml anti-CD28 antibodies. Supernatants were collected after 48 h and cytokine profiling was carried out. No significant differences were seen in the levels of any of the cytokines produced by HD and control CD4⁺ T lymphocytes after normalisation to total protein levels. Data show mean concentrations ± SEM. Statistical analysis was carried out using two-tailed unpaired student’s t tests.</p

The top ten gene changes in unstimulated Huntington’s disease helper T lymphocytes compared to controls.

<p>Data presented as fold change calculated from ΔΔ-CT values, unpaired two-tailed t-test used as statistical method.</p><p>The top ten gene changes in unstimulated Huntington’s disease helper T lymphocytes compared to controls.</p

The frequency of T lymphocyte subsets does not differ between HD and control peripheral blood.

<p>The frequencies of a range of T lymphocyte subsets in the isolated PBMC populations were measured by flow cytometry using antibodies to CD3⁺ T lymphocytes, CD3⁺ CD4⁺ helper T lymphocytes and CD3⁺ CD8⁺ cytotoxic T lymphocytes. Within the CD3⁺ CD4⁺ helper T lymphocyte population, the T_h1 and T_h2 subtypes were analysed using antibodies to CXCR3 and CCR4 respectively. No significant differences were seen between HD and control for any of the cell types that were analysed. Statistical analysis was carried out using two-tailed unpaired student’s t tests.</p

The number of divisions which proliferating CD3⁺ T lymphocytes undergo is not affected in HD.

<p>PBMCs were stained with the intracellular dye CFSE and the number of divisions which proliferating CD3⁺ T lymphocytes underwent <b>(A)</b> in the absence of stimulation <b>(B)</b> with 5 μg/ml anti-CD3 and 2 μg/ml anti-CD28 antibody stimulation and <b>(C)</b> with 10 μg/ml PHA-P stimulation was measured after 72, 96 and 120 h by flow cytometry. No significant differences were seen between HD and control for any of the experimental time points or conditions. Statistical analysis was carried out using two-tailed unpaired student’s t tests with a Holm-Šídák correction (alpha = 0.05) for multiple comparisons.</p

The percentage of proliferating CD3⁺ T lymphocytes is not affected in HD compared to control.

<p>PBMCs were stained with CFSE and the percentage of CD3⁺ T lymphocytes from the initial culture which divided <b>(A)</b> in the absence of stimulation <b>(B)</b> with 5 μg/ml anti-CD3 and 2 μg/ml anti-CD28 antibody stimulation and <b>(C)</b> with 10 μg/ml PHA-P stimulation was measured after 72, 96 and 120 h by flow cytometry. No significant differences were seen between HD and control for any of the experimental time points or conditions. Statistical analysis was carried out using two-tailed unpaired student’s t tests with a Holm-Šídák correction (alpha = 0.05) for multiple comparisons.</p

Cerebrospinal Fluid Inflammatory Biomarkers Reflect Clinical Severity in Huntington’s Disease

<div><p>Introduction</p><p>Immune system activation is involved in Huntington’s disease (HD) pathogenesis and biomarkers for this process could be relevant to study the disease and characterise the therapeutic response to specific interventions. We aimed to study inflammatory cytokines and microglial markers in the CSF of HD patients.</p><p>Methods</p><p>CSF TNF-α, IL-1β, IL-6, IL-8, YKL-40, chitotriosidase, total tau and neurofilament light chain (NFL) from 23 mutation carriers and 14 healthy controls were assayed.</p><p>Results</p><p>CSF TNF-α and IL-1β were below the limit of detection. Mutation carriers had higher YKL-40 (p = 0.003), chitotriosidase (p = 0.015) and IL-6 (p = 0.041) than controls. YKL-40 significantly correlated with disease stage (p = 0.007), UHDRS total functional capacity score (r = -0.46, p = 0.016), and UHDRS total motor score (r = 0.59, p = 4.5*10⁻⁴) after adjustment for age.</p><p>Conclusion</p><p>YKL-40 levels in CSF may, after further study, come to have a role as biomarkers for some aspects of HD. Further investigation is needed to support our exploratory findings.</p></div

Relationship between CSF concentration of YKL-40 and measures of disease progression and clinical severity.

<p>a, disease stage; b, UHDRS total functional score; c, UHDRS total motor score. The p-values for partial Pearson’s correlation coefficient adjusted for age are shown.</p

Comparison in CSF levels of inflammatory molecules between 14 healthy controls versus 23 gene expansion carriers.

<p>The p-values for unpaired two-tailed t-test (Il-8) or Wilcoxon log-rank test (YKL-40, chitotriosidase, IL-6) are shown.</p

Characteristics of included participants by disease stage.

<p>Characteristics of included participants by disease stage.</p

Results of multiple linear regression of model parameters on total motor score (TMS)—higher scores indicate worse motor problems.

<p>Results of multiple linear regression of model parameters on total motor score (TMS)—higher scores indicate worse motor problems.</p