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An antibody microarray analysis of serum cytokines in neurodegenerative Parkinsonian syndromes.

Mahlknecht P, Stemberger S, Sprenger F, Rainer J, Hametner E, Kirchmair R, Grabmer C, Scherfler C, Wenning GK, Seppi K, Poewe W, Reindl M - Proteome Sci (2012)

Bottom Line: In a next step, results from the microarray experiment were individually validated by different immunoassays.Immunoassay validation confirmed a significant increase of median PDGF-BB levels in patients with PSP/CBS, MSA and PD and a decrease of median prolactin levels in PD.However, neither PDGF-BB nor prolactin were specific biomarkers to discriminate PSP/CBS, MSA, PD and controls.

View Article: PubMed Central - HTML - PubMed

Affiliation: Clinical Department of Neurology, Innsbruck Medical University, Innsbruck, Austria. Markus.Reindl@i-med.ac.at.

ABSTRACT

Background: Microarray technology may offer a new opportunity to gain insight into disease-specific global protein expression profiles. The present study was performed to apply a serum antibody microarray to screen for differentially regulated cytokines in Parkinson's disease (PD), multiple system atrophy (MSA), progressive supranuclear palsy (PSP) and corticobasal syndrome (CBS).

Results: Serum samples were obtained from patients with clinical diagnoses of PD (n = 117), MSA (n = 31) and PSP/CBS (n = 38) and 99 controls. Cytokine profiles of sera from patients and controls were analyzed with a semiquantitative human antibody array for 174 cytokines and the expression of 12 cytokines was found to be significantly altered. In a next step, results from the microarray experiment were individually validated by different immunoassays. Immunoassay validation confirmed a significant increase of median PDGF-BB levels in patients with PSP/CBS, MSA and PD and a decrease of median prolactin levels in PD. However, neither PDGF-BB nor prolactin were specific biomarkers to discriminate PSP/CBS, MSA, PD and controls.

Conclusions: In our unbiased cytokine array based screening approach and validation by a different immunoassay only two of 174 cytokines were significantly altered between patients and controls.

No MeSH data available.


Related in: MedlinePlus

Validation of seven differentially expressed cytokines in patients with PSP/CBS, MSA and controls in the screening cohort. Individual data points are shown as circles or triangles and horizontal bars indicate medians. In addition data are shown as box plots with medians indicated as horizontal bars with boxes. Groups were compared using the Kruskal-Wallis test and Dunn’s multiple comparison post-hoc test and overall p-values for comparison of PSP/CBS, MSA and combined controls or for comparison of PSP/CBS, MSA, HC and INC are shown in each figure. Ns = statistically not significant.
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Figure 2: Validation of seven differentially expressed cytokines in patients with PSP/CBS, MSA and controls in the screening cohort. Individual data points are shown as circles or triangles and horizontal bars indicate medians. In addition data are shown as box plots with medians indicated as horizontal bars with boxes. Groups were compared using the Kruskal-Wallis test and Dunn’s multiple comparison post-hoc test and overall p-values for comparison of PSP/CBS, MSA and combined controls or for comparison of PSP/CBS, MSA, HC and INC are shown in each figure. Ns = statistically not significant.

Mentions: Since the cytokine microarray analysis was performed using pooled samples from the screening cohort, we decided to validate the results of the microarray analysis by ELISA (MCP-4, prolactin, RANTES and IL-2RA) and by Flow-Cytomix assays (ICAM-1, leptin and PDGF-BB) applied on individual samples for seven of the 12 cytokines. These seven cytokines were selected based on the results from the microarray experiments, the availability of commercial available test kits and the amount of serum left. We used the same serum samples as shown in Table 1A and the results of these validation experiments are shown in Table 3 and Figure 2. However, only two (PDGF-BB and prolactin) of the seven cytokines were significantly different amongst patients (PSP/CBS, MSA and PD) and controls, whereas we could not confirm the cytokine microarray results for ICAM-1, IL-2RA, leptin, MCP-4 and RANTES (Figure 2). A separate analysis of both control groups, HC and INC, did not change the results.


An antibody microarray analysis of serum cytokines in neurodegenerative Parkinsonian syndromes.

Mahlknecht P, Stemberger S, Sprenger F, Rainer J, Hametner E, Kirchmair R, Grabmer C, Scherfler C, Wenning GK, Seppi K, Poewe W, Reindl M - Proteome Sci (2012)

Validation of seven differentially expressed cytokines in patients with PSP/CBS, MSA and controls in the screening cohort. Individual data points are shown as circles or triangles and horizontal bars indicate medians. In addition data are shown as box plots with medians indicated as horizontal bars with boxes. Groups were compared using the Kruskal-Wallis test and Dunn’s multiple comparison post-hoc test and overall p-values for comparison of PSP/CBS, MSA and combined controls or for comparison of PSP/CBS, MSA, HC and INC are shown in each figure. Ns = statistically not significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3539904&req=5

Figure 2: Validation of seven differentially expressed cytokines in patients with PSP/CBS, MSA and controls in the screening cohort. Individual data points are shown as circles or triangles and horizontal bars indicate medians. In addition data are shown as box plots with medians indicated as horizontal bars with boxes. Groups were compared using the Kruskal-Wallis test and Dunn’s multiple comparison post-hoc test and overall p-values for comparison of PSP/CBS, MSA and combined controls or for comparison of PSP/CBS, MSA, HC and INC are shown in each figure. Ns = statistically not significant.
Mentions: Since the cytokine microarray analysis was performed using pooled samples from the screening cohort, we decided to validate the results of the microarray analysis by ELISA (MCP-4, prolactin, RANTES and IL-2RA) and by Flow-Cytomix assays (ICAM-1, leptin and PDGF-BB) applied on individual samples for seven of the 12 cytokines. These seven cytokines were selected based on the results from the microarray experiments, the availability of commercial available test kits and the amount of serum left. We used the same serum samples as shown in Table 1A and the results of these validation experiments are shown in Table 3 and Figure 2. However, only two (PDGF-BB and prolactin) of the seven cytokines were significantly different amongst patients (PSP/CBS, MSA and PD) and controls, whereas we could not confirm the cytokine microarray results for ICAM-1, IL-2RA, leptin, MCP-4 and RANTES (Figure 2). A separate analysis of both control groups, HC and INC, did not change the results.

Bottom Line: In a next step, results from the microarray experiment were individually validated by different immunoassays.Immunoassay validation confirmed a significant increase of median PDGF-BB levels in patients with PSP/CBS, MSA and PD and a decrease of median prolactin levels in PD.However, neither PDGF-BB nor prolactin were specific biomarkers to discriminate PSP/CBS, MSA, PD and controls.

View Article: PubMed Central - HTML - PubMed

Affiliation: Clinical Department of Neurology, Innsbruck Medical University, Innsbruck, Austria. Markus.Reindl@i-med.ac.at.

ABSTRACT

Background: Microarray technology may offer a new opportunity to gain insight into disease-specific global protein expression profiles. The present study was performed to apply a serum antibody microarray to screen for differentially regulated cytokines in Parkinson's disease (PD), multiple system atrophy (MSA), progressive supranuclear palsy (PSP) and corticobasal syndrome (CBS).

Results: Serum samples were obtained from patients with clinical diagnoses of PD (n = 117), MSA (n = 31) and PSP/CBS (n = 38) and 99 controls. Cytokine profiles of sera from patients and controls were analyzed with a semiquantitative human antibody array for 174 cytokines and the expression of 12 cytokines was found to be significantly altered. In a next step, results from the microarray experiment were individually validated by different immunoassays. Immunoassay validation confirmed a significant increase of median PDGF-BB levels in patients with PSP/CBS, MSA and PD and a decrease of median prolactin levels in PD. However, neither PDGF-BB nor prolactin were specific biomarkers to discriminate PSP/CBS, MSA, PD and controls.

Conclusions: In our unbiased cytokine array based screening approach and validation by a different immunoassay only two of 174 cytokines were significantly altered between patients and controls.

No MeSH data available.


Related in: MedlinePlus