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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

Identification of serum biomarkers for the discrimination of movement disorders by antibody arrays in the screening cohort. (A) Representative picture of Raybiotech human cytokine antibody array showing the reactivity of pooled serum samples (10 PSP/CBS, 10 MSA, 20 PD and 30 controls) to arrays G series 2000 6, 7 and 8 (174 cytokines). Each protein was measured in duplicates. Signals were scanned with a GenePix 4000B scanner. Blue boxes: positive controls (upper left corner, high intense spots). Red box, negative controls (upper left and lower right corner, no spots). Purple boxes, internal controls IC1, IC and IC3 (lower right corner, spots with three different intensities). White and green colored boxes indicate the location of the detection of two proteins that were significantly different in both the microarray and validation experiment (white = PDGF-BB and green = prolactin). (B) Normalized array data of the 174 cytokines were analyzed by SAM to detect differences in their concentrations between pooled serum samples (PSP/CBS, one pool of 10 samples with three replicates; MSA, one pool of 10 samples with three replicates each; PD, two pools of 10 samples with two replicates each; and controls, three pools of 10 samples with two replicates each). The relative concentrations of the 12 cytokines that obtained a significant score (q-value <0.001%) are shown in a “heatmap”. Low concentrations are shown in blue, median concentrations in black and high concentrations in yellow.
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Figure 1: Identification of serum biomarkers for the discrimination of movement disorders by antibody arrays in the screening cohort. (A) Representative picture of Raybiotech human cytokine antibody array showing the reactivity of pooled serum samples (10 PSP/CBS, 10 MSA, 20 PD and 30 controls) to arrays G series 2000 6, 7 and 8 (174 cytokines). Each protein was measured in duplicates. Signals were scanned with a GenePix 4000B scanner. Blue boxes: positive controls (upper left corner, high intense spots). Red box, negative controls (upper left and lower right corner, no spots). Purple boxes, internal controls IC1, IC and IC3 (lower right corner, spots with three different intensities). White and green colored boxes indicate the location of the detection of two proteins that were significantly different in both the microarray and validation experiment (white = PDGF-BB and green = prolactin). (B) Normalized array data of the 174 cytokines were analyzed by SAM to detect differences in their concentrations between pooled serum samples (PSP/CBS, one pool of 10 samples with three replicates; MSA, one pool of 10 samples with three replicates each; PD, two pools of 10 samples with two replicates each; and controls, three pools of 10 samples with two replicates each). The relative concentrations of the 12 cytokines that obtained a significant score (q-value <0.001%) are shown in a “heatmap”. Low concentrations are shown in blue, median concentrations in black and high concentrations in yellow.

Mentions: We have applied a screening approach with human cytokine antibody arrays using pooled serum samples from patients with PSP/CBS, MSA, PD and age and sex-matched controls (CTRL) to identify putative serum biomarkers for these diseases (Table 1A). In order to exclude an effect of age-related non-neurological diseases on serum cytokine levels we have not only included 63 age and sex matched healthy blood donors (HC), but also 36 patients with internal diseases (INC) in our control group. Figure 1 shows a heatmap of the microarrays for the four groups of patients and controls.


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)

Identification of serum biomarkers for the discrimination of movement disorders by antibody arrays in the screening cohort. (A) Representative picture of Raybiotech human cytokine antibody array showing the reactivity of pooled serum samples (10 PSP/CBS, 10 MSA, 20 PD and 30 controls) to arrays G series 2000 6, 7 and 8 (174 cytokines). Each protein was measured in duplicates. Signals were scanned with a GenePix 4000B scanner. Blue boxes: positive controls (upper left corner, high intense spots). Red box, negative controls (upper left and lower right corner, no spots). Purple boxes, internal controls IC1, IC and IC3 (lower right corner, spots with three different intensities). White and green colored boxes indicate the location of the detection of two proteins that were significantly different in both the microarray and validation experiment (white = PDGF-BB and green = prolactin). (B) Normalized array data of the 174 cytokines were analyzed by SAM to detect differences in their concentrations between pooled serum samples (PSP/CBS, one pool of 10 samples with three replicates; MSA, one pool of 10 samples with three replicates each; PD, two pools of 10 samples with two replicates each; and controls, three pools of 10 samples with two replicates each). The relative concentrations of the 12 cytokines that obtained a significant score (q-value <0.001%) are shown in a “heatmap”. Low concentrations are shown in blue, median concentrations in black and high concentrations in yellow.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: Identification of serum biomarkers for the discrimination of movement disorders by antibody arrays in the screening cohort. (A) Representative picture of Raybiotech human cytokine antibody array showing the reactivity of pooled serum samples (10 PSP/CBS, 10 MSA, 20 PD and 30 controls) to arrays G series 2000 6, 7 and 8 (174 cytokines). Each protein was measured in duplicates. Signals were scanned with a GenePix 4000B scanner. Blue boxes: positive controls (upper left corner, high intense spots). Red box, negative controls (upper left and lower right corner, no spots). Purple boxes, internal controls IC1, IC and IC3 (lower right corner, spots with three different intensities). White and green colored boxes indicate the location of the detection of two proteins that were significantly different in both the microarray and validation experiment (white = PDGF-BB and green = prolactin). (B) Normalized array data of the 174 cytokines were analyzed by SAM to detect differences in their concentrations between pooled serum samples (PSP/CBS, one pool of 10 samples with three replicates; MSA, one pool of 10 samples with three replicates each; PD, two pools of 10 samples with two replicates each; and controls, three pools of 10 samples with two replicates each). The relative concentrations of the 12 cytokines that obtained a significant score (q-value <0.001%) are shown in a “heatmap”. Low concentrations are shown in blue, median concentrations in black and high concentrations in yellow.
Mentions: We have applied a screening approach with human cytokine antibody arrays using pooled serum samples from patients with PSP/CBS, MSA, PD and age and sex-matched controls (CTRL) to identify putative serum biomarkers for these diseases (Table 1A). In order to exclude an effect of age-related non-neurological diseases on serum cytokine levels we have not only included 63 age and sex matched healthy blood donors (HC), but also 36 patients with internal diseases (INC) in our control group. Figure 1 shows a heatmap of the microarrays for the four groups of patients and controls.

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