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Published on: April 27, 2011
Biochip array-based analysis of plasma cytokines in HIV patients with immunological and virological discordance
N Sachdeva1, H S Yoon, K Oshima
1Laboratory for Clinical and Biological Studies, University of Miami-Miller School of Medicine, Miami, FL 33136, USA.
Insights
This study found that HIV patients with discordant immunological and virological profiles have higher levels of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF). Biochip array technology offers a promising method for simultaneous cytokine detection in limited samples.
Area of Science:
- Immunology
- Biotechnology
- Virology
Background:
- Cytokine assessment is crucial for understanding disease progression and guiding treatment strategies.
- Antibody-based protein arrays enable simultaneous detection of multiple analytes from small sample volumes.
- Investigating cytokine profiles in HIV patients with discordant immunological and virological responses is essential.
Purpose of the Study:
- To compare plasma cytokine profiles between HIV patients with discordant and concordant immunological and virological status.
- To evaluate the utility of a biochip array system for multiplex cytokine analysis in HIV research.
- To identify specific cytokines or growth factors that differ between discordant and concordant HIV-infected individuals.
Main Methods:
- A biochip array system was used to measure 12 cytokines and growth factors in plasma samples.
- Plasma specimens from 110 HIV patients (55 discordant, 55 concordant) and 22 healthy controls were analyzed.
- Enzyme-linked immunosorbent assay (ELISA) was employed to confirm significant findings from the biochip array.
Main Results:
- Discordant HIV patients exhibited significantly higher plasma levels of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) compared to concordant patients.
- HIV patients, overall, showed elevated levels of TNF-alpha, MCP-1, and VEGF compared to healthy controls.
- ELISA confirmed the higher levels of VEGF and EGF in discordant patients, validating the biochip array results.
Conclusions:
- The biochip array system is a reliable and comparable technology to ELISA for cytokine quantitation.
- This multiplexing technology holds significant potential for analyzing multiple cytokines in limited specimens, particularly in HIV research.
- Elevated VEGF and EGF levels in discordant HIV patients may indicate specific disease mechanisms or therapeutic targets.
Abstract:
Assessment of cytokines in body fluids or cells provides important information in understanding the disease process and designing treatment strategies. Recent introduction of antibody-based protein arrays have provided investigators simultaneous and specific detection of multiple analytes in a single sample using minimum volumes. In this study, we used a biochip array system capable of measuring 12 cytokines and growth factors (IL-2, IL-4, IL-6, IL-8, IL-10, IL-1alpha, IL-1beta, IFN-gamma, TNF-alpha, monocyte chemoattractant protein-1 (MCP-1), vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF)) in HIV patients with immunological and virological discordance (discordant) to find out differences if any, in their plasma cytokine profiles when compared with concordant HIV-infected individuals. A sandwich chemiluminescent assay was performed with plasma specimens of 110 HIV patients (55 discordant, 55 concordant) and 22 normal healthy individuals followed by enzyme-linked immunosorbent assay (ELISA) to the confirm levels of cytokines and growth factors that showed significant differences in the two groups. The discordant HIV patients showed significantly higher levels of plasma VEGF (P = 0.001) and EGF (P = 0.034) levels when compared with concordant patients. Overall, the patients showed significantly higher levels of TNF-alpha, MCP-1 and VEGF when compared with the normal healthy controls (P < 0.05). ELISA for VEGF (P < 0.001) and EGF (P = 0.004) confirmed the comparison obtained with biochip array, between the discordant and concordant patients. The results of cytokine quantitation by biochip array and ELISA confirmed that this technology is not only comparable but also has a good potential in the future applications involving measurement of multiple cytokines with limiting specimens.
