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TNF alpha production by whole blood from HIV-1 infected patients
1Laboratoire de Virologie, Bât. IRFPPS, CHU, Lille, France.
Insights
This study developed a rapid whole blood assay to measure tumor necrosis factor alpha (TNF-α) in HIV-1 patients. While average TNF-α levels were similar to controls, individual variations in HIV-1 patients suggest potential for monitoring disease progression.
Area of Science:
- Immunology
- Virology
- Biochemistry
Background:
- Cytokine production by immunocompetent cells is crucial in immune responses.
- Whole blood (WB) culture offers a physiologically relevant model for studying cytokine production in vitro.
- Tumor necrosis factor alpha (TNF-α) plays a significant role in immune regulation and disease pathogenesis.
Purpose of the Study:
- To develop and validate a one-step whole blood assay for measuring TNF-α production in HIV-1 positive individuals.
- To assess TNF-α production in HIV-1 patients at different disease stages compared to healthy controls.
- To evaluate the variability of TNF-α production over time in HIV-1 patients.
Main Methods:
- A one-step procedure combining whole blood stimulation (LPS and PHA) and TNF-α measurement was used.
- Small volumes (25 microliters) of heparinized whole blood were analyzed.
- 30 HIV-1 positive patients (CDC-staged) and 12 healthy seronegative subjects were studied.
- Sequential blood collection over 5 months was performed for variability assessment.
Main Results:
- Mean TNF-α levels in HIV-1 patients were not statistically different from healthy controls.
- Some HIV-1 patients exhibited TNF-α levels significantly higher or lower than controls.
- Individual TNF-α production showed limited variation in healthy controls but significant fluctuations in most HIV-1 patients over time.
Conclusions:
- The developed whole blood assay can detect abnormal TNF-α production in HIV-1 positive individuals.
- This method can be a valuable tool for investigating the role of cytokines in HIV-1 infection outcomes.
- Monitoring TNF-α levels may provide insights into disease progression and immune status in HIV-1 patients.
Abstract:
Production of cytokines by immunocompetent cells in vitro may be assessed after stimulation with polyclonal activators. Because it mimics the natural environment, diluted whole blood (WB) culture may be the most appropriate milieu in which to study cytokine production in vitro. We tested TNF alpha production by small volume of whole blood (25 microliters) from HIV-1 positive patients by using a one-step procedure that combines WB stimulation with LPS and PHA and cytokine measurement. We studied 30 patients without secondary infection or at distance of secondary infection staged according to the classification proposed by the CDC and 12 healthy seronegative subjects. The mean values of TNF alpha from patients were not statistically different from those from normal controls however in certain patients at different stages of the disease higher values than the mean +2 SD of controls and lower values than the mean -2 SD of controls were obtained. Heparinized blood from 5 control subjects had been collected sequentially during a period of 5 months. The individual variation of TNF alpha production were limited for all these individuals. For each of 6 HIV-1 patients, whole blood samples were collected sequentially during a period of 5 months and in most of patients large variations of levels of TNF alpha were observed from one sample to another one. Our method can detect abnormal cytokine production in HIV-1 positive individuals and can become a useful tool to investigate the role of cytokines in the outcome of HIV-1 infection.