Cytokine profiles in HIV-1 subtype CRF01_AE infected individuals with different rates of diseases progression: a multiplex immunoassay.

OBJECTIVE: Cytokines play an important role in controlling the homeostasis of the immune system and contribute to the pathogenesis of HIV infection. The measurement soluble cytokines in plasma of HIV-1 infected individuals with different rates of disease progression may provide additional information to complement prognostic markers and understand disease process. The aim of the present study was to determine the cytokine profiles in plasma of Thai HIV-1 CRFO1_AE infected individuals with different rates of disease progression by using a multiplex system for simultaneous detection of 7 cytokines. MATERIAL AND METHOD: The authors used a multiplex immunoassay method to measure 7 cytokines (IL-2, IL-4, IL-6, IL-7, IL-10, IL-15 and IFN-gamma) in plasma of 23 progressors (PRs; symptomatic or AIDS within 5 years and CD4+ < 200/mm3), 23 slower progressors (SPs; asymptomatic more than 5 years and CD4+ > 350/mm3) and 23 normal healthy individuals. RESULTS: Both PRs and SPs demonstrated significantly higher levels of IL-7, IL-10 and IFN-gamma than healthy controls (p < 0.05). No significant difference in IL-6 between SPs and healthy controls but significant difference between RPs and controls were found. Furthermore, PRs showed significantly higher levels of plasma IL-6 (p = 0.001), IL-7 (p = 0.016), IL-10 (p < 0.001) and IFN-gamma (p = 0.026) than SPs. No significant difference in IL-2, IL-4 and IL-15 was found among 3 groups (PRs, SPs and healthy control). CONCLUSION: These results suggested that a Th1 to Th2 cytokine switch did not occur. However, the measurements of plasma levels of cytokines could be used for predicting disease progression.

A whole blood lymphocyte proliferation assay in healthy Thais: comparison of heparinized blood and acid citrate dextrose blood.

The lymphocyte proliferation assay (LPA) is a technique to determine T-lymphocyte functions in vitro. The standard LPA using peripheral blood mononuclear cells (PBMC) separated from heparinized blood requires a large blood sample, time consuming and expensive. It is more useful if acid citrate dextrose (ACD) blood could be used not only for LPA but also for other purposes. To determine whether whole blood composing between heparinized blood and ACD blood could be substituted for standard LPA using PBMC. Heparinized and ACD blood of 35 healthy Thai blood donors were studied herein. PBMC separated by density gradient centrifugation and diluted heparinized and ACD blood were used to test and compare for lymphoproliferative responses to phytohemagglutinin (PHA), pokeweed mitogen (PWM), and tetanus toxoid. A stimulation index (SI) for each mitogen or antigen was calculated. All Thai blood donors demonstrated positive proliferative responses to PHA and PWM by using PBMC and whole blood culture assays from both heparinized and ACD blood. However, the difference in the frequency of positive proliferative responses to tetanus toxoid by using PBMC and whole blood culture assays was significant. Nevertheless, no significant difference in frequency of positive responses to tetanus toxoid between heparinized and ACD blood was observed. This results suggested that no significant difference between using heparinized and ACD blood in standard LPA using PBMC. However, the whole blood LPA for measuring mitogen induced lymphoproliferation could be substituted for standard LPA from heparinized andACD blood. Whole blood LPA is easy, rapid, and more cost effective than PBMC culture assay. Thus, it would be applicable in a clinical laboratory as well as in research setting.