Vander Meyden M, Andersson U, Schmid I, Giorgi J, Kishimoto T, Martinez-Maza O; International Conference on AIDS.
Int Conf AIDS. 1992 Jul 19-24; 8: A61 (abstract no. PoA 2353).
UCLA School of Medicine.
OBJECTIVES: Interleukin 6 (IL6) is a multi-functional cytokine and the overproduction of which may play an important role in the immunogenesis of AIDS. We developed a rapid and sensitive method to detect cytoplasmic IL6 simultaneously with surface antigens. It is of particular interest for multiparametric analysis. METHODS: PBMC were collected from homosexual men (HIV- and HIV+), and healthy heterosexual men, ages 25-65 with an absolute CD4 count greater than 400mm3. Cells (2 x 10(6) cells/ml) were cultured with monensin (0.3 uM/ml, 1.5 h), permeabilized with 0.1% saponin, and fixed with periodate-lysin-paraformaldehyde (PLP). Cells were stained with anti-IL-6 (5 ug/ml) prior to staining for various cell surface antigens. Student's T test for multiple comparisons were done. RESULTS: The phenotypic characterization, based on morphology and two-color staining of the cytoplasmic-positive cells, indicates that monocytes are the predominant cell involved in IL6 production. In HIV positive donors (n = 15) 9.3% of monocytes produced IL6 compared to 3.9% (p less than 0.0001) of HIV negative donors (n = 15); and 4.4% (P less than 0.0008) in healthy controls (n = 15). Flow cytometry correlated (n = 15, r = .9526, p less than 0.003) with immunofluorescent microscopy. IL6 appeared as a local, polar accumulation in the cytoplasm in a juxtanuclear position identical to the Golgi apparatus. CONCLUSION: This method allows both early detection of cytokine production and multiparametric analysis. These results indicate that an elevated number of IL-6 containing monocytes are seen in PBMC isolated from HIV-infected donors.
Publication Types:
Keywords:
- AIDS Vaccines
- Acquired Immunodeficiency Syndrome
- Antigens, CD4
- Biological Markers
- Cytokines
- Cytoplasm
- Flow Cytometry
- HIV Infections
- HIV Seropositivity
- Homosexuality
- Humans
- Interleukin-6
- Male
- Monocytes
- Research
- immunology
Other ID:
UI: 102198376
From Meeting Abstracts