Pandolfi F, Cafaro A, Polidori V, Oliva A, Liberatore D, Sacco G, Giovannetti A, Kumick JT, Aluti F; International Conference on AIDS.
Int Conf AIDS. 1992 Jul 19-24; 8: A60 (abstract no. PoA 2347).
Dept of Clinical Immunology, Univ. of Rome.
In comparison to normal controls, peripheral blood mononuclear cells (PBMC) from HIV-infected patients have an increased mortality rate (MR) when incubated in vitro in a survival medium (RPMI 1640 supplemented with 10% FCS and antibiotics). PBMC were isolated from 49 HIV+ patients and cultured in a CO2 incubator at 37 degrees C for 3 days in RPMI 1640. Cells were then collected, stained with 0.5 micrograms/ml ethidium bromide and analyzed with a flow cytometer. MR was calculated as follows: TABULAR DATA, SEE ABSTRACT VOLUME. Overall MR was 30.5% in HIV+ patients in comparison to 11.8% in normal controls (P = 0.002). Differences in MR between patients and controls were higher in 25 patients in Stage IV (CDC classification) (P = 0.001) and in 24 patients in Stages II and III (P = 0.007, in comparison to controls). MR was higher in 38 patients with CD4+ less than 400/cu mm in comparison to patients with greater than 400 CD4+ (33.5 vs 20.9%, P = 0.037, both being significantly different from controls). Addition to the culture of calcium ionophore increased by more than 2 fold the MR in 6 HIV+ patients. We have previously shown (J. exp. Med., 172, 1873, 1990) that fibroblast conditioned medium (FCM) can preserve viability, without significant activation, of human lymphocytes in vitro. We therefore tested the ability of two FCM (-1 and -2, derived from COS and RAT-1 cell lines) and a panel of cloned cytokines (GM-CSF, IL-2, IL-6, IL-7) to reduce spontaneous MR in HIV+ PBMC. Results indicate that FCM-1 and IL-2 significantly reduced MR in HIV+ patients (MR were 20.7 and 18.5%, P = 0.002 and 0.0001, respectively). However, cellular proliferation can account for increased survival of cells in IL-2 as shown by 3H incorporation. In the group of patients with less than 400 CD4/cu mm, both FCM-1 and -2 significantly reduced MR (P = 0.006 and 0.04, respectively). High or low molecular weight DNA was extracted from fresh PBMC and cells cultured for 3 days in 8 and 12 HIV+ cases. In all cases DNA degradation was observed after culture. In only 3 patients bands possibly related to an apoptotic mechanism of death were observed. DNA extracted from the WEHI 231 cell line activated with anti-IgM was used as positive control of apoptosis. Our data suggest that FCM decrease in vitro spontaneous MR in PBMC isolated from HIV+ patients.
Publication Types:
Keywords:
- AIDS Vaccines
- Acquired Immunodeficiency Syndrome
- Animals
- Antigens, CD4
- Apoptosis
- Cell Line
- Cytokines
- Fibroblasts
- Flow Cytometry
- Granulocyte-Macrophage Colony-Stimulating Factor
- Greece
- HIV Infections
- HIV Seropositivity
- Humans
- In Vitro
- Interleukin-2
- Rats
- immunology
Other ID:
UI: 102198368
From Meeting Abstracts