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Introduction: Flow cytometry allows simultaneous detection of surface and intracellular molecules on each cell., Objective: To describe a method for building up a harmonic multicolor panel with 11 flow cytometry parameters for phenotypic and functional analysis on CD8 + T lymphocytes., Materials and Methods: For the multicolor panel construction, we selected the molecules and titred conjugated antibodies with fluorochromes for CD3, CD8, CCR7, CD28, CD27, CD45RA, CD95 and CD127 determination in peripheral blood mononuclear cells (PBMC). To evaluate the panel, the conjugated antibodies were gradually added one by one and fluorescence minus one (FMO) test was performed. This method was applied to assess ex vivo subpopulations of T cells and the production of intracellular IFN ? , IL-2 and TNF a using polyclonal stimulation with enterotoxin B from Staphylococcus aureus (SEB) and antigen-specific cells with crude Trypanosoma cruzi antigen. Finally, the ex vivo CD8 + T lymphocyte subpopulations frequencies were analyzed in healthy individuals., Results: The evaluation of the selected molecules and conjugates did not show interference in the fluorescence signals and detection. The frequencies of CD8 + T cells evaluated were similar to the values reported in other studies. Additionally, we observed that the frequency of CD8 + T lymphocytes producing IFN ? , IL-2 and TNF a was higher 6 hours after culture with SEB and crude T. cruzi lysate., Conclusions: The method used for the construction of a multicolor panel allows obtaining frequencies of CD8 + T lymphocyte subpopulations corresponding to those reported in the literature.

In this paper we evaluated a new micro-flow injector for the determination of the concentrations of Cu, Mg and Zn in mononuclear blood cells. This device analyzed sample volumes in the order of microliters by flame atomic absorption spectrometry; it is inexpensive, and easy to build and to adapt to the conventional injector of the atomic absorption spectrophotometer. Detection limits of 106, 65 and 37 microg L(-1) for Cu, Mg and Zn were obtained, respectively. The percentages of recovery tests were found between 98 and 110%.

Background: To evaluate the diagnostic use of Polymerase Chain Reaction (PCR) for the detection of HIV-DNA., Methods: We have investigated HIV-DNA in peripheral blood mononuclear cells of 150 individuals: 50 seronegative without recognized factors to acquire HIV infection, 50 seronegative that showed risk habits and 50 seropositive. For amplification we have used primers SK38/SK39 which amplify a conserved region of 114 bp from HIV's gag gen. The detection of the amplificate was performed using solution hybridization with 32P-labelled SK 19, 6% polyacrylamide gel electrophoresis and autoradiography., Results: We have detected HIV-DNA at every seropositive patient (100%), 6 samples from seronegative individuals that showed risk habits and 1 sample from non-risk seronegative individuals., Conclusions: Polymerase chain reaction represents a diagnostic tool at HIV infection, specially for those patients where seroconversion can not be demonstrated by conventional methods.

We have investigated, by "in situ" hibridisation, the presence of hepatitis B virus (HBV) DNA in peripheral blood mononuclear cells (PBMC) from 45 patients with acute and chronic hepatic disorders directly related with HBV or with some seric HBV marker. Results has been related with serological markers and the different types of hepatopaties. The HBV-DNA was detected in PBMC more frequently in patients with hepatic alterations more prolongated (chronic active hepatitis, chronic persistent hepatitis and cirrhosis) than in acute hepatitis patients. It was not detected in any asymptomatic patient with HBV serological markers. As regards HBV serological markers, HBV-DNA was detected in PBMC in 8/11 HBsAg positive patients and in 11/34 HBsAg negative patients: 3 antiHBc positive, 5 antiHBc and antiHBs positive and 3 without conventional seric markers. The detection of HBV-DNA in antiHBc and/or antiHBs positive subjects means the virus may persist after recovery of infection and suggests PMBC could serve as additional reservoirs for reinfection of hepatocytes leading to a reactivation of the liver disease. Our results suggest that HBV infection of PBMC is a frequent event during HBV infection and can have important consequences fundamentally with respect to pathogenic mechanisms of HBV induced liver disease and to the transmission of the virus.