Granulysin Expression in Lymphocytes that Populate the Peripheral Blood and the Myocardium after an Acute Coronary Event.

We aimed to analyse granulysin (GNLY)-mediated cytotoxicity in the peripheral blood of patients with non-ST-segment elevation myocardial infarction (NSTEMI) treated with anti-ischaemic drug therapy. Thirty-nine NSTEMI patients with a median age of 70 years and 28 age-matched healthy subjects were enrolled in this study. On day 7 after MI, the number of GNLY(+) lymphocytes in the peripheral blood increased approximately six-fold of that in the healthy subjects, measured by flow cytometry. On day 14, the number of GNLY(+) cells significantly decreased in T, NKT, and both CD56(+dim) and CD56(+bright) NK subsets. GNLY(+) CD3(+) and GNLY(+) CD56(+) cells infiltrated central zone of myocardial infarction (MI). In persons who died in the first week after MI, GNLY(+) cells were found within accumulation of apoptotic leucocytes and reached the apoptotic cardiomyocytes in border MI zones probably due to the influence of interleukin-15 in peri-necrotic cardiomyocytes, as it is was shown by immunohistology. By day 28, the percentage of GNLY(+) lymphocytes in peripheral blood returned to the levels similar to that of the healthy subjects. Anti-GNLY mAb decreased apoptosis of K562 targets using peripheral blood NK cells from days 7 and 28 after MI, while in assays using cells from days 1 and 21, both anti-GNLY and anti-perforin mAbs were required to significantly decrease apoptosis. Using NK cells from day 14, K562 apoptosis was nearly absent. In conclusion, it seems that GNLY(+) lymphocytes, probably attracted by IL-15, not only participate partially in myocardial cell apoptosis, but also hasten resolution of cardiac leucocyte infiltration in patients with NSTEMI.

Perforin-mediated cytotoxicity in non-ST elevation myocardial infarction.

The aim of this investigation was to examine the role of perforin (P)-mediated cytotoxicity in the dynamics of tissue damage in patients with non-ST-segment elevation myocardial infarction (NSTEMI) treated with anti-ischaemic drugs. We enrolled 48 patients with NSTEMI in this study [age, 71.5 years; 61.5/76 (median, 25th/75th percentiles)]. The percentage of total peripheral blood P(+) lymphocytes was elevated owing to the increased frequency of P(+) cells within natural killer (NK) subsets, T and NKT cells in patients on day 1 after NSTEMI when compared with healthy controls. Positive correlations were found between cardiac troponin I plasma concentrations and the frequency of P(+) cells, P(+) T cells, P(+) NK cells and their CD56(+dim) and CD56(+bright) subsets during the first week after the NSTEMI. The expression of P in NK cells was accompanied by P-mediated cytotoxicity against K-562 targets at all days examined, except day 21, when an anti-perforin monoclonal antibody did not completely abolish the killing. The percentage of P(+) T cells, P(+) NKT cells and P(+) NK subsets was the highest on the day 1 after NSTEMI and decreased in the post-infarction period. CD56(+) lymphocytes were found in damaged myocardium, suggesting their tissue recruitment. In conclusion, patients with NSTEMI have a strong and prolonged P-mediated systemic inflammatory reaction, which may sustain autoaggressive reactions towards myocardial tissue during the development of myocardial infarction.

Biochemical and cytochemical comparison of surface membranes from normal and dystrophic chickens.

Cytochemical and biochemical characteristics of the surface membrane components of avian dystrophic muscle were examined. A Mg2+- or Ca2+-activated ("basic") adenosine triphosphate (ATPase) was localized cytochemically in fixed, intact dystrophic muscle slices in a medium containing Mg2+ or Ca2+, adenosine triphosphate (ATP), and 1 microM free Pb2+ to capture enzymatically released phosphate ions. Electron-dense staining precipitates were found to be associated with the plasmalemma and its tortuous invaginations, and the transverse components of the T-system membrane and its associated proliferated networks. Enzymatic analysis of microsomal fractions isolated from 7-day-old and 90-day-old normal and dystrophic muscle showed a complex behavior. Specific activity of "basic" ATPase decreased with maturity in normal and dystrophic animals. The specific activities of the surface membrane associated enzymes, leucyl beta-naphthylamidase, adenylate cyclase, and guanylate cyclase, remained at various elevated levels in the mature dystrophic animals, in contrast to the normal muscle, which showed decreases in the specific activity of all three enzymes with maturation. The persistent high levels in some but not all enzyme activities in 90-day-old dystrophic muscle indicates a complicated developmental pattern in the dystrophic chicken muscle.