The effect of HIV-1 infection on the lipid fatty acid content in the membrane of cultured lymphocytes.

Elevations in the levels of unsaturated fatty acids (FAs) in membrane lipids lead to an increase in cell membrane fluidity and may also be involved in cell fusion and death through the loss of normal membrane function and integrity. Since the infection of susceptible cells with HIV leads to cell fusion and subsequent loss of viability, the present study was undertaken to see whether HIV infection can alter the relative content of unsaturated FAs in the host cell membrane and to determine whether this change correlates with cell death. Peripheral lymphocytes (PBLs) of a healthy donor and two CD4+ cell lines were chosen: MT-4, which is killed following HIV infection, with significant cell death being observed 5 days postinfection, and H9 which is not killed. Measurements of FA content of the two cell lines and PBLs, either before or at 6, 24, and 48 h after infection, showed a significant rise in the concentration of unsaturated FAs followed by a drop in the concentration of saturated FAs in the MT-4 cell line. With regard to the H9 cell line similar results were obtained at 6 h from infection. However, at 24 and 48 h the concentrations of saturated FAs returned to preinfection levels while the concentrations of unsaturated FAs dropped to levels even lower than those obtained at zero time. No significant changes in FA composition were found with PBLs.

Effect of a non-viral fraction of acquired immunodeficiency syndrome plasma on the vulnerability of lymphocytes to cortisol.

We have observed previously that the rate of cortisol catabolism by lymphocytes (CCL) was indicative of the vulnerability of these cells to cortisol. We attempted to ascertain whether cortisol-sensitive lymphocytes (e.g. thymocytes) metabolize cortisol at a different rate from cortisol-resistant cells and whether lymphocytes in which cortisol catabolism is inhibited become cortisol sensitive. The work was facilitated by the observation that an ethanol extract plasma from patients with acquired immunodeficiency syndrome (AIDS) and AIDS-related complex (ARC) had the capacity to inhibit CCL. The capacity of thymocytes to metabolize cortisol was found to be 11 times lower than that of peripheral lymphocytes. Inhibition of CCL with an ethanol extract of plasma from AIDS/ARC patients made the cells vulnerable to cortisol, causing them to die at a rate seven times greater than that of control samples. It is suggested that these findings may have important implications with regard to the nature of lymphocyte depletion in AIDS/ARC patients or in people at risk of developing the syndrome.

The effect of fatty acids on the vulnerability of lymphocytes to cortisol.

We have shown previously that cortisol-sensitive lymphocytes (thymocytes) have a much lower capacity than cortisol-resistant cells to catabolize cortisol and that linoleic acid inhibits the catabolism of cortisol by lymphocytes and modulates the sensitivity of lymphocytes to cortisol. In the present study, we attempted to see whether other fatty acids are inhibitory and if inhibition of cortisol catabolism by lymphocytes indicates a change in resistance of the cells to cortisol. Measuring the effect of fatty acids on cortisol catabolism by lymphocytes indicated that the polyunsaturated fatty acids, linoleate, arachidonate, and eicosapentaenoic, inhibit cortisol catabolism by lymphocytes. Using prostaglandin PGE2 and indomethacin as a blocker of prostaglandin formation, we observed that the effect of the polyunsaturated fatty acids was not due to the formation of prostaglandins. Examining the effect of fatty acids on the vulnerability of lymphocytes to cortisol, we noted that saturated fatty acids had no significant effect, whereas the aforementioned polyunsaturated fatty acids make lymphocytes more sensitive to cortisol.

The effect of butylated hydroxytoluene, with and without cortisol, on stimulated lymphocytes.

In the present work we undertook to ascertain whether butylated hydroxytoluene (BHT), which is used in food as an antioxidant, is capable of either inhibiting human lymphocyte stimulation or acting synergistically with cortisol and prednisolone to the same end. BHT cytotoxicity was observed at concentrations higher than 100 micrograms/ml. In the concentration range of 0.0 to 60.0 micrograms/mL, BHT showed no effect on the uptake of 3H-thymidine by PHA stimulated lymphocytes. However, at 50 micrograms/mL BHT suppressed mixed lymphocyte reaction (MLR). A synergistic effect with regard to suppression of PHA stimulated lymphocytes was observed when the cells were incubated with BHT in the presence of either cortisol or prednisolone.

The effect of linoleic acid on the sensitivity of human lymphocytes to cortisol and their capacity to catabolize the steroid.

We have shown previously that cortisol-sensitive lymphocytes (thymocytes) have a much lower capacity than cortisol-resistant cells to catabolize cortisol. In the present study, we attempt to demonstrate that inhibition of cortisol catabolism may make cortisol-resistant lymphocytes vulnerable to the steroid. Linoleic acid, which has the capacity to inhibit the catabolism of cortisol by lymphocytes, was used for this purpose. By using various concentrations of linoleic acid (20-60 micrograms/mL) we showed an inverse linear relationship between linoleic acid concentration and the rate of cortisol catabolism by lymphocytes. During this experiment which took 17 h the viability of cells did not change significantly (minimum viability 95%), even at the highest concentration of linoleic acid. Keeping the metabolism of cortisol at a level of 40% of that obtained by the control, by adding linoleic acid to lymphocyte cultures (50 micrograms/mL) and measuring the viability of the cells for a period of 3 days in the presence or absence of cortisol, we were able to show a rise in the death rate of the cells which started after 24 h of incubation owing to the presence of the steroid.

Effect of ethanol extract of cancer patients' serum on the vulnerability of lymphocytes to cortisol.

We have shown previously that cortisol-sensitive lymphocytes (thymocytes) have a much lower capacity than cortisol-resistant cells to catabolize cortisol. We have also shown that sera of cancer patients (CPS) possess ethanol-extractable substance(s) which can inhibit the catabolism of cortisol by lymphocytes (CCL). Recently, we noted that unsaturated fatty acids can both inhibit CCL and modulate the sensitivity of lymphocytes to cortisol. In the present study, we attempt to identify the compounds responsible for CCL inhibition and to demonstrate that inhibition of CCL may make cortisol-resistant lymphocytes vulnerable to the steroid. The enzymes DNase, RNase, pronase and lipase were added to ethanol extracts of serum as a first step in our efforts to identify the nature of the inhibitors of CCL. Only lipase had an effect on the inhibition. In fact, lipase enhanced the inhibition of CCL. This finding correlates with our previous observations that unsaturated fatty acids are potent inhibitors of CCL. Examining the effect of ethanol extracts of CPS and normal serum on the vulnerability of lymphocytes to cortisol, we noted that ethanol extracts of normal serum had no significant killing effect, whereas an ethanol extract of CPS makes lymphocytes more sensitive to cortisol. Since the adsorbance of free fatty acids of CPS by defatted albumin reduced but did not eliminate the capacity of the serum to inhibit CCL, we assume that other compounds besides free fatty acids might also be involved in CCL inhibition and modulation of the sensitivity of lymphocytes to cortisol.

Albumin and the unique pattern of inhibitors of cortisol catabolism by lymphocytes in serum of cancer patients.

We have shown previously that sera of cancer patients (CPS) possess ethanol-extractable substances which can inhibit the catabolism of cortisol by lymphocytes (CCL). In the present study an attempt was made to purify the inhibitory material by gel filtration. Chromatography of normal serum and CPS on a Sephadex G-10 column showed one peak of CCL inhibition with control serum and two peaks with CPS. The one peak which was common to both sera appeared with the void volume and was identified as albumin. The second peak which was present with CPS only, appeared at a molecular weight range of 300-350 daltons. We postulate that CPS may contain a relatively high concentrations of small molecules which are not bound to proteins and which might modulate the normal function of the immune system.

Cortisol catabolism by lymphocytes of patients with chronic lymphocytic leukemia.

A low rate of catabolism of cortisol by lymphocytes correlates with high sensitivity of the cells to the steroid and causes them to die at a greater rate than control samples. Since lymphocytes of patients with chronic lymphocytic leukemia respond to treatment with glucocorticosteroids and are cortisol sensitive, we attempted to see whether their capability to catabolize cortisol differs from that of normal lymphocytes. No difference was found between the two groups of cells with regard to the pattern of cortisol metabolites. However, the lymphocytes of the chronic lymphocytic leukemia groups showed a total cortisol catabolism per cell that was significantly lower than that of the control group. Patients with low lymphocyte count in peripheral blood showed a relatively higher cortisol metabolism by lymphocytes per cell than those with high counts.

Cortisol catabolism by lymphocytes of patients with systemic lupus erythematosus and rheumatoid arthritis.

We have shown that low cortisol catabolism by lymphocytes correlates with a high sensitivity of the cells to the steroid. In the present study, we aimed to assess whether high resistance to corticosteroid treatment correlates with a high rate of cortisol catabolism by lymphocytes. Since patients with systemic lupus erythematosus (SLE) usually require high doses of corticosteroids, while patients with rheumatoid arthritis (RA) respond to relatively low doses of steroids, we compared the capability of lymphocytes of patients with SLE and RA to catabolize cortisol. The rate of cortisol catabolism obtained with the RA group was not significantly different from that obtained with the control group. The catabolism of cortisol by lymphocytes of the SLE group was significantly higher than both the control group (p less than 0.05) and the RA group (p less than 0.01). A significant correlation was demonstrated between the SLE disease activity index and rates of cortisol catabolism attained by lymphocytes of SLE patients (p less than 0.001).