Suppression of B-lymphocyte function by T-lymphocytes in patients with advanced lung cancer.

The ability of B-lymphocytes to produce immunoglobulins in response to pokeweek mitogen stimulation was studied in 21 untreated stage III lung cancer patients by culture of their mononuclear cells in vitro. The number of immunoglobulin-producing cells was significantly lower in 20 of the 21 patients when compared to responses shown by normal control subjects. In contrast, the proliferative responses of many of the patients were within the normal range. When the T-lymphocytes of these patients were irradiated with 1,250 rad to eliminate the suppressor T-cell activity and then cultured with autologous B-cells, the number of immunoglobulin-producing cells was enhanced to the normal range in 7 of the 18 patients. These results indicate that B-cell function is impaired in most patients with advanced lung cancer. They also suggest that, in addition to suppression by radiosensitive suppressor T-cells, other mechanisms are involved in the observed B-cell functional abnormality.

Membrane components in the red cells of patients with sickle cell anemia. Relationship to cell aging and to irreversibility of sickling.

Cholesterol, phospholipid and sialic acid were measured in red cells from patients with sickle cell anemia to determine whether the cells had abnormal concentrations of these components and whether the amounts of these compounds differed in irreversibly sickled cells as compared to non-irreversibly sickled cells. Sickle cells had significantly higher levels of both lipids than similar populations of normal cells, however, comparisons to populations of young control cells showed that the differences were generally not significant. Sialic acid levels in sickle cells were not significantly different from normal cells. Irreversibly sickled cells had lower lipid and sialic acid concentrations than those not irreversibly sickled, but the differences were either not significant or did not occur when compared to young control cells. The studies show that the increased lipid concentrations in the membrane of sickle cells are not abnormal but are related to cell age and that the decrease in membrane components in irreversibly sickled cells is no greater than would be predicted for similarly aged populations of cells.

Reduced water exchange in sickle cell anemia red cells: a membrane abnormality.

We have measured the diffusional water permeability of sickle cell anemia red blood cells under isotonic conditions using pulsed nuclear magnetic resonance (NMR) techniques. We have found that the equilibrium diffusional permeability for sickle cells is about 1.61.10(-3) cm/s, or about 60% of the value measured for normal cells. This abnormality is not related to the heterogeneity generally found in cell populations in sickle red cells with different mean corpuscular hemoglobin concentrations. We speculate that the abnormality of water exchange under isotonic conditions in sickle cells reflects an alteration of membrane proteins responsible for water exchange, possibly caused by oxidation of Band 3 proteins.

Real-time X-ray diffraction study at different scan rates of phase transitions for dipalmitoylphosphatidylcholine in KSCN.

Multibilayer arrays of dipalmitoylphosphatidylcholine (DPPC) in 1 M KSCN were characterized using real-time X-ray diffraction and differential scanning calorimetry. A phase transition sequence was observed as a function of increasing temperature which involved changes from the interdigitated subgel (Lc(inter)) to interdigitated gel (L beta(inter)) to disordered (L alpha) bilayer states. The phase transition mechanisms were unambiguously determined by comparison of results from fast and slow scans. The Lc(inter)-->L beta(inter) phase transition was shown to involve a continuous change in acyl chain spacing between the rectangular subgel acyl chain unit cell into an hexagonal gel acyl chain unit cell. The mechanism is similar to that for subgel to gel state transitions involving non-interdigitated DPPC bilayers.

Sterols stabilize the ripple phase structure in dihexadecylphosphatidylcholine.

The presence of various sterols in mixtures with dihexadecylphosphatidylcholine (DHPC) was studied using static X-ray diffraction of temperature equilibrated samples, and real-time X-ray diffraction of samples undergoing temperature scans. It was found that these sterols eliminate the interdigitation of the alkyl chains in the DHPC sub-gel and gel-state bilayers while stabilizing the ripple gel-state at the expense of the gel-state bilayer phase. The ripple-ripple phase transition previously observed for dipalmitoylphosphatidylcholine in the presence of low molar concentrations of sterols (Wolfe et al. (1992) Phys. Rev. Lett. 68, 1085-1088) was also observed for similar DHPC-sterol mixtures. In addition, we show the first evidence that the presence of 5 alpha-cholestane-3 beta,5,6 beta-triol will cause the lipid mixtures to continue to adopt a ripple mesophase structure even after the DHPC alkyl chain becomes disordered.

The effects of cholesterol oxidation products in sickle and normal red blood cell membranes.

The oxysterol content in normal and sickle red blood cell (RBC) membranes was assessed using thin-layer chromatography and capillary gas chromatography/mass spectrometry. Several more oxysterols were present in sickle RBCs compared to normal RBCs. Sickle RBC membranes had a higher concentration of 5 alpha,6 alpha-epoxycholesterol, 5 alpha-cholestane-3 beta,5,6 beta-triol, 7-ketocholesterol and 19-hydroxycholesterol than normal RBC membranes. The increased oxysterols in sickle RBC may be an effect of the increased oxidative stress which occurs in sickle RBC membranes. Physical characteristics of normal and sickle RBC membrane ghosts with and without inserted oxysterols were examined by Fourier transform infrared spectroscopy. The data are consistent with a greater sterol content in sickle cells compared to normal RBC membranes, and a possible oxysterol-cholesterol synergism.

Asplenia and abnormal neutrophil kinetics in chronic idiopathic neutrophilia.

Chronic idiopathic neutrophilia is a rarely recognized finding in otherwise healthy subjects. Described here is a case, not previously reported; in which the patient also had congenital asplenia. Studies carried out to determine the mechanism for the neutrophilia are also described. The results show that the patient's neutrophilia was associated with an enlarged circulating pool of neutrophils, increased production and utilization of neutrophils and a short blood granulocyte survival. The findings differ completely from those observed in patients with chronic idiopathic neutrophilia and intact spleens. The results suggest that (1) the mechanism for neutrophilia in patients with chronic idiopathic neutrophilia varies and depends upon the presence or absence of the spleen, and (2) the spleen has a role in the control of neutrophil production, distribution and utilization.

Subpopulation of human helper and suppressor T lymphocytes.

Mitogen driven differentiation of normal human mononuclear cells is a well-established model for the study of antibody synthesis in man. In certain rare individuals who are clinically normal, unfractionated mononuclear cells or a mixture of purified B plus T lymphocytes differentiate into immunoglobulin producing cells in response to purified protein derivative of tuberculin (PPD) but not in response to pokeweed mitogen (PWM). To evaluate this observation we have irradiated T cells from such individuals to eliminate naturally occurring suppressor T cell activity and then added the irradiated T cells back to autologous B cells before culture. The B cells then responded to PWM. The original PPD responses of cells from these individuals were now significantly reduced. Although, there was no difference between PWM nonresponders and responders in the number of OKT-8 positive cells, elimination of OKT-8 positive cells in the PWM nonresponders with OKT-8 monoclonal antibody and complement resulted in a significantly increased response to PWM. This study indicates that there are suppressor T cells which specifically inhibit B cell response to PWM without affecting the PPD response. These results also show that the helper T cells involved in the PWM response are radioresistant and those involved in the PPD response are radiosensitive.

B-cell changes occur in patients with sickle cell anemia.

B- and T-lymphocytes were studied in patients with sickle cell anemia. Samples were analyzed during asymptomatic periods and during vasocclusive pain crises to assess a possible relationship to these periods. Assays of B-cell function and quantitation of B- and T-lymphocytes and their subsets were carried out. During crises, six of the eight patients showed a significant decrease in the number of immunoglobulin-producing cells, together with normal or enhanced blastogenic responses. The total number of T-lymphocytes was normal when measured by the monoclonal antibody OKT3 in contrast to the significantly lower level observed in some patients, as determined by the E-rosette technic. There were no significant changes in the percentage of either helper or suppressor T-cells or in the percentage of B-cells. The results show that changes in in vitro B-cell function occur during vasocclusive pain crises in patients with sickle cell anemia. These may be clinically important in these patients.

Red blood cell morphology in sickle cell anemia as determined by image processing analysis: the relationship to painful crises.

Red blood cell morphology was studied in the peripheral blood of adults with sickle cell anemia to determine if changes occur during painful crises. Image processing of the cells with an automated system of red blood cell analysis was used. Four groups of cells were observed: normocytes, macrocytes, target cells, and cells with the shape of irreversibly sickled cells. During asymptomatic periods, the percentages of these cells differed in each individual but were typical for that individual and generally were stable. During crises, macrocytosis occurred and the concentration of irreversibly sickled cells showed greater fluctuation. The macrocytosis most likely reflected a marrow response to increased hemolysis and demonstrated that the increased red blood cell destruction observed during pain crises may be more extensive than previously considered. Changes in the concentration of irreversibly sickled cells during crises were not consistent and could not be used as an indicator of a crisis. Image processing with automated red blood cell analysis allows for accurate assessment of all the morphologic groups of red blood cells in patients with sickle cell anemia and compares well with standard methods for measuring the concentration of irreversibly sickled cells.

Assessment of the anemia of chronic disease by digital image processing of erythrocytes.

Eighteen patients with the anemia of chronic disease were studied to determine the usefulness and accuracy of image processing analysis of erythrocytes for diagnosis. Diagnostic tests used for comparison included estimates of serum ferritin, serum iron and iron-binding capacity, stainable marrow iron, and erythrocyte morphology as determined by standard methods. The findings show that the analyses obtained by image processing were diagnostic for 83% of the patients with chronic disease. Serum ferritin levels were supportive of diagnosis for 33% of the patients, and serum iron levels were useful for approximately 25% of the patients. Cell indices and marrow iron were of limited value. The study demonstrates that quantitative information obtained by digital image processing of erythrocytes can be very useful for the diagnosis of the anemia of chronic disease.

An evaluation of cellularity in various types of bone marrow specimens.

Comparisons of marrow cellularity determinations in needle biopsy sections, aspirated smears of marrow particles, and aspirated clot sections were made using the point-counting and standard visual methods of measurement. Good correlations of cellularity between biopsies and smears were obtained with the point-counting method. This occurred with either single- or multiple-observer measurements, and differed from the wide observer-to-observer variation obtained in the other comparisons. Although the optimal method for cellularity measurements is point counting by biopsy sections, the findings indicate that aspirated smears of marrow particles can be a suitable substitute for needle biopsy sections when point counting is used. This would be particularly useful in clinical settings necessitating frequent cellularity assessments. The results also show that the standard visual method of cellularity measurement is satisfactory when specimens are evaluated by one observer. The point-counting method of measurement, although the most accurate, is not as reproducible among observers as previously considered.

Coexistent double gammopathy, myeloproliferative disorder, and malignant lymphoma.

The authors report a patient with coexistent double gammopathy, a Philadelphia chromosome-negative, bcr rearrangement-negative myeloproliferative disease resembling chronic myelocytic leukemia and a malignant lymphoma of B-cell origin. The double gammopathy consisted of IgM (kappa) and IgG (kappa). Peripheral blood, spleen, and marrow lymphocytes had primarily an IgG (kappa) isotype, whereas lymph node lymphocytes had predominantly an IgM (kappa) surface isotype. Increased numbers of marrow lymphocytes stained doubly for both IgM (kappa) and IgG (kappa). The results suggest that doubly isotypic as well as single isotypic lymphocytes contributed to the double gammopathy. Organ localization differed for lymphocytes with different antibody isotypes. This cluster of findings has not been described previously.

Chronic anemia, wound healing, and red cell 2,3-diphosphoglycerate.

Relationships between various types of chronic anemia, wound healing, and red cell 2,3-diphosphoglycerate (2,3 DPG) were examined in rabbits. Wound tensile strength and energy absorption were not affected by chronic iron-deficiency anemia, the chronic hemolytic anemia caused by intravenous water infusion nor by chronic hemolytic anemia caused by intravenous water infusion nor by chronic phenylhydrazine-induced anemia. Red cell 2,3 DPG levels were increased in the anemia of iron deficiency and were normal in the rabbits with chronic phenylhydrazine-induced anemia at the time of wound excision but were low following phynylhydrazine injection. The results show that chronic anemia per se does not affect the tensile strength and energy adsorption of wound healing. The findings suggest that the wound healing process may differ in certain types of anemia.

Effects of fibrinogens on phase transitions in lipid model membrane systems.

An abnormal fibrinogen that caused aggregation of red blood cells (RBC) in a patient with gangrene was examined by real-time X-ray diffraction to determine its effects on dipalmitoylphosphatidylcholine (DPPC) and 1-palmitoyl-2-oleoylphosphatidylethanolamine (POPE) phase transitions. Similar studies were done with normal fibrinogen and results were compared. Both types of fibrinogen slightly increased the L alpha-->HII phase transition temperature and the HII phase parameters for POPE, while neither fibrinogen significantly affected the order-disordered acyl chain transitions in the lipid bilayer phase. However, fibrinogen differentially influenced the bilayer unit cell parameter of the gel and disordered bilayer and the gel state ripple phase. These results can be interpreted as indicating that fibrinogen has little effect on the balance of gel and disordered acyl chains in the lipid bilayer, but may influence membrane functions dependent on non-bilayer phases.

Inhibition of cytolytic T lymphocyte activity by oxysterols.

The objective of this study was to investigate the effects of oxysterols (OS), namely 5 alpha-hydroxy-6-ketocholestanol, 6-ketocholestanol and 25-hydroxycholesterol, on specific cell-mediated cytotoxicity by C57BL/6 spleen cells against P815-X2 (a DBA/2 mastocytoma) target cells. Cytolytic T lymphocytes (CTL) were generated by intraperitoneally injecting C57BL/6 mice with P815-X2 tumor cells 10 d prior to the cytotoxicity experiments. Preincubation of CTL with 10(-5) M 5 alpha-hydroxy-6-ketocholestanol and 6-ketocholestanol for 45 min in lipoprotein-depleted medium resulted in an inhibition of cytolytic activity (73 and 43%, respectively) as measured by 4-h 51Cr release. At a concentration of 5 x 10(-6) M, 5 alpha-hydroxy-6-ketocholestanol inhibited CTL activity by 65%, whereas 6-ketocholestanol did not elicit any inhibition. By contrast, 25-hydroxycholesterol did not inhibit CTL at either concentration, although it is known to be a potent inhibitor of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase, the rate-limiting enzyme in the cholesterol biosynthetic pathway. When CTL were preincubated with OS in lipoprotein-replete medium, there was no inhibition of CTL activity at the respective concentrations. The results suggest that the inhibition of CTL activity upon short-term incubation with OS is not due to the inhibition of cholesterol synthesis, but may be due to the insertion of OS into the plasma membrane to replace cholesterol and alteration of membrane physical properties.