Stimulation of lymphocytes from subacute sclerosing panencephalitis patients by defined measles virus antigens

The lymphoproliferative response of human peripheral blood mononuclear cells to different measles virus antigen preparations was studied with lymphocytes from 38 measlesseropositive healthy donors and 4 subacute sclerosing panencephalitis patients. The response was very weak or absent in all of the controls and in three of the subacute sclerosing panencephalitis patients. The fourth subacute sclerosing panencephalitis patient had fluctuating levels of lymphocyte stimulation by measles antigens. The response was very strong for several months and during this time the parameters of the test system were characterized. It was discovered that a membrane preparation of measles-infected cells caused stimulation equal to that of highly purified virions. Purified measles ribonucleoprotein also induced specific stimulation, although lower than that seen with other types of measles antigens. Results of experiments on stimulation kinetics and antigen dose responses were compatible with antigen-specific stimulation. Enriched T cells were more vigorously stimulated than unfractionated peripheral blood mononuclear cells suggesting that this transformation test is specific for T cells.     

Skin-window study on the migration of leukocytes of newborns and infants

Migration of leukocytes of newborns and of infants up to the age of 6 months was studied using the in vivo skin-window technique according to Rebuck. Using the non-specific stimulation (abrasion of the skin only) a slight age-dependent physiological increase of migration of cells was observed within the observation period; after a strong local irritation with diphtheria-tetanus-pertussis vaccine (Alditepera) there was a vigorous migratory response of cells in the skin lesion. Both quantitatively and qualitatively, the migration pattern (i.e. shift from PMN leukocyte to mononuclear cells in the exudate within a 1 d period after abrasion) was not influenced by immunization of infants with Alditepera, suggesting thus the nonspecific nature of this cellular response. The "normal" values of the chemotactic response of leukocytes of newborns and infants are given as a basis for evaluation of defects of this functional activity of leukocytes.     

Effect of somatotropic hormone on autoimmune responses induced in mice of different genotypes by syngenous heated erythrocytes]

A single administration of 1 X 10(9) heated erythrocytes to C57BL and BALB/c mice caused on the 13th day the appearance of antierythrocytic autoantibodies, an increase in the weight of the lymphoid organs, and lymphoreticular hyperplasia. These changes were more pronounced in BALB/c mice. During the development of autoimmune reactions the changes in the number of E- and EAC-rosette-forming cells in the thymus and spleen and in the immune response to the sheep erythrocyte immunization and E. coli endotoxin were revealed; distinct strain differences were observed. Daily somatotropic hormone administration (5 mg/kg of body weight) for 10 days decreased the degree of the autoimmune reactions development in mice of both strains. Its action was more expressed in BALB/c mice.     

Human newborn natural killer cell responses to activation by monoclonal antibodies. Effect of culture with herpes simplex virus

Deficient responses by NK cells may contribute to the susceptibility of human newborns to severe HSV infections. Furthermore, HSV disseminates widely during neonatal infection and may also infect and interfere with the function of blood mononuclear cells (MNC). To determine whether the function of newborn NK cells would be affected by contact with HSV, and whether newborn NK cells might be permissive for HSV replication, newborn MNC were cultured with HSV in vitro for 3 days. Before culture, the intracellular calcium mobilization in newborn NK cells induced by mAb to CD2 and CD16 did not differ from that of adult NK cells. This result argues against immaturity of an early NK cell activation event in human newborns. After culture with HSV the Ca2+ flux response was unaffected by lysis of K562 targets by newborn NK cells and NK-dependent suppression of HSV replication in fibroblasts were preserved or increased. Incorporation of thymidine by NKH-1 cells following stimulation with PHA and IL-2 was not suppressed. NK cells recovered from these cultures did not contain infectious HSV and synthesis of HSV-specific proteins was not detected by immunoprecipitation although HSV genome was detected by DNA hybridization. Our results extend the in vitro model stimulation systems in which newborn NK cells respond positively to include triggering through the CD2 Ag, cross-linking of cell surface CD 16 Ag and response to a pathogen, HSV.     

Manifestation of radiation injury of human lymphocytes using PHA mitogenic stimulation in different culture systems

The proliferative response of human lymphocytes to PHA in vitro is affected by X-irradiation. Dose-related changes of mitogenic stimulation of irradiated lymphocytes were compared in two culture systems--cultivation of separated lymphocytes and cultivation of whole blood. In whole blood cultures, the proliferative activity of stimulated lymphocytes was markedly and reproducibly depressed by irradiation. The values of mitogenic response within a dose range from 0 to 2.5 Gy could be fitted with high correlation by an exponential curve. In a modified test where the mitogenic stimulus was given after 24 h delay, depression of the response was even more pronounced. Radiosensitivity of human lymphocytes as determined by means of mitogenic stimulation in whole blood cultures appears to be a characteristic individual feature. The mean D37 value of the radiation-induced depression of mitogenic response in a group of 20 healthy donors was 2.5 Gy in the standard test and 2.0 Gy in the test with a delayed mitogenic stimulus. In contrast, the data obtained from separated lymphocyte cultures were characterized by a high degree of the test-to-test variability and by much lower radiosensitivity. The possible mechanisms of these distinctive manifestations of the same primary radiation injury are discussed.     

A monokine stimulates prostaglandin-E2 production by human amnion

The studies reported in this communication were designed to test the hypothesis that products of mononuclear cells are capable of stimulating prostaglandin E2 production by human amnion. Conditioned media obtained from peripheral blood mononuclear cells were incubated with amnion cells in primary culture. A dose dependent increase in PGE2 biosynthesis was observed in response to increasing amounts of the conditioned media. These observations suggest that mononuclear cells produce a factor(s) capable of stimulating prostaglandin production by amnion cells. The signal responsible for the increased biosynthesis of prostaglandins by human amnion associated with parturition in the setting of intraamniotic infection may be of host origin.     

Lymphocyte reactivity in pregnant women and newborn infants.

The mitotic response to phytohaemagglutinin (PHA) was determined in lymphocytes of mothers and their newborn infants obtained at delivery and seven days later by measuring the rate of 125 I-idoxuridine uptake into DNA in lymphocytes cultured in their own plasma and after washing and resuspension in fetal bovine serum. There was no difference in the unstimulated counts of maternal lymphocytes taken at delivery, whether unwashed or washed, compared with those from nonpregnant controls. With PHA stimulation the mitotic response of the maternal lymphocytes cultured in their own plasma was reduced compared with that of the control lymphocytes but washed maternal cells showed a similar response to the controls. These findings suggest that the reduced lymphocyte mitotic response to PHA in pregnancy is due to a plasma inhibitory factor This inhibition was not evident in maternal blood taken seven days after delivery. DNA synthesis in unstimulated cultures from newborn infants at birth and seven days after birth was greater than that in adult control cultures. With PHA stimulation the mitotic response of cord-blood lymphocytes cultured in their own plasma paralleled that of control lymphocytes but washed newborn cells showed a greater response. Thus plasma suppression similar to that observed in the mother seems also to affect infants at birth. This inhibition was not demonstrable in blood taken from infants of 7 days.     

T-cell immune response to human herpesvirus-6 in healthy adults.

The proliferative response of peripheral blood mononuclear cells (PBMC) from healthy adults to human herpesvirus-6 (HHV-6) was examined. It was found that PBMC from 13 of 14 HHV-6-seropositive adults apparently proliferated in response to stimulation with HHV-6 antigen in contrast to the lack of response of cord blood mononuclear cells. In order to confirm the presence of HHV-6-specific memory T cells in the peripheral blood of healthy adults, we established HHV-6-specific T-cell clones from an HHV-6-seropositive individual. CD4+ T-cell clones generated from HHV-6-stimulated PBMC were found to proliferate upon stimulation with HHV-6 in the presence of autologous antigen-presenting cells, but not in response to herpes simplex virus type 1 antigen or mock-infected control antigen. These results indicate that a T-cell immune response against HHV-6 infection is generally present in healthy adult populations.     

Peripheral blood mononuclear cell production of TNF-alpha in response to North American ginseng stimulation

North American ginseng (Panax quinquefolium) root extract (NAGE) with known ginsenosides composition was examined for its affinity to stimulate human tumour necrosis factor alpha (TNF-alpha) production in human peripheral blood mononuclear cells. Case studies were conducted in three donors, one that was diagnosed with an atopic allergy and two that were normal, healthy subjects. Cultured mononuclear cells were incubated with varying concentrations of NAGE for up to 72 h and culture media were tested for TNF-alpha concentration. Direct stimulation of mononuclear cell TNF-alpha production in vitro by NAGE occurred as early as 6 h with 200 microg NAGE/mL. The stimulation of TNF-alpha production was confirmed by TNF-alpha mRNA gene expression. These interesting results show the immunostimulating activity of NAGE components in reference to TNF-alpha production. This observation requires further investigation with more subjects to determine the affinity of ginseng in stimulating the human immune system. Moreover, the method of evaluating this response is very useful for standardizing ginseng extracts to a known bioactivity.     

The Influence of Regular Physical Activity on the Cell-Mediated Immunity in Pigs

The influence of moderate regular physical activity on the cell-mediated immunity was studied in growing pigs. Ten animals were subjected to physical training on a large animal treadmill, and 10 were kept in their pens throughout a 12-week experimental period. Regardless of whether the pigs underwent training or not, a whole blood lymphocyte stimulation test performed at 3 stages of the experiment revealed an equal ability of the cells to respond to stimulation induced by pokeweed mitogen and phytohaemaglutinin. The influence of serum from the pigs of the trained and untrained groups was studied in a stimulation test with purified mononuclear cells obtained from 2 healthy control pigs. The results indicated that no additional serum factors released by the physical training altered the blastogenic response of these lymphocytes. It is concluded that moderate exercise should not be regarded as a stressor which alters the cellular immunity in pigs.     

Stimulation of production of prostaglandin E in gingival cells exposed to products of human blood mononuclear cells.

1. Supernatant media from cultures of unstimulated human peripheral blood mononuclear cells contained one or more factors that increased by several hundred-fold the production of prostaglandin E by fibroblast-like cells derived from both inflamed and normal human gingival tissue. 2. This stimulation occurred in a dose-dependent manner and was completely inhibited by 14 microM-indomethacin. 3. Responsiveness to the factor declined as the age of the cell culture increased. 4. An increase in prostaglandin E production was first observed after a 2h exposure to the mononuclear cell factor(s) and could be prevented by cycloheximide. 5. Brief exposure (0.5 and 1.0 h) to mononuclear cell factor did not increase prostaglandin E production by the cells in a subsequent 72 h incubation in the absence of mononuclear cell factor. 6. Addition of arachidonate (10 microM and 15 microM) further enhanced stimulation of prostaglandin E production in response to mononuclear cell factor. 7. The stimulatory activity was resistant to digestion by trypsin, but was heat-labile, so that only 17% remained after treatment at 56 degrees C for 30 min.     

Postnatal development of E-4031-sensitive potassium current in rat carotid chemoreceptor cells.

The O2 sensitivity of dissociated type I cells from rat carotid body increases with age until approximately 14-16 days. Hypoxia-induced depolarization appears to be mediated by an O2-sensitive K+ current, but other K+ currents may modulate depolarization. We hypothesized that membrane potential may be stabilized in newborn type I cells by human ether-a-go-go-related gene (HERG)-like K+ currents that inhibit hypoxia-induced depolarization and that a decrease in this current with age could underlie, in part, the developmental increase in type I cell depolarization response to hypoxia. In dissociated type I cells from 0- to 1- and 11- to 16-day-old rats, using perforated patch-clamp and 70 mM K+ extracellular solution, we measured repolarization-induced inward K+ tail currents in the absence and presence of E-4031, a specific HERG channel blocker. This allowed isolation of the E-4031-sensitive HERG-like current. E-4031-sensitive peak currents in type I cells from 0- to- 1-day-old rats were 2.5-fold larger than in cells from 11- to 16-day-old rats. E-4031-sensitive current density in newborn type I cells was twofold greater than in cells from 11- to 16-day-old rats. Under current clamp conditions, E-4031 enhanced hypoxia-induced depolarization in type I cells from 0- to- 1-day-old but not 11- to 16-day-old rats. With use of fura 2 to measure intracellular Ca2+, E-4031 increased the cytosolic Ca2+ concentration response to anoxia in cells from 0- to- 1-day-old but not cells from 11- to 16-day-old rats. E-4031-sensitive K+ currents are present in newborn carotid body type I cells and decline with age. These findings are consistent with a role for E-4031-sensitive K+ current, and possibly HERG-like K+ currents, in the type I cell hypoxia response maturation.     

Increased blood spermine levels decrease the cytotoxic activity of lymphokine-activated killer cells: a novel mechanism of cancer evasion

Increased blood polyamine levels, often observed in cancer patients, have negative impacts on patient prognosis and are associated with tumor progression. The purpose of our study was to examine the effects of polyamines on cellular immune function. Peripheral blood mononuclear cells (PBMCs) from healthy volunteers were cultured with the human natural polyamines spermine, spermidine, or putrescine, and the effects on immune cell function were examined. The correlation between post-operative changes in blood polyamine levels and lymphokine-activated killer (LAK) activity was also examined in cancer patients. Spermine decreased the adhesion of non-stimulated PBMCs to tissue culture plastic in a dose- and a time-dependent manner without affecting cell viability or activity. This decrease in adhesion capacity was accompanied by a decrease in the number of CD11a bright-positive and CD56 bright-positive cells. Upon stimulation with interleukin 2 to activate LAK cytotoxicity, PBMCs cultured overnight with 100 or 500 μM spermine showed decreased cytotoxic activity against Daudi cells (91.5 ± 1.7 and 84.9 ± 3.0%, respectively (n = 6) compared to PBMC cultured without polyamines). In a group of 25 cancer patients, changes in blood spermine levels after surgery were negatively correlated with changes in LAK cytotoxicity after surgery (r = −0.510, P = 0.008: n = 25). Increased blood spermine levels may be an important factor in the suppression of anti-tumor immune cell function.     

Immunological analysis of human lymphocytes bearing different surface receptors]

Cell surface receptors on human lymphocytes being studied, essential differences were revealed in a relative content of E-, EA- and EAC-rosette-forming cells in peripheral blood and tonsils. In tonsils, part of lymphocytes carrying receptors for sheep erythrocytes have been shown to possess C'3-receptors for sheep erythrocytes. Apparently, C'-receptor,--being B-lymphocyte surface marker,--may also appear at definite stages of T-cell differentiation.     

Impaired local natural killer cell activity in human colorectal carcinomas

Summary The present study was undertaken to study natural killer (NK) cell activity in patients with colorectal cancer at peripheral and local levels. Mononuclear cells were isolated from uninvolved colorectal mucosa, tumor tissue and peripheral blood, and tested against the colon carcinoma cell line CaCo-2 and the erythroleukemia cell line K-562. Peripheral blood NK cell activity from the patients showed similar levels compared with healthy controls, whereas, mononuclear cells of tumor tissue were found to have a significantly decreased NK cell activity compared to the normal intestinal mucosa (P<0.01). No relation was found between the NK cell activity and the advancement of the disease according to the Duke's stage. Interferon- (IFN-) stimulated the NK cell activity of the mononuclear cells from blood, mucosa and tumor. However, the increase of NK cell activity after IFN- stimulation was lower in the tumor compared to the mucosa (P<0.02). The lectin, phytohaemagglutinin, increased the cytotoxicity of mononuclear cells from blood, mucosa and tumor to a similar level. These results suggest that patients with colorectal tumors exhibit a normal NK cell activity in peripheral blood and intestinal mucosa; however, a diminished NK cell activity exists at the tumor level. Although mononuclear cells isolated from the tumor have a normal response to lectin stimulation they show hyporesponsiveness to IFN- stimulation with regard to their NK cell activity.     

Virologic and serologic studies on an outbreak of echovirus type 11 infection in a hospital maternity unit.

An outbreak of echovirus type 11 (E-11) infection occurred among newborn babies in a hospital maternity unit in the summer of 1971. The results of studies are as follows: 1) Forty-one of 188 infants developed febrile illness with stomatitis during one and a half months from July to September. E-11 was isolated from stool specimens of 14 infants and two throat swabs. Antibody response to the virus was shown in all the 19 cases examined. Some of their mothers were suffering from subclinical infection. 2) The isolates were identified as a variant of E-11 which is not neutralized with antiserum against prototype E-11. Antiserum against the current virus neutralized both current and prototype viruses. 3) Sucrose gradient centrifugation of sera from infants revealed that the neutralizing antibody activity resided more predominantly in 19S than in 7S fractions. These antibodies reacted more specifically with the current strain than with the prototype Gregory strain.     

Competition of IL-1 and IL-1ra determines lymphocyte response to delayed stimulation with PHA.

BACKGROUND: Human peripheral blood mononuclear cells (PBMC) left in microcultures for 24h without mitogen do not respond to subsequent stimulation with PHA. They regain reactivity if the native culture medium is absorbed with other party lymphocytes or partially replaced with the medium from a PHA-stimulated culture. The observations suggest that, during the incubation, some inhibitory agent had accumulated in the culture medium. AIM: The study was performed to determine the nature of the observed phenomenon in respect of the possible role of monocytes and their products IL-1 and IL-1 receptor antagonist (IL-1ra), and to test for immunodiagnostic purposes the significance of quantifying the lymphocyte response to delayed stimulation with PHA in patients suffering from inflammatory prosesses. METHODS: Lymphocyte response to delayed stimulation with PHA, calculated as the lymphocyte-monokine interaction (LM) index, was determined in the microcultures of PBMC isolated from the blood of healthy donors or of patients with acute tonsilitis. The values of LM indices were compared with the ratios of IL-1ra/IL-1beta concentration estimated by enzyme-linked immunosorbent assay method in the culture supernatants. The influences of exogenous IL-1beta, IL-1ra, anti-IL1ra antibodies and antibiotic cefaclor on the monokine concentrations and on the values of LM index were tested. RESULTS AND CONCLUSIONS: The results show that the level of lymphocyte response to delayed stimulation with PHA (LM index) is inversely proportional to the ratio of IL-1ra/IL-1beta concentration in the culture. The low LM values at high IL-1ra/IL-1beta ratios in PBMC cultures from healthy donors, reversed proportions found in patients'' PBMC (acute tonsilitis), and the cefaclor-induced reduction of LM value with correlated increase of the IL-1ra/IL-1beta ratio suggest that the LM assay may prove to be useful for immunodiagnostic purposes.     

Loss of EGF binding and cation transport response during differentiation of mouse neuroblastoma cells

Mouse neuroblastoma cells (clone N1E-115) differentiate in culture upon withdrawal of serum growth factors and acquire the characteristics of neurons. We have shown tht exponentially growing N1E-115 cells possess functional epidermal growth factor (EGF) receptors but that the capacity for binding EGF and for stimulation of DNA synthesis is lost as the cells differentiate. Furthermore, in exponentially growing cells, EGF induces a rapid increase in amiloride-sensitive Na+ influx, followed by stimulation of the (Na+-K+)ATPase, indicating that activation of the Na+/H+ exchange mechanism in N1E-115 cells [1] may be induced by EGF. The ionic response is also lost during differentiation, but we have shown that the stimulation of both Na+ and K+ influx is directly proportional to the number of occupied receptors in all cells whether exponentially growing or differentiating, thus only indirectly dependent on the external EGF concentration. The linearity of the relationships indicates that there is no rate-limiting step between EGF binding and the ionic response. Our data would suggest that as neuroblastoma cells differentiate and acquire neuronal properties, their ability to respond to mitogens, both biologically and in the activation of cation transport processes, progressively decreases owing to the loss of the appropriate receptors.     

The effect of using T-activin in the therapy of the newborn with suppurative surgical infection

A total of 156 newborn infants with suppurative surgical infection (SSI) were observed; 73 of them had sepsis and 83 a severe localized process. In 47 patients with sepsis and 34 with localized infection, T-activin was included in complex therapy while the other infants formed the control group. It has been established that T-activin leads to an increase in the quantity of the active population of T-lymphocytes in the peripheral blood and to enhanced functional activity of T-lymphocytes in the newborn with SSI independent of generalization of the process. Bactericidal activity of circulating phagocytes is improved. The clinical course of SSI is less severe with more pronounced positive changes in the symptoms, hospital stay of the children is shortened, lethality is reduced. The effect of T-activin on the dynamic of the indices of the immune state is more marked in a septic process.     

Time-dependent loss of prostaglandin E release by adherent cells in response to stimulation by thyroid cells: Prevention of this loss by phytohemagglutinin

Preplating human adherent peripheral blood mononuclear cells (PBMC) for up to 24 hr results in a progressive decrease in their basal PGE release, and in the loss of their ability to increase PGE release during a subsequent 72-hr coculture period with allogeneic human thyroid cells. Phytohemagglutinin (PHA) present during a 24-hr adherent-cell preplating period prevents, in part, the loss of this PGE response to thyroid cells. These data indicate that adherent cells require continual stimulation by the thyroid cells or by PHA in order to maintain their ability to increase PGE secretion in response to thyroid cells.