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Immunologic tolerance to HGG in mice. I. Suppression of the HGG response in normal mice with spleen cells or a spleen cell lysate from tolerant mice.

Adoptive transfer of spleen cells or spleen cell lysates from mice tolerant to human-gamma-globulin (HGG) specifically suppressed the response of normal syngeneic recipients to HGG. The suppressive activity could be transferred for over 100 days after tolerance induction. The suppression induced by both spleen cells and spleen cell lysate was found to be specific as evidenced by a normal response to a challenge with turkey-gamma-globulin or goat erythrocytes. The activity of the suppressive lysate could be removed by passing the material through an HGG immunoadsorbent column but not by passing it through an anti-HGG column or a BSA column. These results indicated that the factor had antigen specificity and was probably not antigen-antibody complexes. That this suppression was not due to a shifting of the kinetics of the antibody response has also been demonstrated. The antigen-specific suppressor factor in the tolerant spleen cell lysates was a protein with a m.w. of approximately 45,000 daltons. The kinetics of the appearance of both suppressor cells and suppressor factor were consistent with a mechanism of active suppression functioning in the maintenance of tolerance to HGG.

Animals↗

Effects of chemicals on delayed matching behavior in pigeons. II: tolerance to the effects of diazepam and cross tolerance to phencyclidine.

Pigeons were trained under a 2-sec matching to sample schedule. Diazepam, phencyclidine and pentobarbital all decreased matching accuracy. Chronic administration of diazepam after the session produced no effect on matching accuracy on the next day; however, two weeks of postsession diazepam administration produced tolerance to the effects of presession diazepam administration and cross tolerance to phencyclidine, but not to pentobarbital. After 6 weeks without chronic diazepam administration, both the tolerance to the effects of diazepam on matching accuracy and the cross tolerance to phencyclidine had disappeared.

Animals↗

Delineation of tolerance-sensitive and tolerance-insensitive B cells in normal and immune defective mice.

B cells responsive to TNP on various thymus-independent carriers acquire immunocompetence at different times during ontogeny and elicit varying levels of responsiveness in immune-defective CBA/N mice. Such B cells have now been found to display differential tolerance susceptibilities in a hapten-specific in vitro tolerance assay. B lymphocytes from normal neonatal or immune-defective adult CBA/N mice responsive to TNP-lipopolysaccharide were shown to be hypersusceptible to tolerogenesis whereas B cell responses to a second TI antigen, TNP-Brucella abortus, were not rendered tolerant until high concentrations of tolerogen had been presented. Therefore, tolerance hypsersusceptibility is characteristic of a fraction of neonatal and immune-defective adult CBA/N antigen-reactive B lymphocytes.

Aging↗

Immunologic tolerance after oral administration of reovirus: requirement for two viral gene products for tolerance induction.

We have demonstrated that reovirus type 1, but not type 3, generates serotype-specific immunologic tolerance for DTH responses following oral administration of UV-inactivated virus. As shown by adoptive transfer experiments, the tolerance is secondary to the generation of viral-specific suppressor T cells that are present in spleen and mesenteric lymph nodes. No tolerance was generated when live virus was administered orally. Using recombinant viral clones, it was found that 2 viral gene products were required for tolerance induction; serotype specificity is a property of the viral hemagglutinin (the product of the S1 dsRNA segment). The inability of type 3 reovirus to induce suppression following oral administration is a property of the mu 1C polypeptide (the product of the M2 dsRNA genome segment).

Administration, Oral↗

The fate of allogeneic grafts of intact bone marrow in immunologically tolerant recipients and after abrogation of the tolerance.

In ossicles derived from grafts of compatible intact bone marrow in the subcapsular renal site the ectopic bone remains of donor provenance but the haemopoietic elements are partially replaced with tissue by host cells derived from stem cells migrating from the blood. Ossicles derived from incompatible intact bone marrow grafted in immunologically tolerant recipients are morphologically identical to those derived from compatible intact bone marrow. On abolition of the tolerance the allogeneic ectopic bone and donor microenvironment for haemopoiesis became manifest: haemopoietic marrow, even the host elements, was rapidly lost, bone disappearing rapidly if recently established, slowly if long established. Provided the state of immunological tolerance persists, the histoincompatible bone and microenvironment of the donor co-exist with marrow derived from the circulating haemopoietic stem cells of the tolerant recipient.

Animals↗

Mice tolerant to nitrous oxide are also tolerant to alcohol.

Mice exposed to 50% nitrous oxide for 3 weeks become tolerant to nitrous oxide. The nitrous oxide-tolerant animals are also tolerant to ethanol, as measured by a 40% decrease in ethanol-induced sleep times and 15% higher blood alcohol levels upon awakening compared to control mice raised in air. These results suggest a common mechanism for the development of tolerance to both alcohol and nitrous oxide.

Animals↗

Self-reactive T cell hybridomas and tolerance. Same range of antigen dose dependence but higher numbers of self-reactive T cell hybridomas from mice in which self-tolerance has been broken by antiserum treatment.

Mechanisms of self-tolerance of 4-hydroxyphenylpyruvate dioxygenase (HPPD) are explored. It is well established that negative selection based on TCR affinity occurs in the thymus. We have investigated the frequency with which self-reactive T cell hybridomas can be obtained in relation to self-tolerance. Mice immunized with the self-form of HPPD gave rise to T cell hybridomas that were able to recognize self-protein and a synthetic peptide representing the T cell epitope, at higher Ag concentration than was necessary for recognition of allo-protein. The efficiency of negative selection was then reduced by treating neonatal mice with anti-HPPD antiserum. This reduced T cell tolerance of the self-protein, as judged by in vitro proliferation, and enabled self-reactive T cell hybridomas to be generated at a higher frequency. However, the Ag concentration requirements of these hybridomas for the self-protein and the self-peptide remained unaltered. The possibility that these findings reflect an auxiliary mechanism of self-tolerance based on frequencies of self-reactive T cells is discussed.

4-Hydroxyphenylpyruvate Dioxygenase↗

Changes of calcemia and magnesemia during oral glucose tolerance test (OGTT) in relation to glucose tolerance.

We have examined 56 subjects with normal glucose tolerance (NGT), 33 with impaired glucose tolerance (IGT) and 15 diabetics type II (DM) by an oral glucose tolerance test with 75 g of glucose and in addition to glycemia and C-peptide we estimated also calcemia, magnesemia, ionized calcium level (plus corrected ionized calcium level) in all time intervals. The basal glycemia and C-peptide values were different in each examined group, while the magnesium values distinguished only the DM group from the others. The corrected Ca2+ in 60 min decreased significantly in DM, while after 120 min there were no differences in the values. Magnesemia in NGT decreased after 60 and 120 min, while by IGT and DM it rised in both such intervals. The difference between IGT and DM was highly significant. Our findings give evidence about the changes in the distribution of magnesium already at an early stage of decrease glucose tolerance. The importance of this finding for an early IGT diagnostics and its further classification still remains to be definitely evaluated.

Adult↗

[The predictive factor of postpartum impaired glucose tolerance in pregnant women with abnormal glucose tolerance].

To determine factors predictive of impaired glucose tolerance (IGT) in the postpartum period, we examined the maternal and perinatal characteristics of fifty-eight women with abnormal glucose tolerance during pregnancy. A 75g oral glucose tolerance test (OGTT) was performed on the women again around 4 weeks after delivery. Forty women who had a 2-hour plasma glucose level of 140mg/dl or more were defined as IGT in postpartum (group IGT), and the other eighteen women with a 2-hour plasma glucose level lower than 140mg/dl were defined as having normalized glucose tolerance (group N). The women in group IGT weighed more during pregnancy than those in group N (p < 0.05). Insulin secretion during the antepartum OGTT was lower in group IGT than in group N. On the basis of the prepregnant body mass index (BMI) for the obese group (BMI > or = 24), the women in group IGT weighed much more during pregnancy than those in group N (p < 0.03). In the non-obese group, the women in group IGT had higher glucose levels and lower insulin responses than those in group N. We concluded that, in obese women, excessive weight gain during pregnancy is predictive of postpartum IGT, while in non-obese women, the severity of glucose intolerance during pregnancy is related to poor postpartum prognosis.

Adult↗

[The enkephalinase mechanisms of the resistance and tolerance to the analgesic effect of morphine in rats. Differences in the effects of naloxone in morphine-sensitive, morphine-resistant and morphine-tolerant rats].

In morphine-sensitive (s. c. 1.5 mg/kg) Wistar rats i.p. injection of 0.3 mg/kg naloxone either did not change nociception (tail-flick test) or induced hyperalgesia. In morphine-resistant rats 0.2-0.7 mg/kg naloxone injection induced analgetic effect but 1.0 mg/kg induced hyperalgesia. In morphine-sensitive rats chronic morphine administration induced the tolerance and naloxone injection evoked analgetic effect. Morphine inoculation just after naloxone analgesia was over induced analgetic effect. In morphine-resistant and -tolerant rats chronic naloxone administration induced gradual decrease and subsequent disappearance of its analgetic effect and subsequent morphine injections induced analgetic effect for some days. It is suggested that naloxone in morphine-resistant and -tolerant rats with high level of enkephalinase activity functions as its inhibitor. Chronic naloxone administration evoked progressive inhibition of enkephalinase activity that induced the morphine analgetic effect in morphine-resistant and -tolerant rats.

Analgesia↗

'Healthy living' and sulphonylurea therapy have different effects on glucose tolerance and risk factors for vascular disease in subjects with impaired glucose tolerance.

This study was undertaken to determine whether impaired glucose tolerance and associated risk factors for cardiovascular disease can be improved with 'healthy living' by diet and exercise or with sulphonylurea therapy. Patients were recruited by screening subjects with either a family history of type II diabetes, previous gestational diabetes, or a previously raised plasma glucose (5.6-6.6 mmol/l). Impaired glucose tolerance was defined as hyperglycaemia on two separate tests, an achieved glucose level after a glucose infusion test above the 90th percentile of an age-matched normal population (> 9.3 mmol/l) or a fasting plasma glucose above the 95th percentile (> 5.6 mmol/l). Thirty-seven subjects with impaired glucose tolerance were entered into a randomized, prospective study for 6 months with allocations to healthy living or double blind to sulphonylurea (gliclazide 40 mg twice daily) or placebo tablets. The study took place in an out-patient setting, with three times weekly exercise sessions at a Sports Centre. After 6 months the placebo group showed no change in plasma glucose, cholesterol and blood pressure. The subjects receiving gliclazide showed improved glucose levels (mean fasting plasma glucose levels fell from 5.8 to 5.1 mmol/l, p < 0.05) but no significant change in plasma cholesterol or blood pressure. The healthy living group, after exclusion of four non-compliant subjects, showed no change in glucose levels, but a decreased systolic blood pressure (fall in mean from 124 to 116 mmHg, p < 0.05) and plasma cholesterol levels (fall in mean from 5.2 to 4.5 mmol/l, p < 0.01). with an increase in HDL:LDL ratio (rise in mean from 0.39 to 0.46, p < 0.05). Subjects with impaired glucose tolerance may benefit in different ways from gliclazide and healthy living. The metabolic responses to each therapy may help to decrease the risk of developing diabetes and cardiovascular disease.

Adolescent↗

[Carbohydrate tolerance in patients with myocardial infarct according to prednisolone-glucose tolerance test].

Sensitivity to carbohydrates was studied in 100 patients in the subacute period of myocardial infarction by the prednisolne-glucose tolerance test with parallel determination of the content of sugar and pyruvic and lactic acids. Latent, predominantly mild disorders of carbohydrate metabolism were revealed in most patients. Considerably diminished glucose tolerance (diabetic-type sugar curve) was encountered in 12.5% of patients with normal and in 17.9% of those with excessive body weight. The prednisolone-glucose tolerance test and the tests for the content of pyruvic and lactic acids conducted in dynamics showed a statistically reliable increase in the pyruvate level on a fasting stomach and after glucose loads and of the lactate level on a fasting stomach and at the end of the examination. In diabetictype glycemic curves, however, the peak sugar levels were paralleled by normal content of pyruvic and lactic acids. The glycemia indices revealed in dynamics of the prednisolone-glucose tolerance test were significantly higher in patients with an increased blood sugar content in the acute period of myocardial infarction.

Adult↗

Maternal Ig mediates neonatal tolerance in rheumatoid factor transgenic mice but tolerance breaks down in adult mice.

We have recently demonstrated that B cell deletion occurs in the bone marrow of IgHa high affinity anti-IgG2a(a) (RF) transgenic mice. Here we demonstrate via genetic crosses that the source of IgG2a is the mother, thus establishing a transplacental mechanism that ensures tolerance to developmentally expressed Ags. Since maternal IgG can mediate tolerance in young mice, whether tolerance is maintained or, instead, autoimmunity ensues after weaning was investigated. We find that deletion remits abruptly in these RF transgenic mice beginning at 2 to 3 wk postweaning, and some degree of autoreactivity can be observed thereafter for weeks to months. The mechanism of sustained expression of autoreactive RF B cells in normal mice is unclear as yet, but a plausible mechanism is that once self-reactive cells are present, the antibody they secrete markedly reduces the autoantigen levels, presumably allowing further development, rather than deletion, of newly arising B lineage cells. The phenotype of these RF transgenic mice suggests a positive feedback mechanism that tends to perpetuate autoimmunity once it has been established. If such a mechanism were to exist in autoimmune animals, it could have important implications for the establishment and maintenance of B and T cell tolerance in chronic autoimmune diseases.

Aging↗

[Kinetic analysis in the evaluation of glucose tolerance disorders. Evaluation and use of a new method in the diagnosis of glucose tolerance in clinical practice].

A kinetic model of oGTT has been used. This model is characterized by 7 glycaemia collections (0, 30, 45, 60, 120, 180 min). In some cases this is supplemented by determination of C-peptide and insulin values (0, 60, 120 min). This method is very simple and highly useful in clinical practice because it gives information about physiological stimulation by enterohormones, the first glucose passage through liver and next glucose utilization. The first part of the study deals with random error of the kinetic model of oGTT found by repeated calculations (> 10,000 repeatings) of identical initial glycaemias in individual groups (DM, PGT, N). Random error of glucose clearance (the most suitable parameter) ranged within 0.2 -2.1% in individual deviations. Then the identical calculations were made but loaded with glycaemia and with certain error in individual collection intervals (glycaemia 0.1, 0.2, 0.5 mmol/l; collection with 30, 60, 300 s). Random error of the method increased significantly with dispersion variance of glycaemias (maximum 17% with glucose clearance at glycaemia dispersion variance of 0.5 mmol/l); changed time intervals less affected random error quantity. As acceptable and frequent deviation in practice was determined glycaemia dispersion variance of 0.2 mmol/l (corresponding to a total analytical error of glycaemia measurement) and dispersion variance of time intervals of 60 s. At these values, random error of the method increased maximally to 10.3%. Glucose has also a biological variability (not published yet for individual time intervals), the value of random error may be higher but will not achieve half of values of random error obtained at the classical oGTT. The second part of the study deals with comparison of both the tests (classical oGTT, kinetic curve of oGTT) in 126 probands examined at metabolic and diabetologic out-patient department of the Hospital in Sternberk. It can be concluded that using a classical oGTT, 60% of patients were classified into incorrect groups as defined by dynamic results obtained by analysis of the kinetic model. Interesting enough is the fact that almost half of persons who cannot be classified by the classical oGTT had impaired glucose tolerance. Out of them, 15.6% had diabetes mellitus; almost 20% of normal patients also had impaired glucose tolerance. Over 1/5 of persons with impaired tolerance for glucose according to the classical oGTT was found by the kinetic model to have diabetes mellitus. On the contrary, 1/10 of diabetic patients diagnosed by oGTT had normal glucose clearance, over 1/2 of patients had only impaired glucose tolerance. Then in 126 persons random error of the method was again calculated (> 2,000 repeated calculations for each proband) amounting 6.6%.

Glucose Intolerance↗

Phase-I/II study to evaluate dose limiting toxicity, maximum tolerated dose, and tolerability of bendamustine HCl in pre-treated patients with B-chronic lymphocytic leukaemia (Binet stages B and C) requiring therapy.

PURPOSE: Bendamustine hydrochloride, an anti-neoplastic agent with unique mechanism of action, is known to cause impressive remissions in relapsed non-Hodgkin's lymphoma and chronic lymphocytic leukaemia (CLL). Optimal bendamustine dosing for CLL patients had not been finally established and a phase I/II study was conducted to determine the maximum tolerated dose (MTD) and dose limiting toxicity (DLT) of bendamustine. METHODS: The open-label, single-centre phase I/II study was conducted from March 2001 to September 2002 in Sofia, Bulgaria. The 15 study patients were extendedly pre-treated, but fludarabine-naive (3 female, 12 male, 47-72 years of age, 61 years on average). Bendamustine was given at a starting dose of 100 mg/m2 on days 1 and 2 every 3 weeks based on the previous results in lymphoma. RESULTS: Bendamustine was well tolerated in spite of heavy pre-treatment of the study participants. Toxicity corresponded to the known safety profile of bendamustine, with the exception of bilirubin elevation. The level of 110 mg/m2 was established as MTD. A bendamustine dose of 100 mg/m2 is the recommended dose for further clinical investigations. A 4-week interval is recommended to allow for sufficient recovery. Efficacy results confirmed powerful anti-neoplastic activity of bendamustine even in extendedly pre-treated CLL patients. Based on the remission criteria, nine patients were defined as responders (four CRs, two PR, three NC) and two patients as nonresponders to therapy. Four patients were not evaluable for response, because they had received less than three courses bendamustine. After a follow-up period of 15 months, the four patients with CR were still in remission. One patient with PR had relapsed, the other had ongoing response. CONCLUSIONS: Bendamustine is a very active and well-tolerated drug in patients with pre-treated and refractory CLL. Fludarabine-naivity of patients appears to markedly improve their bendamustine tolerability. First-line use of bendamustine is a safe option for CLL-patients requiring treatment, because bendamustine-owing to its unique pharmacodynamics-(1) is highly effective, (2) reasonably safe, and (3) does hardly produce cross-resistance against other anti-neoplastic drugs effective in this indication.

Aged↗

Dopamine-like action of nicotine: lack of tolerance and reverse tolerance.

Rats with unilateral 6-hydroxydopamine lesions of the substantia nigra became briefly sedated and hypothermic after the acute injection of nicotine s.c. (0.4 or 0.8 mg/kg free base). When nicotine was repeated 5 days per week there was rapid tolerance for the sedation and slower tolerance for the hypothermia and the lesioned animals began to rotate ipsiversively after each injection. Stereotypic behavior was also noted. Rats injected with nicotine 5 days per week and nigrally lesioned on the 24th day rotated promptly on their first postoperative injection of nicotine. The nicotinic antagonist, mecamylamine (1.0 mg/kg i.p.), completely blocked the induced rotation. The appearance of rotation did not seem to depend on tolerance to sedation. The direction of rotation indicated enhancement of activity in the intact nigrostriatal system. However, 10 min after the acute injection of 0.8 mg/kg nicotine no change was found in the ratios of dopamine to its metabolites DOPAC and homovanillic acid in the substantia nigra, caudate-putamen, nucleus accumbens, olfactory tubercle, frontal cortex, or ventral tegmental area. Rats given 0.4 or 0.8 mg/kg nicotine 5 days per week and either lesioned prior to nicotine or lesioned during the third week rotated during the sixth week without any sign of tolerance. One day after the 30th injection in intact or lesioned rats the ratios of dopamine to its metabolites did not differ from those in saline controls on either the right or left side of any of the regions examined. There was no evidence of a change in dopamine metabolism after an acute challenge with nicotine or of a sustained change after repeated injection. The possibility remains that repeated nicotine modifies the dopaminergic response to nicotine without causing a sustained change in metabolism.

Animals↗

Analgesic electrical stimulation of the hypothalamic arcuate nucleus: tolerance and its cross-tolerance to 2 Hz or 100 Hz electroacupuncture.

Focal electrical stimulation of the arcuate nucleus of the hypothalamus (ARH) for 5 min (1 session) produced a marked elevation of tail flick latency (TFL) to noxious heat in the pentobarbital-anesthetized rat. Repeated stimulation for a total of 11 sessions at 30 min intervals resulted in a gradual decline in the hypoalgesic action, and this tolerance may last for 7 days. Tolerance to the ARH analgesic stimulation reduced the analgesia produced by low (2 Hz) but not high (100 Hz) frequency electroacupuncture (EA); and tolerance to low frequency EA analgesia attenuated the ARH stimulation-produced analgesia without affecting high frequency EA analgesia. Alternatively, rats tolerant to high-frequency EA analgesia were still sensitive to either the ARH or low-frequency EA stimulation. These results suggest that the ARH stimulation and low-frequency EA administration produced analgesia via a common neural mechanism, supporting our hypothesis put forward previously that the ARH plays an important role in mediating low- but not high-frequency EA analgesia.

Acupuncture Therapy↗

In vitro opioid induced proliferation of peripheral blood immune cells correlates with in vivo cold pressor pain tolerance in humans: a biological marker of pain tolerance.

There is substantial evidence for bidirectional communication between the immune system and the central nervous system, as the cells and signalling molecules of the immune system influence many central nervous system functions, for instance nociception. Opioids, such as morphine, produce analgesia and numerous other central and peripheral effects including sedation and euphoria, while their effects on the immune system are wide-ranging. There is considerable interindividual variability in basal nociception and response to opioids, however, the physiological and biological mechanisms underlying this are unclear. Therefore, we investigated the relationship between the immune system and basal nociceptive thresholds, using the proliferative response of isolated peripheral blood mononuclear cells and cold pressor pain tolerance. Here we show that the percent increase in proliferation of peripheral immune cells from 13 healthy subjects incubated with morphine ex vivo is highly correlated with the subjects' tolerance to noxious cold stimuli (Pearson r = 0.92, P < 0.0001). These pilot data provide evidence of a novel objective biological marker of pain tolerance in humans, which also links the immune and opioid systems with basal pain tolerance.

Adult↗