Depletion of CD8 memory T cells for induction of tolerance of a previously transplanted kidney allograft.

Heterologous immunologic memory has been considered a potent barrier to tolerance induction in primates. Induction of such tolerance for a previously transplanted organ may be more difficult, because specific memory cells can be induced and activated by a transplanted organ. In the current study, we attempted to induce tolerance to a previously transplanted kidney allograft in nonhuman primates. The conditioning regimen consisted of low dose total body irradiation, thymic irradiation, antithymocyte globulin, and anti-CD154 antibody followed by a brief course of a calcineurin inhibitor. This regimen had been shown to induce mixed chimerism and allograft tolerance when kidney transplantation (KTx) and donor bone marrow transplantation (DBMT) were simultaneously performed. However, the same regimen failed to induce mixed chimerism when delayed DBMT was performed after KTx. We found that significant levels of memory T cells remained after conditioning, despite effective depletion of naïve T cells. By adding humanized anti-CD8 monoclonal antibody (cM-T807), CD8 memory T cells were effectively depleted and these recipients successfully achieved mixed chimerism and tolerance. The current studies provide 'proof of principle' that the mixed chimerism approach can induce renal allograft tolerance, even late after organ transplantation if memory T-cell function is adequately controlled.

Long-term islet allograft function in the absence of chronic immunosuppression: a case report of a nonhuman primate previously made tolerant to a renal allograft from the same donor.

UNLABELLED: Development of mixed chimerism by donor bone marrow transplantation (DBMT) has led to long-term tolerance of solid organ allografts in nonhuman primates. As an initial attempt to extend this approach to cellular transplant, islet transplant from the same donor was attempted in the recipient previously made tolerant to a kidney allograft. METHODS: After the conditioning with ATG, total body irradiation, thymic irradiation, and splenectomy, DBMT was performed followed by 4 weeks of cyclosporine. Kidney transplantation and native nephrectomies were subsequently performed on day 89. After 2.8 years of DBMT, diabetes was induced by streptozocin (STZ) and islets from bone marrow and kidney donor were transplanted without immunosuppression. RESULTS: After DBMT, the recipient developed chimerism and no evidence of kidney rejection for more than 1000 days. STZ induced diabetes was reversed after the islet transplantation. Islet biopsies demonstrated insulin staining without rejection. Although the recipient became diabetic 300 days after islet transplantation, viable transplanted islets were found in the liver and under the kidney capsule without any evidence of rejection. CONCLUSION: Tolerance with a nonmyeloablative conditioning can allow successful pancreatic islet transplantation without immunosuppression. Because no histological evidence of rejection was identified, recurrent diabetes is presumed to be inadequate islet mass.

Orexin-A and leptin change inversely in fasting non-obese subjects.

OBJECTIVE: Leptin, neuropeptide-Y (NPY) and orexin are peptides regulating energy metabolism and appetite control. NPY and orexin are mainly found in the central nervous system and they have also recently been found in the peripheral nervous system. We investigated how fasting affects changes in circulating concentrations of these peptides and their association with nutritional and metabolic parameters in humans. DESIGN AND METHODS: Ten non-obese female patients with psychosomatic disorders fasted for 7 or 10 days. Blood samples were collected at 0800 h before fasting, on the 3rd and 7th days during the fast (with an additional sample taken on the 10th day when the fasting continued for 10 days) and on the 3rd and 7th days of refeeding. We measured blood concentrations of orexin-A, NPY, leptin, adrenocorticotropin, cortisol, insulin, C-peptide, glucose, and beta-hydroxybutyrate. RESULTS: Body mass index and plasma leptin concentrations concomitantly and significantly decreased during fasting, whereas serum orexin-A concentrations significantly increased and were negatively correlated with plasma leptin concentrations. Plasma NPY concentrations decreased slightly but were not significantly different from the prefasting values, and no significant relationship with leptin or orexin-A was found. Orexin-A and leptin concentrations showed a significant inverse correlation with serum glucose, insulin, C-peptide, and beta-hydroxybutyrate concentrations. Only changes in plasma leptin concentrations showed a significant negative correlation with serum cortisol concentrations. All the measured indices which changed during fasting returned to the prefasting concentrations by the 7th day of refeeding. CONCLUSION: Peripheral orexin-A and leptin concentrations inversely change during fasting, which is significantly correlated with energy metabolism in humans.

Antibiotic use during infancy promotes a shift in the T(H)1/T(H)2 balance toward T(H)2-dominant immunity in mice.

BACKGROUND: Recent epidemiologic studies indicate that antibiotic use in infancy may be associated with an increased risk of development of atopy; however, its precise mechanism remains to be elucidated. OBJECTIVE: The purpose of this study is to clarify whether one such antibiotic, kanamycin, affects the T(H)1/T(H)2 balance. METHODS: BALB/c mice at 3 and 52 weeks of age were orally administered 600 mg/d kanamycin sulfate for 7 consecutive days. Blood samples were collected on weeks 0, 10, 18, and 26 after the cessation of kanamycin treatment, and the effect of the kanamycin treatment on the T(H)1/T(H)2 balance was evaluated on the basis of both the in vivo antibody levels and the in vitro splenocyte cytokine secretion pattern. RESULTS: The administration of kanamycin increased the serum levels of total IgG1 and IgE while decreasing the serum IgG2a levels. Moreover, when spleen cells were stimulated with immobilized anti-CD3 antibody in vitro, such kanamycin treatment enhanced the in vitro IL-4 secretion while reducing the in vitro IFN-gamma secretion. The basal IL-12 p70 secretion levels of splenic dendritic cells in the kanamycin-treated mice were lower than those in the control mice, although no significant difference was seen in IL-12 p40 levels between either group of mice. CONCLUSION: These results suggested that antibiotic use during infancy may indeed quantitatively disturb, qualitatively disturb, or both the intestinal microflora and thereby prevent postnatal T(H)1 cell maturation, thus resulting in a T(H)2-polarized immune deviation.

Influence of short-term repeated fasting on the longevity of female (NZB x NZW)F1 mice.

Caloric restriction in rodents is well known to retard the rate of aging, increase mean and maximum life-spans, and inhibit the occurrence of many age-associated diseases. However, little is known about the influence of short-term repeated fasting on longevity. In this study, female (NZB x NZW)F1 mice were used to test the physiological effect of short-term repeated fasting (4 consecutive days, every 2 weeks). The results showed that fasting mice survived significantly longer than the full-fed mice, in spite of the fasting group having a heavier body weight than the control group. Mean survival times for fasting and control mice were 64.0+/-15.3 and 47.9+/-9.4 weeks, respectively. Short-term repeated fasting manipulation was also effective on the prolongation of life-span in autoimmune-prone mice.

The restraint stress-induced reduction in lymphocyte cell number in lymphoid organs correlates with the suppression of in vivo antibody production.

In this study, we examined the effects of restraint stress on some immune parameters such as the in vivo antibody levels, cytokine production, and lymphocyte cell number in the spleen or mesenteric lymph node (MLN). BALB/c mice were thus injected intraperitoneally 2-times with OVA absorbed into alum on days 0 and 21. Before the first injection, the animals were either restrained for 12 h (stress group) or returned to their home cage (control group). Exposure to stress resulted in a reduction in the serum levels of anti-OVA IgE, IgG1, and IgG2a. In addition, stress also caused a decrease in the IL-4 and IFN-gamma levels in the spleen or mesenteric lymph node cell culture supernatants. Furthermore, exposure to stress resulted in a decrease in the splenic and mesenteric lymphocyte cell number when examined immediately after the cessation of stress. This decrease persisted for at least 12 h after the termination of stress and thereafter disappeared 24 h after stress. The stress-induced reductions in antibody and cytokine production occurred only when antigen was given either immediately or 6 h after stress, but not when antigen was given 24 h post stress. These results thus suggest that the restraint stress-induced change in lymphocyte cell number in the spleen or MLN closely correlates with the altered antibody and cytokine levels.

Alterations in lymphocyte subsets and pituitary-adrenal gland-related hormones during fasting.

We investigated changes in the immunoendocrine system during fasting. Ten hospitalized patients aged 14-46 y with psychosomatic disorders fasted for 7 or 10 d. Blood samples were collected before and on days 3 and 7 of the 7-d fasts. When fasting continued to 10 d, an additional sample was taken on day 10. We measured blood cellularity (white blood cells and total lymphocytes), the total number and percentage of lymphocyte subsets (CD2, CD3, CD4, CD8, and CD19), natural killer (NK) cell activity, cytokines (interleukin 1 beta, interleukin 2, interleukin 6, granulocyte-macrophage colony stimulating factor, tumor necrosis factor alpha, and interferon gamma), and soluble interleukin 2 receptors. Corticotropin, cortisol, and dehydroepiandrosterone sulfate (DHEAS) concentrations were also determined. Although the total number of lymphocytes decreased during fasting, NK cell activity increased significantly. Plasma cortisol and DHEAS concentrations also increased significantly whereas changes in corticotropin concentrations were not significant. The total number and percentage of CD4 cells decreased significantly during fasting but no other lymphocyte subsets changed significantly. The percentage of CD4 cells was negatively correlated with cortisol concentrations during fasting. No detectable changes occurred in cytokines or soluble interleukin 2 receptors during the study. All measured immunoendocrine values that changed during fasting returned to prefasting values during the refeeding period. These findings indicate that fasting affects immune variables such as T cell subsets and NK cell activity at least in part through changes in adrenal gland-related hormones.

Restraint stress causes tissue-specific changes in the immune cell distribution.

In this study, the effect of restraint stress on alterations in the immune cell distribution was examined in bone marrow, liver, thymus, and spleen. In bone marrow, stress induced a striking increase in both the proportion and number of CD3+CD4+, CD3+CD8+, B220brightIgM+, CD3-IL-2R beta + and CD3intIL-2R beta + cells. Such an increase was partially reversed by pretreatment with RU-486, a steroid receptor antagonist, while it was profoundly enhanced by either sympathectomy with 6-hydroxydopamine hydrobromide or by a beta-adrenergic blockade with propranolol, a beta-adrenergic receptor antagonist; this suggests that corticosteroids and catecholamines may act in opposition with regard to such an immune-cell accumulation in bone marrow. In the liver, stress decreased the proportions of CD3intIL-2R beta +, CD3-IL-2R beta +, and B220brightIgM+ cells, while it increased the proportion of CD3brightIL-2R beta-cells, thus demonstrating that different subpopulations were differentially affected. In the thymus and spleen, stress only slightly affected the proportions of lymphocyte subpopulations, although both tissues showed a drastic reduction in the number of lymphocytes. Taken together, these results suggest that restraint stress induces tissue-specific changes in the immune-cell distribution.

[A case of vocal cord dysfunction diagnosed as bronchial asthma, that was improved by psychosomatic therapy].

A 19-year-old psychologically disturbed female was admitted to our hospital because of intractable dyspnea and wheezing for twenty two months. She had been diagnosed as having asthma and had received several medications including steroids before admission to our hospital. According to our clinical observations, she was considered not to have asthma because neither FEV1.0 nor PaO2 was decreased during the periods of wheezing and FEV1.0 was not changed by inhalation of bronchodilators, although she had a family history of allergy and false bronchial hyperreactivity by asthograph. On physical examination, diffuse wheezing was heard mainly over the larynx during inspiration at the times of dyspnea. There was flattening of the inspiratory flow-volume loop during wheezing. Bronchoscopy performed during an attack confirmed that wheezing was due to adduction of the vocal cords throughout the respiratory cycle. Therefore, this case was diagnosed as vocal cord dysfunction. Her symptom was considered to be a form of conversion reaction derived from her unhappy past history. Following psychosomatic therapy, all of her medications became unnecessary. She understood the mind-body relationship of her condition, and learned to achieve self-control.

[Graduated psychosomatic treatment and quality of life in asthmatics].

We investigated the quality of life (QOL) in 72 patients with bronchial asthma who are under our gradational psychosomatic treatment (GPT) by means of an 11-item questionnaire. The results were summarized as follows: 1) Ninety-two percent of the subjects showed a good understanding of mind-body relations and modified their stressful adaptive patterns. 2) Asthmatic symptoms improved in 86 percent of the subjects. Other symptoms also improved in 72 percent of the subjects. 3) In 81 percent of the subjects, their daily life improved. Furthermore 96 percent of the subjects obtained some advantages through GPT. 4) There were various improvements in psychological states, personal relations and life style in most of the subjects. 5) The attitudes of their families toward the patients improved in 46 percent of the subjects, however 71 percent of the families developed a better understanding of the cause of asthma from the psychosomatic point of view. 6) Most of the subjects with moderate or severe symptoms were considered to have reduced the grade of severity of their symptoms from the doctor's standpoint.

Effects of the biopsychosocial approach (BPSA) on exercise-induced asthma (EIA).

The Biopsychosocial Approach (BPSA) is a treatment program for allergic patients which includes therapy for psychological, behavioral and social factors as well as for physical problems, following basic principles of psychoneuroimmunology. BPSA was applied to patients with bronchial asthma and favorable results were obtained. The mechanism of the therapeutic effects of BPSA included normalization of the patient's autonomic nervous function, levels of blood histamine, and circadian rhythm of lymphocyte activity. BPSA was also used in patients with exercise induced asthma (EIA) and the same parameters were evaluated. Results showed that patients with EIA recovered physiological homeostasis after BPSA therapy normalized blood levels of histamine and substance P (SP), skin reactions to histamine and SP, and autonomic nervous function. We conclude that BPSA is effective for treating patients with EIA.

Long-term follow-up investigation of the effects of the biopsychosocial approach (BPSA) to bronchial asthma.

The recent rapid increase in the number of allergic patients is becoming a social problem. Studies of the causes of this phenomenon involve various fields, with much attention focused on finding new antigens in food, air, articles encountered in daily living, etc. Recent studies of psychoneuroimmunology (PNI) also suggest a strong influence of emotions on allergic reactions. The number of allergic patients is increasing in all civilized countries without exception, and the stress prevalent in modern civilized society is related to this increase. Modern allergology does not yet have sufficient countermeasures for such stress states. We applied a biopsychosocial approach (BPSA) to treatment programs for allergic disease, incorporating treatment of physical and psychosocial problems en bloc. We studied the long-term effects of BPSA therapy on 82 patients who were treated for more than 3 months in the hospital and were examined 2 to 3 years after discharge. Results showed that more than 80% of patients maintained improvement and 45% of those with intractable asthma were able to withdraw from steroid hormones. BPSA achieved better results than those with standard medication administered only to the body. Improvements after treatment included physical changes, normalization of MV (microvibration) type, decreased levels of plasma histamine, and normal circadian rhythms of lymphocyte subsets. These changes reflect part of the physical mechanisms by which BPSA improves asthma symptoms. From a psychological view point, the patients' feelings, personal relations, behavior, etc. were changed after BPSA, allowing a new life style and improved QOL. It is important for asthma patients to maintain good overall condition over long periods. After BPSA, 80% of our patients were able to do so. It is difficult for the therapist to approach asthma from different aspects at once, including biological, psychological, and social, so we developed a five-stage program of BPSA therapy and found that this obtained favorable results.

[Psychosomatic study of inhalation therapy in patients with bronchial asthma. II. Relationship between subjective self assessment of dyspnea and peak expiratory flow rates].

The degree of dyspnea when using metered-dose inhalers (MDIs), and the relationship between subjective self assessments and objective peak expiratory flow rates were examined by means of visual analogue scales and mini wright peak flow meters in 16 subjects with asthma. Three subjects who were neurotic in character were found to have a poor perception of the degree of airway obstruction after inhalation compared with objective changes in their condition. Two subjects, one neurotic and one alexithymic, assessed the degree of dyspnea to be worse after inhalation, when objectively there were few changes. However, one neurotic subject perceived an improvement in his condition when, in fact, there was little change. Furthermore, some severe cases used MDIs in the rogressive state of airway obstruction. In conclusion, we must pay attention to patients' characters when determining treatment in order to prevent overuse of MDIs or delays in treatments including the use of MDIs.

[Psychosomatic study of inhalation therapy in patients with bronchial asthma. I. Evaluation by questionnaire].

We investigated the use of metered-dose beta agonist inhalers (MDIs) by means of 17-item questionnaires from the standpoint of psychosomatic medicine in 56 subjects with bronchial asthma. The results can be summarized as follows: 1) Many asthmatics with neurotic characters used MDIs frequently; Thirteen percent of them suffered adverse effects. The number of times the inhalers were used increased in proportion to the severity and the duration of illness. Fifteen percent of the subjects used MDIs frequently because of anxiety. 2) Fifty percent of the subjects, most of them had neurotic or alexithymic characters, used MDIs without suffering from dyspnea or wheezes; the subjects who used MDIs because of anxiety increased in proportion to the duration of illness. 3) Seventy eight percent of the subjects were anxious about asthmatic attack when they did not carry any MDI; sixty two percent of the subjects, most of them had neurotic or alexithymic characters, actually experienced asthmatic attack. 4) Seventy three percent of the subjects used MDIs secretly; sixty percent of the subjects did not like to inhale in the presence of others. 5) There were some subjects who acted incorrectly on or after asthmatic attack when MDIs were not effective.

Changes in T-cell subpopulations induced by autonomic neurotransmitters and hormone.

In our previous studies, we found that changes in T-cell subpopulations were induced by restraint stress. To determine how this influence on the immune system was mediated pharmacologically, we studied the effects of autonomic agonists and antagonists and hydrocortisone on T-cell subpopulations in C3/H mice. These agents induced changes in the subpopulations of T-cells and modulated the receptors on T-cells. These drugs, however, did not have the same effect as restraint stress. This suggests that the effect of restraint stress on the immune system is mediated by other mechanisms. We found autonomic agents and hydrocortisone to affect differently T-cells in various organs. In general, acetylcholine and hydrocortisone affected T-cells in the thymus and adrenaline affected those in the spleen. By the double staining method, the maturity of these T-cells was found to be affected by these drugs. Remarkable changes were as follows. Thy 1, 2-positive T-cells were decreased in the blood and increased in the spleen while Lyt 2-positive T-cells were increased in the spleen by adrenaline. Thy 1, 2-positive T-cells were decreased in the thymus and increased in the blood while L3T4-positive T-cells and Lyt 2-positive T-cells in the blood were increased by acetylcholine. L3T4-positive T-cells were decreased in the thymus by atropine. Finally, Lyt 2-positive T-cells in the thymus were decreased and L3T4-positive T-cells in the blood were increased by hydrocortisone.(ABSTRACT TRUNCATED AT 250 WORDS)