Suppressive effect of acupuncture on delayed type hypersensitivity to trinitrochlorobenzene and involvement of opiate receptors.

We reported previously that electroacupuncture (Acu) at an acu-point equivalent to GV-4 in mice either enhanced or suppressed the delayed type hypersensitivity (DTH) to 2,4,6-trinitrochlorobenzene (TNCB, picryl chloride) depending on the time of treatment. We report here the suppression of the efferent phase of DTH to TNCB by Acu in mice. In 7- to 9-week-old male BALB/c, C57BL/6 and ddY mice, significant suppression of the DTH was observed when Acu had been applied once per day for three consecutive days before TNCB challenge. When Acu had been applied a single, significant suppression also occurred. Application to another point (at a middle area of the femoral muscle) failed to suppress the DTH to TNCB. This Acu-evoked DTH suppression was blocked dose-relatedly by pretreatment of systemic naloxone hydrochloride, indicating that opioid receptor-mediated mechanisms are involved in this immune response.     

The effects of the delayed type immune response upon the host

A consistent accumulation of experimental evidence, most of it published long ago, clearly shows that a constant consequence of the immune response elicited by the Koch bacillus is that the individual becomes susceptible to be injured or killed by the tuberculous antigen, which is totally innocuous to the virgin subject. This fact led Rich to conclude that throughout the natural history of tuberculosis, as soon as the immune reaction is activated, the tuberculo-protein poured out by the germ mass will exert its detrimental capabilities manifested as the constitutional symptoms of the disease. The pathogenicity of the specific antigen in the tuberculous-type immune reaction may be extrapolated to other states that elicit the same response, including cancer and tissue transplantation. Unexplained phenomena such as the general symptoms of many neoplasias, as well as the systemic manifestations of graft rejection, may then be attributed to the shedding of antigen from the foreign cell mass into the sensitized host.     

Secondary delayed type hypersensitivity to H-2 subregion-coded alloantigens

Secondary type delayed type hypersensitivity (DTH) in mice against class I alloantigens or non-H-2 alloantigens is characterized by an earlier appearance of DTH reactivity after booster immunization compared with the development of DTH reactivity after primary immunization. In contrast to the primary and secondary DTH against class I or non-H-2 alloantigens, the development of secondary DTH against class II alloantigens or a set of alloantigens that includes class II alloantigens is not faster than the development of primary DTH. Thus, primary and secondary immunization with class II alloantigens prevents secondary type DTH reactivity to simultaneously administered class I alloantigens and non-H-2 alloantigens.     

Asymmetry of delayed type hypersensitivity reaction in mice

The intensity of delayed type hypersensitivity reactions in the left and right paws was studied in mice divided into left- and right-pawed by the motor asymmetry of the brain. The reaction was more pronounced in the left paw in all animals irrespective of motor asymmetry. Motor asymmetry of the brain hemispheres little contributed to the manifestation of differences in the delayed type hypersensitivity reactions in the left and right paws. The authors concluded that asymmetry of cellular immunity is determined by functional asymmetry of cells in regional lymph nodes.     

The beneficial effects of adjunctive recombinant human interleukin-2 for multidrug resistant tuberculosis

Introduction

Multidrug-resistant tuberculosis (MDR-TB) is a hard-to-treat disease with a poor outcome of chemotherapy. In the present study, the efficacy and safety of recombinant human interleukin-2 (rhIL-2) were investigated in patients with MDR-TB.

Material and methods

Fifty culture-confirmed patients with MDR-TB were included. Twenty-five patients were randomly assigned to the trial group (injection of 500 000 IU of rhIL-2 once every other day at the first, third, fifth and seventh months in addition to standard multidrug therapy) and another 25 patients to the control group with standard multidrug therapy. All patients were monitored clinically, and T-cell subsets were analyzed by flow cytometry.

Results

The rates of sputum negative conversion and X-ray resolution in the trial group were higher than those of the control, and the improvements were significant by completion of treatment. In addition, CD4⁺CD25⁺ T cells in the controls rose gradually during treatment. The levels at the end of the seventh month were significantly higher than before, which were also significantly different when compared with those from the trial group at the same time. However, there were no such changes associated with treatment in the trial group. No significant differences appeared in other T cell subsets.

Conclusions

Exogenous IL-2 in the present regimen improves immunity status. Adjunctive immunotherapy with a long period of rhIL-2 is a promising treatment modality for MDR-TB.     

Atopic allergy and delayed type hypersensitivity in Estonian children

BACKGROUND: A shift in the balance ofT helper (Th) cell subsets towards a polarized Th2 population is generally accepted to occur in atopic disease, however, both Th1 and Th2 disorders have increased over the past decades in Western communities. OBJECTIVE: The aim of our study was to investigate delayed type hypersensitivity (DTH) response in atopic and non-atopic children in a population with a low prevalence of allergic disorders. METHODS: Skin prick tests (SPT) were performed with fresh egg white and extracts of five inhalant allergens, i.e. cat, dog, house dust mite (Dermatophagoides pteronyssinus), birch and timothy, and DTH response was evaluated by Multitest CMI in 72 Estonian 4- to 6-year-old children. RESULTS: The frequency of response to diphtheria was significantly increased in SPT-positive children (55% vs. 26%, chi2 = 5.5; P = 0.038). The induration to diphtheria (2.4 +/- 0.5 vs. 0.9 +/- 0.2 mm; P = 0.004), and tetanus (3.5 +/- 0.6 vs. 2.1 +/- 0.3 mm; P = 0.025) was significantly greater in the SPT-positive children. The cumulative size of induration in the positive DTH tests was significantly greater in the SPT-positive children (9.0 +/- 1.2 vs. 5.2 +/- 0.6 mm, P = 0.01). CONCLUSION: In this group of children our findings do not support the hypothesis of an immune deviation with decreased Th1 and increased Th2 responses leading to atopic disease, but rather a process of immune modulation whereby both Th1 and Th2 responses are increased in atopic subjects.     

Immediate and delayed type hypersensitivity to malathion.

Between December 1989 and June 1990, 1,874 reports of alleged malathion application related illness from repeated spraying of a mixture of malathion corn syrup bait to eradicate a Mediterranean fruit fly infestation in Southern California were received by the Toxics Epidemiology Program of Los Angeles County. Among these complaints were 47 reports of urticaria, 38 reports of angioedema and 213 reports of a nonspecific skin rash. In order to determine whether these alleged skin reactions were the result of an immediate or delayed type of hypersensitivity reaction to malathion or to the corn syrup bait we studied ten subjects referred for testing by the local health department. All ten subjects had no reaction on patch testing. One child exhibited a positive reaction to the bait and one child had irritant reactions to malathion and to the bait. This study documented one case of a possible immediate IgE reaction to malathion bait. Due to the low participation rates in this study, no specific conclusions concerning the rate of sensitivity in the population can be drawn, although it appears that such reactions are uncommon.     

Effects of administration of recombinant human interleukin-2 in dogs

The clinical and immunohaeamatological effects of recombinant humen interleukin-2 (rhIL-2) administration were evaluated in normal dogs. Three groups of three dogs per group were administered rhIL-2 subcutaneously at a dose of 6 × 10⁴ IU, 6 × 10⁵ IU, or 6 × 10⁶ IU/kg once daily for five consecutive days. Toxic clinical signs were limited primarily to diarrhoea, the severity of which, was dose dependent, with resolution within 7 days of the last rhIL-2 injection. Marked circulating eosinophilia occurred in dogs of the two highest dose groups and transient rise in blood lymphocyte numbers occurred in dogs given the highest dose of rhIL-2. The most significant immunological effects were elevated in vitro conA and pokeweed mitogen-stimulated lymphocyte blastogenic responsiveness in the highest dose group and dose-dependent elevation of antigen-specific antibody (IgG and IgM) production. Peak relative antibody production was markedly elevated, as compared to controls, in dogs administered 6 × 10⁵ IU, 6 × 10⁶ IU rhIL-2/kg.     

国产虫草素(cordycepin)抗小鼠迟发型超敏反应的实验研究

目的：研究国产虫草素抗小鼠迟发型超敏反应的作用及其免疫机理。方法：用2，4-二硝基氯苯（2，4-DNCB）对昆明种小鼠皮肤致敏和诱发，制成迟发型超敏反应模型。以国产虫草素溶液作为实验组，分为两个剂量组（实验组1：12mg／kg；实验组2：16mg／kg），以生理盐水作为阴性对照组，所有溶液均为间隔24小时（最后一次间隔为4小时）重复腹腔注射给药。计算激发后各组小鼠左右耳耳廓肿胀厚度差、耳廓增重值及肿胀度，进行统计分析。并经过HE染色观察虫草素对小鼠致炎耳炎症病理变化及脾脏病理变化的影响。结果：两种给药剂量的国产虫草素溶液对模型鼠红斑，水肿。渗出的接触性皮炎损害有明显抑制作用，可减轻由于充血、水肿等炎症反应导致的局部皮损增厚及重量的增加。与生理盐水对照组相比。两实验组虫草素溶液的抗炎作用均有显著性差异（厚度差：P〈0．0001；肿胀度：P〈0．05）；两实验组间的抗炎作用也有显著性差异（厚度差及肿胀度均为P〈0．05），且与给药剂量相关。三组小鼠脾指数未见明显差异（P〉0．05），脾脏组织病理变化未见明显差异。结论：虫草素以24小时间隔（最后一次给药间隔为4小时）经腹腔注射后，可能通过其免疫调节作用对迟发型超敏反应引起的小鼠接触性皮炎发挥明显的抑制效应，该效应与给药剂量相关，同时对脾脏组织未见明显毒性作用。     

The induction of delayed type hypersensitivity to dinitrochlorobenzene in the chicken

Delayed hypersensitivity to DNCB in chickens is a dose-dependent reaction. It could be induced by a single painting of the skin with 1 mg of DNCB/bird without adjuvant and could be elicited at a challenge site with 50 mug DNCB. The sensitisation lasted for several weeks, and was mediated by sensitised lymphoid cells which required the presence of autologous cells that adhered to plastic for the reaction to be transferred to other allogeneic birds. Doses above 1 mg/bird induced some form of suppression. The migration inhibition test could be used as an in vitro method to measure the DTH response and correlated with the DTH skin response. The histological appearance of skin lesions was characterised by the infiltration of a large number of granulocytes especially heterophils and some eosinophils, as well as large numbers of monocytes.     

The effects of cyclosporin A on leucocyte infiltration and procoagulant activity in the mouse delayed hypersensitivity response in vivo

A model of delayed type hypersensitivity (DTH) to methylated bovine serum albumin was established and characterised in the mouse pleural cavity. Fluid exudate formation was delayed, peaking around 24 h and diminishing by 48-72 h. Exudate leucocyte accumulation was also delayed, rising by 24 h and reaching maximal levels between 24-72 h. Significant PMN leucocyte infiltration was observed early on, although macrophages dominated the reaction from 24 h onwards. Small numbers of lymphoid cells were also observed. Treatment with cyclosporin A (CsA) at 100 mg/kg p.o. around the time of antigen challenge significantly reduced fluid exudate volume but consistently failed to reduce either leucocyte infiltration or the PMN: mononuclear cell ratio throughout the reaction. CsA treatment enhanced DTH exudate leucocyte procoagulant activity (PCA) although leucocyte morphology remained unaltered. The development of carrageenan edema in the hind paws of mice was unaffected by CsA treatment (50-200 mg/kg p.o.). These results suggest that (1) CsA selectively suppresses DTH vascular permeability responses without affecting leucocyte accumulation and (2) CsA does not affect the lymphokine-mediated induction of leucocyte PCA associated with DTH reactions in vivo.     

骨髓Sca-1+间充质干细胞移植减轻小鼠迟发型超敏反应

 目的 探讨骨髓Sca-1+间充质干细胞（BM-MSCs）在迟发型超敏反应（DTH）小鼠中的免疫调控作用。方法 小鼠随机分为对照组及MSCs注射组。注射组于0、2及6 d腹腔注射Balb/c小鼠Sca-1+ BM-MSCs,对照组注射生理盐水。所有鼠在7和14 d时分别于背部皮下与足跖注射C57BL/6小鼠脾细胞。24 h后用千分尺测量小鼠足跖的肿胀,ELISA法测外周血细胞因子及FACS法测脾脏免疫细胞比例。结果 注射组小鼠足跖的肿胀程度（0.368 ? 0.126 mm）显著轻于对照组（0.731 ? 0.111 mm,p < 0.01）。注射组外周血中IL-10和TGF-β含量显著高于对照组（p < 0.05）;而IL-12和TNF-α水平显著低于对照组（p < 0.05）。注射组小鼠脾脏调节型T细胞、树突细胞的比例明显高于对照组。结论 BM-MSC通过上调抑炎因子和调节型免疫细胞的数量而减轻DTH小鼠的免疫反应。     

Sca-1 + bone marrow mesenchymal stem cells transplantation for the alleviation of delayed type hypersensitivity in mouse

目的 探讨骨髓Sca-1+间充质干细胞(BMSCs)在迟发型超敏反应(DTH)小鼠中的免疫调控作用.方法 BALB/c小鼠随机分为对照组及MSCs注射组.注射组于0、2及6d腹腔注射Sca-1+ BMSCs,对照组注射0.9％氯化钠注射液.所有小鼠在7和14 d时分别于背部皮下与足跖注射C57BL/6小鼠脾细胞.24h后用千分尺测量小鼠足跖的肿胀,ELISA法测外周血细胞因子及FACS法测脾脏免疫细胞比例.结果 注射组小鼠足跖的肿胀程度[ (0.368 *0.126) mm]显著轻于对照组[(0.731±0.111) mm,P＜0.01].注射组外周血中IL-10和TGF-β含量显著高于对照组(P＜0.05);而IL-12和TNF-α水平显著低于对照组(P＜0.05).注射组小鼠脾脏调节型T细胞和树突细胞的比例均明显高于对照组(P＜0.05).结论 BMSC通过上调抑炎因子和调节型免疫细胞的数量而减轻DTH小鼠的免疫反应.     

A new method of eliciting delayed hypersensitivity in the mouse abdominal cavity

A new method of determining delayed hypersensitivity quantitatively was investigated in mice. Mice were sensitized with 150 micrograms of ferritin and, 3 weeks later, antigen challenge was performed by implanting a sponge containing antigen in the abdominal cavity. Cells accumulated in the sponge markedly increased in number for 24-72 h after the challenge; mononuclear cells predominated by 48 h. When sensitized lymphocytes were transferred passively to a normal recipient, marked cell accumulation in the sponge was found 48 h after the challenge. Immunological specificity was confirmed in animals sensitized to antigen and receiving passive transfer of sensitized cells. Strain differences in this reaction were observed. Cortisone (20 mg/kg for 6 days before challenge) significantly decreased cell accumulation. Delayed hypersensitivity was also elicited in the ear of sensitized animals. Extracts of sponges removed from antigen-challenged mice had macrophage chemotactic activity.     

Psychological modulation of the delayed type hypersensitivity skin test.

Considerable data have demonstrated that psychological states can influence the immune system in animals. Whether human immune function can be intentionally modulated by the central nervous system is unknown. This article presents data from two studies that sought to demonstrate intentional modulation of the immune system by psychological interventions. It also discusses the methodological complexities involved with this type of research in humans.     

Differential effects of interleukin-12, interleukin-15, and interleukin-2 on human immunodeficiency virus type 1 replication in vitro.

Cytokines may have clinical utility as therapeutic agents for human immunodeficiency virus type 1 (HIV-1) infection and as an adjuvant for vaccines. The effect of interleukin-12 (IL-12) and IL-15 on in vitro HIV-1 replication was investigated. IL-12 and IL-15 at doses up to 10 ng/ml had little effect on basal HIV-1 p24 antigen production by chronically HIV-infected T (ACH-2) and monocytic (U1) cell lines. For ACH-2 cells stimulated with phorbol 12-myristate 13-acetate (PMA; 50 ng/ml), IL-12 and IL-15 significantly increased p24 antigen production by 20 and 30%, respectively (n = 6). In contrast, IL-12 and IL-15 (10 ng/ml) treatment of PMA-stimulated U1 cells decreased p24 antigen production by 16 and 15%, respectively (n = 6). We next studied the effect of IL-12 and IL-15 on HIV-infected peripheral blood mononuclear cells (PBMCs). In 10 HIV-seropositive patients' PBMCs cocultured with mitogen-activated HIV-seronegative donor cells, two patterns of p24 antigen production were observed in response to IL-2: low (p24 antigen production < 10(3) pg/ml; n = 8) and high (p24 antigen production > 10(3) pg/ml; n = 2) response. For the low-response pattern, IL-12 and IL-15 increased viral replication by 97-fold and 100-fold, respectively (P = 0.05 and 0.004, respectively). For the high-response pattern, both IL-12 and IL-15 suppressed HIV replication. The effect of IL-2, IL-12, and IL-15 on acute in vitro infection by HIV-1JRCSF was also examined. IL-12 did not increase p24 antigen production above basal levels while IL-2 and IL-15 significantly enhanced p24 antigen production (by approximately 2-fold). In conclusion, IL-12 and IL-15 may have differential effects on latent and acute HIV infection, and their ability to enhance HIV production may depend on cell activation. Thus, the use of these cytokines may be dictated by the clinical state of the patient.     

Dose-dependent effects of recombinant human interleukin-6 on the pituitary-testicular axis.

Inflammatory cytokines are soluble mediators of immune function that also regulate intermediate metabolism and several endocrine axes. To examine the effects of interleukin-6 (IL-6), the main circulating cytokine, on the hypothalamic-pituitary-testicular axis in men, we performed dose-response studies of recombinant human IL-6 (rHuIL-6) in normal volunteers. Increasing single doses of IL-6 (0.1, 0.3, 1.0, 3.0, and 10.0 microg/kg body weight) were injected subcutaneously into 15 healthy male volunteers (3 at each dose) in the morning. We measured the circulating levels of testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), and sex hormone binding globulin (SHBG) at baseline and then at 24 h, 48 h, and 7 days after the IL-6 injection. LH and FSH levels were also measured half-hourly for the first 4 h after the IL-6 injection. All IL-6 doses were tolerated well and produced no significant adverse effects. Mean peak plasma IL-6 levels achieved after IL-6 administration were 8 +/- 1, 22 +/- 5, 65 +/- 22, 290 +/- 38, and 4050 +/- 149 pg/ml, respectively for the five doses. We observed no significant changes in plasma testosterone levels after the two smaller IL-6 doses. The three higher IL-6 doses, however, caused significant decreases in testosterone levels by 24 h, which persisted at 48 h and returned to baseline by 7 days. The higher testosterone suppression was after the 3.0 microg/kg dose, making the dose-response curve bell-shaped. There also appeared to be small but not significant increases in LH levels after the three higher IL-6 doses, which were not acute and seemed to follow temporally the testosterone decreases. The concurrent plasma levels of FSH and SHBG were not appreciably affected by any IL-6 dose. In conclusion, subcutaneous IL-6 administration, which caused acute elevations in circulating IL-6 levels of a similar magnitude to those observed in severe inflammatory and noninflammatory stress, induced prolonged suppression in testosterone levels in healthy men without apparent changes in gonadotropin levels. This suggests that IL-6 might induce persistent testicular resistance to LH action or suppression of Leydig cell steroidogenesis or both, with potential adverse effects on male reproductive function.     

Cells that mediate NK like cytotoxicity are present in the human delayed type hypersensitivity response.

By inducing delayed type hypersensitivity (DTH) responses under previously formed skin blisters we determined that cells which mediate natural killer (NK) like cytotoxicity are present in the DTH response in man. Similar levels of killing were not present in cells obtained from skin blisters not associated with positive DTH responses. The DTH response associated killer cell was found to be a mononuclear cell that had presumably undergone stimulation since it not only killed NK sensitive K-562 cells, but also NK resistant Daudi target cells.