Cutaneous Basophil Hypersensitivity Reaction Research Articles

Effect of supplementation of ashwagandha and vitamin E on haematological and immunological parameters of vencobb-400 broilers

Indian poultry industry is fast growing and poultry meat is not subjected to cultural and religious restrictions. Utilization of medicinal plants as feed ingredient for broilers is increasing due to its growth promoting quality and capacity to reduce antimicrobial resistance among broilers. Vitamin E enhances the immune competence of chickens and restores impaired immunity. The present experimental study was undertaken to study the effects of vitamin E and Ashwagandha (Withania somnifera) on haematological and immunological parameters of VenCobb-400 broilers. Ashwagandha and Vitamin E were supplemented in broiler chicken feed at different levels for different group. The haematological parameters viz. haemoglobin concentration, packed cell volume, total erythrocyte count, total leucocyte count and differential leucocyte count were assessed as per standard procedures. The cell mediated immune response was estimated by skin hypersensitivity test using 1% 2, 4-dinitrochlorobenzene reagent in acetone and it was evaluated after 24, 48 and 72 hours post inoculation. The data obtained in the present study were analyzed statistically. The broiler groups treated with both Ashwagandha and Vitamin E simultaneously, showed significantly higher values of haematological parameters when compared to that of other groups. The cell mediated immunity of birds analysed by cutaneous basophil hypersensitivity reaction was found at higher levels in Ashwagandha and Vitamin E treated groups. Supplementation of 2.5 g Ashwagandha and 1 g Vitamin E per Kg of feed was found to be optimum combination for better haematological and immunological attributes for broilers. This may be due to the synergistic effect of Ashwagandha and Vitamin E on the haematology of broilers.

Effects of black cumin seed (Nigella sativa L.), a probiotic, a prebiotic and a synbiotic on growth performance, immune response and blood characteristics of male broilers

This study was performed to investigate and compare the effect of different feed additives on growth performance, slaughter parameters, immune response, leukocyte profile and blood biochemical parameters of broilers. A total of 350 day-old male broiler chickens (Ross 308) were randomly assigned to seven treatments with five replicates each (10 chickens per replicate). Chickens were fed either a corn-soybean meal basal diet (control) or the basal diet supplemented with 5g/kg black seed (BS1), 10g/kg black seed (BS2), 20g/kg black seed (BS3), 1g/kg probiotic (Pro), 1g/kg prebiotic (Pre) or 1g/kg synbiotic (Syn). Average body weight gain (BWG), feed intake (FI) and feed conversion ratio (FCR) of broilers were determined during the starter (0 to 10 days), grower (11 to 28 days) and finisher (29–42 days) periods, and also for the total experimental period (0–42 days). Three chickens per replicate were randomly selected to evaluate a cutaneous basophil hypersensitivity reaction to phytohaemagglutinin-P (PHA-P) on days 35 and 36. Blood samples (2 samples per replicate) were taken for measuring antibody responses (days 28 and 42), and also for hematological and biochemical analysis (day 42). After 6 weeks, BWG was higher for Syn-fed chickens compared with control group (P<0.05). From 0 to 42 days, FCR was improved by diets BS2, Pro and Syn by approximately 6.3, 6.3 and 7.3% respectively, compared to that of the control group (P<0.05). The toe web swelling, as cell mediated response to PHA-P, in the chickens fed diets BS3 and Syn was respectively 30.5 and 27.1% greater than in chickens fed control diet (P<0.05). All diets, except diets BS1 and BS2, increased primary antibody response against sheep red blood cell (SRBC) at 28 days (P<0.05). Moreover, broilers fed diets BS3, Pro and Syn showed the higher secondary anti-SRBC titers and lower plasma triglyceride, cholesterol and low-density lipoprotein cholesterol levels at 42 days (P<0.05). Other measured parameters, including FI, slaughter parameters, leukocyte profile, heterophil to lymphocyte ratio and plasma glucose and high-density lipoprotein cholesterol levels were not affected by the treatments (P>0.05). In conclusion, dietary supplemented with probiotic, synbiotic or 10 to 20g/kg black seed improved FCR of broilers and improved plasma lipid profile and antibody-mediated immunity. These findings justify further investigations to evaluate the efficacy of these additives before field application.

Dietary arginine stimulates humoral and cell-mediated immunity in chickens vaccinated and challenged against hydropericardium syndrome virus

The effects of dietary supplement of arginine on protective humoral and cell-mediated immune responses of broiler chicks vaccinated and challenged against hydropericardium syndrome virus (HPSV) were investigated and compared with those of 2 reference drugs (cyclophosphamide and cyclosporine). Percentage ratios of lymphoid organs (bursa, spleen, and thymus) to BW, postvaccination and challenge serum antibody responses to HPSV, cutaneous basophil hypersensitivity reaction, peripheral lymphoproliferation, postchallenge detection of HPSV in the tissues of infected birds, and ability of chicks to resist virulent HPSV challenge were the parameters utilized to determine the effects of arginine on protective immune responses of chicks. A total of 600 chicks were used in this study. Arginine-supplemented chicks showed significant (P < 0.05) stimulation of lymphoproliferation and cutaneous basophil hypersensitivity reactions compared with untreated control chicks. Similarly, significantly higher body and lymphoid organ weights were (P < 0.05) recorded in arginine-supplemented chicks compared with untreated control chicks. The highest survival rate was recorded in arginine-supplemented HPS-vaccinated chicks compared with immune-suppressed (cyclophosphamide- and cyclosporine-treated and HPS-vaccinated chicks) and untreated unvaccinated control chicks after virulent HPSV challenges. Postchallenge tissue samples from arginine-supplemented and HPS-vaccinated chicks yielded negligible HPSV detections by virus isolation in cell culture or PCR method, or both, compared with untreated control chicks. Thus, it was concluded that dietary supplementation of arginine had beneficial effects on humoral and cell-mediated immune responses of broiler chicks against HPSV.

Cutaneous basophil hypersensitivity reaction to phytohemagglutinin in repeat breeder cows

In this study the relationship was explored between general immune reactivity, as estimated by the cutaneous basophil hypersensitivity reaction (CBHR) to phytohemagglutinin (PHA), and the level of anti-sperm antibodies of the Ig A and Ig G class in the cervical mucus and sera of cows with different reproductive results. The hypersensitivity test was performed on the day of artificial insemination when cervical mucus and sera samples were also collected. The animals were divided into groups according to the total number of inseminations, number of inseminations per calving and their age. The titer of antibodies was determined by the indirect immunofluorescence method using sperm cells suspended in Tris egg-yolk extender. Our results indicate that the intensity of CBHR is lowest in cows inseminated once or twice and in younger animals, while, there were no differences between the groups when they were formed according to the number of inseminations per calving. Titers of antisperm antibodies (ASA) of the Ig A class in cervical mucus (CM) increased with the number of artificial inseminations, number of inseminations per calving and age of the cows.

Melatonin enhances cellular and humoral immune responses in the Japanese quail ( Coturnix coturnix japonica) via an opiatergic mechanism

It is known that melatonin has important immunomodulatory properties in the Japanese quail. However, the mechanism of melatonin action on the immune system is not clearly understood in avian species. In mammals, the immunostimulatory properties of melatonin are mediated by the release of opioid peptides from activated T-lymphocytes. The present study was performed to determine if these same melatonin-induced opioids (MIO) are involved with the immunoenhancing effects of melatonin in quail. Three treatment groups were given melatonin (50 μg/ml) in the drinking water ad libitum along with naltrexone, a known opioid receptor-blocking agent. Melatonin was administered throughout the 3 week study and each bird received a daily intramuscular injection of naltrexone at a dose of 0.1, 1.0, or 10.0 mg/kg. In addition, three control groups were established that received only melatonin, naltrexone, or diluent. Evaluation of the cellular and humoral immune responses was initiated after 2 weeks of treatments. A cutaneous basophil hypersensitivity reaction to phytohemagglutinin (PHA-P) was measured to evaluate the cellular immune response. To evaluate the humoral immune response, primary antibody titers were determined 7 days post-intravenous injection with a Chukar red blood cell (CRBC) suspension. Both the cellular and humoral immune responses were significantly increased by 22 and 34%, respectively, upon melatonin exposure as compared to quail receiving diluent only. Concomitant administration of naltrexone and melatonin significantly reduced the immunoenhancing effect of melatonin across all naltrexone doses. We conclude that melatonin enhances a cellular and humoral immune response in Japanese quail via an opiatergic mechanism.

Effect of melatonin supplementation on the ontogeny of immunity in the Large White turkey poult

An experiment was performed to determine the effect of melatonin supplementation on the ontogeny of immunity in the Large White turkey poult. Melatonin (50 microg/mL) or diluent only (control) was administered via the drinking water from hatch to 28 d of age. The cutaneous basophil hypersensitivity (CBH) reaction to phytohemagglutinin (PHA-P) and primary antibody responses to Chukar red blood cells (CRBC) were determined at five intervals; 0, 1, 7, 14, and 21 d posthatch. At each of these times, including 28 d posthatch, 10 poults were killed and lymphoid organ and body weights were determined. Melatonin administration accelerated (P < or = 0.05) the development of cell-mediated (PHA-P-induced CBH reaction) and humoral (CRBC-induced antibody titer) immune responses, and these responses were elevated above controls through 21 d posthatch. The bursal weight, but not thymus or spleen, was elevated in melatonin treated poults as compared to controls. Body weights were increased in melatonin treated poults as compared to controls. These data suggest that posthatch melatonin supplementation is beneficial to neonatal immune parameters and growth responses of Large White turkey poults.

Melatonin can produce immunoenhancement in Japanese quail ( Coturnix coturnix japonica) without prior immunosuppression

In recent years, it has been determined that melatonin has important immunostimulatory properties in mammalian and avian species. Typically, this immunoenhancement has only been examined in immunosuppressed animals. The effect of melatonin on normal (unsuppressed) immune systems is yet to be evaluated in avian species. An experiment was performed to determine if transient and/or continuous melatonin treatments could enhance immune functions in Japanese quail without prior immunosuppression. All quail were kept on a short photoperiod (8:16LD) throughout the study. In this experiment, 50.0 μg/ ml melatonin was provided ad libitum to adult Japanese quail in the drinking water either continuously or for 3 h per day. Control birds received diluent continuously throughout the experiment. Both the cellular and humoral immune responses were determined immediately after 3 weeks treatment. A cutaneous basophil hypersensitivity reaction to phytohemagglutinin (PHA-P) was measured to evaluate the cellular immune response. To evaluate the humoral immune response, primary antibody titers were calculated 7 days post-intravenous injection with a Chukar red blood cell (CRBC) suspension. The cellular and humoral immune responses were significantly elevated in the transient (3 h) and continuous (24 h) melatonin treatment groups as compared to the control group (0 h). As compared to the control group, the cellular immune response was increased 25% and 38% for the 3 and 24 h melatonin treatments, respectively. The humoral immune response was increased 26% and 32% for the 3 and 24 h melatonin treatments, respectively. Furthermore, continuous (24 h) melatonin availability significantly increased the cellular, but not humoral immune responses as compared to the transient (3 h) group, given melatonin for 3 h prior to the scotophase (13:00–16:00 h). From these data, it was clear that transient and continuous administration of melatonin increased the cellular and humoral immune responses of Japanese quail without prior immunosuppression. These data suggest that the immunoenhancing effect of melatonin is not limited to reconstitution of weakened immune systems, but can be observed in normal, immunologically unsuppressed birds.

Pineal melatonin secretion, but not ocular melatonin secretion, is sufficient to maintain normal immune responses in Japanese quail ( Coturnix coturnix japonica)

Reports that plasma melatonin is an important immune regulator in avian species have been rather sparse and contradictory. Also, the primary source of immune-modulating melatonin has yet to be determined in birds. In Japanese quail ( Coturnix coturnix japonica), the pineal gland and eyes contribute roughly two thirds and one third of the melatonin found in the blood, respectively. Two experiments were conducted to evaluate melatonin as an immune modulator in Japanese quail and to determine the primary source of immune-modulating melatonin in this species. Experiment 1 was designed to evaluate the involvement of the pineal gland and the eyes in immunocompetence. Each of three groups of quail was assigned a surgical treatment and the cellular and humoral immune responses were determined 8 weeks following surgery. The surgical treatments were pinealectomy (Px), sham pinealectomy (SH-Px), and ocular enucleation (eye removal (Ex)). Experiment 2 utilized exogenous melatonin as a replacement to reconstitute immune responses in surgically immunocompromised birds. In this experiment, 50.0 μg/ml of melatonin, or diluent only, was provided to Px and SH-Px birds in the drinking water ad libitum. The cellular and humoral immune responses were determined after 8 weeks of melatonin treatment. In both experiments, a cutaneous basophil hypersensitivity reaction to phytohemagglutinin was measured to evaluate the cellular immune response. To evaluate the humoral immune response, primary antibody titers were determined 7 days postintravenous injection with a Chukar red blood cell suspension. Flow cytometric analysis of peripheral blood lymphocytes was performed to determine the relative percentage of CD4 + and CD8 + T- and B-lymphocytes in all treatments of Experiment 2. In Experiment 1, both the SH-Px and Ex surgical treatments produced similar cellular and humoral immune responses, and these responses were significantly greater than those in Px-treated birds. Pinealectomy significantly reduced the cellular and humoral immune responses from SH-Px by 25.8% and 41.3%, respectively. In Experiment 2, Px again resulted in depressed cellular and humoral immune responses. In addition, Px significantly reduced CD8 + T-lymphocyte numbers compared to SH-Px, while B-lymphocytes remained unchanged. Melatonin administration to Px birds increased the cellular (32.9%) and humoral (30.6%) immune responses to the level of control (SH-Px) birds, although this reconstitution was not due to increased CD8 + T- or B-lymphocytes. From these data, it was clear that removal of the pineal gland, but not the eyes, reduced cellular and humoral immune responses, which were reconstituted to normal levels by exogenous melatonin. These data suggest that immunodepression is only observed in birds with two thirds of the plasma melatonin removed by pinealectomy. Removal of one third of the plasma melatonin (by ocular enucleation) is not sufficient to reduce cellular and humoral responses in the Japanese quail.

Effects of Lighting Conditions and Melatonin Supplementation on the Cellular and Humoral Immune Responses in Japanese Quail Coturnix coturnix japonica

Two experiments were conducted to determine the effects of lighting conditions and melatonin supplementation on the cellular and humoral immune responses in Japanese quail. The first experiment was designed to evaluate differing light regimes as immune modulators in both adult and juvenile quail. The cellular and humoral immune responses were determined for three lighting conditions; short days (8:16LD), long days (16:8LD), and constant light (LL). In the second experiment, melatonin was administered in varying doses to adult quail placed in LL. The doses used in this experiment were 0.0, 0.5, 5.0, and 50.0 μg/ml melatonin given in the drinking water for 16 h per day for 2 weeks. The cellular and humoral immune responses were evaluated after 1 week of melatonin treatment. In both experiments, a cutaneous basophil hypersensitivity reaction to phytohemagglutinin (PHA-P) was measured to evaluate the cellular immune response. To evaluate the humoral immune response, primary antibody titers were calculated 7 days postintravenous injection with a Chukar red blood cell suspension. In the adult birds of experiment 1, both the 8:16LD and 16:8LD treatments produced similar cellular and humoral immune responses but these responses were significantly greater than those observed in LL. The juvenile birds held under 8:16LD also had significantly greater cellular and humoral immune responses as compared to juvenile birds held in LL. In experiment 2, there was a clear melatonin dose response on immune function in LL. The humoral immune response increased to a peak at the 5.0 μg/ml dose while the cellular immune response increased across all dose levels. From the present study it was clear that quail placed in daily light-dark cycles (LD), possessing a diurnal rhythm of melatonin, had significantly elevated immune responses as compared to those birds in LL. Furthermore, melatonin supplemented to birds exposed to LL was immuno-enhancing. This suggests that melatonin may be a mediator of the differences seen between LD and LL lighting conditions and may have important immune modulating properties.

Function and expression of adhesion molecules on human basophils

Although the human basophil is predominantly a circulating cell, it is known to participate in a variety of inflammatory responses. For example, basophils have clearly been identified in skin at sites of contact hypersensitivity, erythema multiforme, bullous pemphigoid, and cutaneous basophil hypersensitivity reactions and in the airways of subjects with allergic rhinitis and asthma?' 2 In addition, an influx of basophils accompanies the allergic late phase response that occurs after antigen challenge of the skin and airways. 3-s Thus basophils, like other granulocytes, can emigrate from the intravascular compartment into specific tissues. The observation that increased numbers of basophils accumulate at certain inflammatory sites suggests that they are activated and recruited to these locations. During this process it is necessary for these cells to interact with endothelial cells, parenchymal cells, and extracellular matrix proteins as they undergo margination, transendothelial migration, and enter the tissue space. Essentially all of the steps in this process involve the interaction of basophil cell surface adhesion molecules with specific adhesion molecule counterreceptors. Several extensive reviews have recently been written on cell adhesion molecule biology 6s and the potential role of these molecules in allergic diseases. 91x This article will focus on the expression and function of adhesion molecules on human basophils.

Immunological properties of cephalexin-induced delayed type hypersensitivity reaction in guinea pigs.

Immunological properties of delayed-type hypersensitivity (DTH) reaction induced by cephalexin (CEX) in guinea pigs were investigated. The animals were immunized with CEX using Freund's complete adjuvant. The time course of CEX-induced erythema showed some differences compared with that of classical DTH reaction. The erythema appeared at 6 h after intradermal administration of CEX, reached maximum size at 12 to 24 h and to be visible until 72 h. By enzyme-linked immunosorbent assay, anti-CEX antibody was detected in only one of 15 animals tested. Normal animals (recipients) which had received immune sera from CEX-sensitized animals (donors) showed no skin reaction to CEX. In contrast, reaction to CEX was observed in recipient animals which had received a local transfer of lymphocytes or T cells from CEX sensitized animals. In immunopharmacological study, cyclosporin A suppressed the skin reaction but cyclophosphamide did not. Administration of carrageenan, an inhibitor of macrophage function, had no effect on expression of the reaction. Post administration (1 or 15 h) of clemastine, an anti-histamine drug, did not affect the reaction. By histological examination, the infiltrating cell-types at the reaction site were mainly composed of mononuclear cells and neutrophils, but no basophils, indicating that CEX-induced DTH reaction is tuberculin-type DTH and not a cutaneous basophil hypersensitivity reaction.

Relationship between basophils and eosinophils in cutaneous basophil hypersensitivity reactions in guinea pigs and susceptibility to Trichostrongylus colubriformis infection

Guinea pigs with genetically determined resistance to primary infection with the nematode parasite Trichostrongylus colubriformis had more basophil and eosinophil leucocytes in cutaneous basophil hypersensitivity (CBH) reactions to keyhole limpet haemocyanin (KLH) than T. colubriformis susceptible guinea pigs. In outbred guinea pigs there was a significant correlation between the number of eosinophils in CBH reactions to KLH after 24 h and resistance to primary infection with T. colubriformis. These results suggest that examination of cutaneous hypersensitivity reactions to non-parasite antigens might be useful in predicting inherent resistance or susceptibility to parasite infection.

Isolation and characterization of salivary antigens from the female tick, Dermacentor andersoni.

The salivary glands of ixodid ticks are complex organs which are known to contain the antigens responsible for tick resistance in animals. We have identified a large number of proteins from salivary gland extracts (SGE), at least some of which are immunologically recognized by tick resistant animals and which are therefore presumed to be secreted salivary components. During the 6 to 10 day feeding process, a number of these antigens alter in concentration according to individual kinetics, and some of these changes correlate with the kinetics of skin test reactivity of SGE obtained at different times throughout the feeding period. By use of immunoaffinity chromatography we have isolated large quantities of many of the salivary antigens (SGA) contained in SGE, and found that they contain several esterase activities. SGA stimulates both immediate and delayed skin reactions in tick resistant guinea-pigs, and these reactions are about 200-fold more intense, per unit protein, than those elicited by SGE. The skin reactions to SGA are basophil-mediated and have many features in common with the cutaneous basophil hypersensitivity reactions of tick resistant animals. The demonstrated antigenic complexity of the glands may have profound implications for attempts to develop anti-tick vaccines, as it may eventually be found that candidate vaccines will have to incorporate more than one tick antigen in order to be effective.

Anti-T cell monoclonal antibodies in vivo. I. Inhibition of delayed hypersensitivity but not cutaneous basophil hypersensitivity reactions.

As part of a study of the therapeutic potential of anti-T cell monoclonal antibodies, we studied the biologic effects of 8BE6, a mouse anti-guinea pig (GP) pan-T cell monoclonal antibody, on blood and tissue T cells and on the prototypic T cell-mediated reactions, classic delayed hypersensitivity (DH) and cutaneous basophil hypersensitivity (CBH). 8BE6 reacts to a 68,000 m.w. protein probably homologous with human CD5 (T1) and murine Lyt-1. A single dose of 1.8 to 3.4 mg 8BE6 caused lymphopenia and greater than 90% depletion of 8BE6+ peripheral T cells 1 to 72 hr later, and a significant but lesser decrease of lymphocytes reacting with another pan-T cell monoclonal antibody (p less than 0.02 at 24 hr). Free serum 8BE6 was detected for up to 48 hr after administration. Immunoperoxidase stains of tissue revealed that lymphocytes in lymph nodes and spleen were coated with mouse immunoglobulin 1 hr after antibody treatment and displayed in situ capping. Subsequently, there was a loss of T cells in all tissues (spleen, lymph node, liver, and kidney) except the thymus, with normal 8BE6 antigen staining returning by 72 hr. Areas of induration of DH reactions to PPD were reduced in 8BE6-treated GP, compared with pretreatment reactions in the same GP or in control-treated GP (p less than 0.001 for both). The numbers of infiltrating T cells and fibronectin-receptor-positive macrophages were also reduced. In contrast, 8BE6 had no effect on CBH reactions, as judged by erythema and basophil counts in 1-micron sections, although fewer T cells were found in reaction sites. There were no differences in IgM, fibronectin, or Ia staining between 8BE6-treated GP and controls. In vivo administration of a single dose of anti-T cell monoclonal antibody results in a transient, highly specific depletion of T cell populations in peripheral blood and tissues except the thymus. This treatment inhibits DH but not CBH reactions by systemic and local depletion of T cells.

Characterization of a localized basophil hypersensitivity lesion in guinea pig conjunctiva

Basophils accumulate in response to antigen challenge in cutaneous basophil hypersensitivity (CBH) reactions. Two ocular diseases, vernal conjunctivitis and contact-lens-associated conjunctivitis, are also characterized by this histopathology. We have refined a model previously developed in guinea pig conjunctiva by precisely defining the site of antigen injection and correlating the site with the clinical and histologic changes. Guinea pigs were primed by an intradermal injection of keyhole limpet hemocyanin (KLH) in the flank and challenged (Day 6) by injection of a small bolus of KLH just under the conjunctival epithelium. Twenty-four hours later histologic examination showed a perivascular infiltrate of inflammatory cells containing large numbers of basophils. Eosinophils, neutrophils, macrophages, and mast cells were also seen. Serial sections of the reaction site showed discrete boundaries. At all sites examined in antigen-challenged tissues, there were significantly more basophils than in control-injected conjunctiva. Insertion of a sterile needle or injection of PBS or KLH into normal conjunctiva induced a significant increase in neutrophils and some macrophages. Injection of graded doses of antigen into the conjunctiva of primed animals, resulted in a dose-dependent increase in basophils up to 50 μg KLH (optimal dose).

Embryonic exposure to aflatoxin-B1: mutagenicity and influence on development and immunity.

Chick embryos were mutagenized in ovo in order to study developmentally related alterations in immune functions in survivors of this prenatal toxicant insult. In this experimental system, a single exposure of 6-day chick embryos to 0.1 microgram aflatoxin-B1 (AF-B1) in 10 microliters of acetone was employed, and the control embryos received 10 microliters of solvent alone. This dosage of AF-B1 administered to 6-day embryos was found to increase the incidence of sister chromatid exchanges in blood cells approximately fivefold above the baseline observed in solvent controls. A second sham control, where no solvent was administered, was included in some experiments. The cell cycle times in blood increased slightly during the initial exposure to AF-B1. However, a majority of the AF-B1 and acetone exposed embryos survived and hatched without incident. Losses occurred mainly in the latter part of embryogenesis. After hatching, no significant differences were observed in body weight between different treatment groups up to 26 weeks of age and no change in primary humoral immunity was detected. In contrast, two parameters of cell-mediated immunity, graft vs host (GvH), and cutaneous basophil hypersensitivity (CBH) reactions were both depressed as a result of exposure to AF-B1. The AF-B1 treatment group was significantly reduced in the GvH reaction compared with sham-treated controls. In the CBH assay, AF-B1-exposed chicks showed reduced immunity compared with acetone controls. These results suggest that long-term selective immune depression can occur following embryonic exposure to AF-B1.

IgE antibody-mediated cutaneous basophil hypersensitivity reactions in guinea pigs.

Cutaneous basophil hypersensitivity (CBH) reactions are a heterogeneous group of delayed time course basophil-rich immune responses that can be mediated in the guinea pig by T cells, B cells, or IgG1 antibody. This study examined whether guinea pig IgE antibody could also mediate CBH reactions. IgE antibody to picryl or oxazolone determinants was induced by immunizing Hartley strain guinea pigs pretreated with cyclophosphamide. Hyperimmune serum from these animals was passed through a heavy chain-specific anti-IgG1 affinity column. The presence of IgE anti-hapten antibody in the filtrate fraction was verified by passive cutaneous anaphylaxis (PCA) testing with a 7-day period of local passive sensitization and by the heat lability (56 degrees C, 4 hr) of PCA activity. This IgE-rich fraction and the IgG1 fraction eluted from the column with base (0.2 M Na2CO3, pH 11.3) were transferred i.v. to separate groups of normal guinea pigs. Both fractions mediated delayed time course reactions that contained basophils. Macroscopic and microscopic reactions mediated by the IgE-rich fraction were abolished with heat (56 degrees C, 4 hr). Thus, two antigen-specific factors in guinea pig serum can mediate delayed time course basophil-containing reactions: IgG1 and IgE antibodies. IgE-mediated CBH reactions are similar to the late-phase reaction that follows IgE-dependent wheal-and-flare reactions in humans. The finding that guinea pig IgE can mediate a late reaction that contains basophils makes this a possible model for the human late-phase response, and suggests that some forms of CBH may play a role in human allergic disease.

Sensitization of circulating basophils in guinea pig recipients of passive transfer of cutaneous basophil hypersensitivity (CBH) with immune serum: antigen-specific histamine release in vitro.

An in vitro histamine release assay was used to test the hypothesis that passive sensitization of circulating basophils is associated with the activity of immune serum that transfer the ability to elicit cutaneous basophil hypersensitivity (CBH) reactions. Systemic i.v. transfer of several types of immune sera that mediate CBH also led to passive sensitization of circulating basophils for antigen-specific release of histamine in vitro. In addition, we found that immune serum passively sensitizes basophils in vitro. Thus immune sera had three activities that are probably interconnected: sera will 1) passively transfer CBH in vivo, 2) passively sensitize basophils in vivo, and 3) passively sensitize basophils in vitro. These results suggest that passive sensitization of circulating basophils by immune serum contributes to the mechanism by which antibodies transfer the ability to elicit CBH reactions.

IgG1 antibody-dependent mediator release after passive systemic sensitization of basophils arriving at cutaneous basophil hypersensitivity reactions.

When antigen is injected into a 24-hr cutaneous basophil hypersensitivity (CBH) reaction of an actively sensitized guinea pig, local basophils degranulate and release histamine. This reaction is called cutaneous basophil anaphylaxis and may be antibody mediated. We now report passive sensitization of basophils at CBH sites by systemic transfer of anti-picryl immune serum. Keyhole limpet hemocyanin- (KLH) immunized animals were skin tested with KLH to elicit 24-hr CBH reactions at day 7. Anti-picryl serum was injected i.v. at various times. On day 7, blue dye was injected i.v., and then 24-hr CBH sites vs nearby normal skin were challenged with 0.1 microgram picryl-human serum albumin (Pic-HSA). An immediate increase in vascular permeability (blueing) was noted at normal skin sites due to systemic passive cutaneous anaphylaxis (PCA), and augmented blueing occurred at CBH sites compared with normal skin. Systemic passive sensitization of CBH sites occurred when antiserum was administered as little as 1 hr before challenge of CBH site. However, local administration of anti-picryl serum (as in a local PCA reaction) was not able to sensitize tissue basophils, whether antigen was administered locally or systemically. The serum factor that mediated cutaneous basophil anaphylaxis was heat-stable (56 degrees C X 4 hr) 7S IgG1 antibody. Electron microscopy of Pic-HSA-challenged CBH sites in animals that received IgG1 antibody showed that local basophils undergo anaphylactic degranulation by exocytosis. These studies suggest that basophils arriving at CBH reactions are sensitized for anaphylactic function by antibody that can be acquired in the circulation, but possibly not at the local site.

Dissociation between cutaneous basophil hypersensitivity and basophil sensitization.

We investigated the antigen-specific sensitization of basophils in the peripheral circulation of guinea pigs using an in vitro histamine release assay. Animals were primed for cutaneous basophil hypersensitivity (CBH) by intradermal injection of either keyhole limpet hemocyanin (KLH) or ovalbumin (OA) in saline. Basophils isolated from these animals 7 days after sensitization did not release histamine to the specific antigen in vitro. On day 9, however, basophils did release histamine when challenged with the antigen. These results were in contrast to 24-hr skin tests that showed a typical CBH profile both on day 7 and day 9. When animals were immunized via the footpad by emulsification of antigen in incomplete Freund's adjuvant, basophil sensitization (i.e., histamine release) either occurred earlier (day 7, KLH) or at lower antigen concentrations (OA). Thus, dissociation of in vivo CBH and in vitro basophil sensitization was demonstrated. The lack of correlation was only seen early after immunization and depended on the route of immunization used. An additional finding was a transient blood basophilia that also was dependent on the route of immunization, the antigen used, and the time post-sensitization. Our results are consistent with the hypothesis that at early times post-immunization, CBH is mediated by T cells and not via homocytotropic antibodies, but that at later times such antibodies may also play a role in the elicitation of CBH reactions.