Group determination of 14-membered macrolide antibiotics and azithromycin using antibodies against common epitopes.

Erythromycin (ERY), clarithromycin (CLA), roxithromycin (ROX), and azithromycin (AZI) are macrolide antibiotics widely used in livestock and human medicine. Therefore, they are frequently found as pollutants in environmental water. A method based on indirect competitive enzyme-linked immunosorbent assay (ELISA) for group determination of these macrolides in foodstuffs, human biofluids, and water was developed. Carboxymethyloxime of clarithromycin (CMO-CLA) was synthesized and conjugated to bovine serum albumin (BSA) and gelatin to prepare immunogen and coating antigen with advantageous presentation of target epitopes, l-cladinose and d-desosamine, common for these analytes. Antibodies generated in rabbits were capable of recognizing ERY, CLA, and ROX as a group (100-150%), and AZI (12%) and did not cross-react with ERY degradants, which lack antibiotic activity. Assay displayed sensitivity of determination of 14-membered macrolides (IC50=0.13-0.2ng/ml) and low limit of detection (LOD) that was achieved at 0.02 to 0.03ng/ml. It allowed performing analysis of milk, muscle, eggs, bovine serum, water, human serum and urine, and avoiding matrix effect without special pretreatment using simple dilution with assay buffer. For 15-membered macrolide AZI, the corresponding characteristics were IC50=1.6ng/ml and LOD=0.14ng/ml. The recoveries of veterinary and human medicine macrolides from corresponding matrices were validated and found to be satisfactory.

Acute respiratory distress caused by erythromycin hypersensitivity.

Severe respiratory distress developed in a patient after ingestion of two tablets of erythromycin (Erythrocin) stearate. Complete atelectasis of the left lung was found. The patient was treated intravenously with 200 mg of prednisolone sodium tetrahydrophthalate. On the same day the patient's condition improved dramatically and repeated chest roentgenograms disclosed an almost complete expansion of the atelectatic lung. Using the indirect mast cell degranulation test and the inhibition of direct mast cell degranulation test, we have shown the presence of IgE and non-IgE antibodies (heat stable) against erythromycin. This suggests that an allergic reaction of type 1 and type 3 participated in the course of the clinical picture.

Erythromycin resistance in Danish Staphylococcus aureus hospital strains with emphasis on erythromycin consumption.

Erythromycin resistance was analyzed in 280,415 Danish Staphylococcus aureus strains isolated from hospitalized patients and phage-typed in the period 1967 to 1987. Multiresistant, erythromycin-resistant strains decreased during the whole period, while strains resistant only to penicillin and erythromycin increased after 1972. This increase coincided with an increase in erythromycin consumption in Denmark from 0.4 DDD/1000 persons/day in 1978 to 1.9, in 1987. Erythromycin consumption in nine large Danish hospitals was significantly associated with the level of erythromycin-resistant S. aureus at the hospitals.

Immunological and anti-inflammatory effects of clarithromycin: inhibition of interleukin 1 production of murine peritoneal macrophages.

The immunological and anti-inflammatory effects of clarithromycin (CAM), a new oral macrolide antibiotic, were examined in in vitro models such as lymphocyte transformation (LTF) of murine spleen cells, interleukin 1 (IL-1) production of murine peritoneal macrophages and IL-1-induced proliferation of C3H/HeJ mice thymocytes; the results were compared with those achieved by erythromycin (EM). CAM suppressed these responses much more than EM. Murine peritoneal macrophages precultured with CAM showed diminished IL-1 production, but macrophages precultured with EM did not, indicating that CAM has suppressive effects on the early phase of IL-1 production of murine peritoneal macrophages. Suppressive effects of CAM on IL-1 production by macrophages and proliferation of lymphocytes were independent of prostaglandin biosynthesis, since this drug had no effect on cyclooxygenase activity. Additional immunosuppressive and anti-inflammatory activities of CAM may explain its superior clinical effect.

[Immunologic aspects of macrolide antibiotics]. / Imunologické aspekty nekterých makrolidových antibiotik.

The authors investigated in an experimental and clinical study the part played by the macrolide antibiotic, tylosine, in the elevated immunological reactivity of the present population to Erythromycin. The first assembled results justify the statement that tylosine used in veterinary practice (sometimes without control) may indice acrossed immunological reactivity against Erythromycin. The authors discuss also other undesirable manifestations after Erythromycin administration. The assembled data confirm the need of an aimed and controllable administration of antimicrobial preparations in human as well as in veterinary medicine.

[Immunologic reactivity of tylosin and erythromycin]. / K otázce imunologické reaktivity tylozinu a erytromycinu.

The authors investigated the antibody response to erythromycin and tylosine in experiments and in a clinical investigation. The results proved in 90-100% a crossed immunological reactivity of the two macrolid antimicrobial preparations. In view of the wide use of tylosine in veterinary practice and its presence in residues of slaughter animals as well as in serological tests, it is very probable that this preparation participates in a major way in the increased incidence of allergic reactions against erythromycin in humans. Clinical results confirmed this assumption.

In vitro detection of specific IgE antibodies to erythromycin.

BACKGROUND: In vitro tests for detecting drug-specific IgE would be useful in identifying patients at risk for immediate hypersensitivity reactions to therapeutic doses of a drug. OBJECTIVE: We attempted to verify that IgE-mediated reactions to erythromycin occur and to identify IgE antibodies specific for erythromycin in serum from a patient who had urticaria immediately after administration of the drug. METHODS: Skin prick testing was performed on the patient and five control subjects. Serum from the patient, pooled sera from nonatopic subjects allergic to common aeroallergens, and cord blood controls were analyzed for erythromycin-specific IgE by radioimmunoassay. Sepharose (Pharmacia, Uppsala, Sweden) was used as solid phase covalently linked to erythromycin. RESULTS: We were able to detect erythromycin-specific IgE antibodies in serum from the patient who had an allergic reaction to this antibiotic, but specific IgE could not be detected in control sera. CONCLUSION: Immunologic IgE-mediated reactions to erythromycin do occur, and in vitro diagnosis of such reactions can be made by using Sepharose as a solid phase covalently linked to this drug.

The safety of topical erythromycin.

The prevention of allergic complications depends on the use of topical medications which have proved to be rare sensitizers. This is particularly true for stasis ulcers and eczemas which are notoriously readily sensitized. Topical erythromycin has proved to be such a non-sensitizing topical antibiotic. In a series of 60 patients with stasis ulcers, erythromycin in the form of Ilotycin ointment (erythromycin in petrolatum) was applied. Not a single instance of allergic sensitization occurred. No instances of sensitization with positive patch tests to topical erythromycin have been reported. Maximization tests with topical erythromycin did not produce sensitization. Topical erythromycin, an effective antibiotic, should be substituted for neomycin which is a common sensitizer.

Incidence of antibodies to antibiotics in subjects with chronic staphylococcal diseases.

Antibody response to antibiotics in patients with chronic staphylococcal diseases was studied and the results were compared with the production of specific antibodies. Increased titres of antibodies to antibiotics were found in cases with inadequate antibody response. The possibility of an immunosuppressive effect of antibiotics on the formation of specific antistaphylococcal antibodies is discussed.

Urticaria from erythromycin.

We report an adverse cutaneous reaction (urticaria) due to erythromycin. A positive skin prick and leukocyte histamine release tests, as well as a positive single-blind, placebo controlled oral challenge to erythromycin, strongly suggest an IgE mediated hypersensitivity mechanism.

Evidence of antibodies to erythromycin in serum of a patient following an episode of acute drug-induced hepatitis.

BACKGROUND: A patient who had taken erythromycin orally developed mild signs of hepatocellular damage and appeared to became sensitized. After a fortuitous oral intake of the antibiotic, 18 months later, the patient developed an extensive intrahepatic cholestasis. OBJECTIVE: To confirm the allergic nature of this form of hepatitis by demonstrating the presence of serum antibodies directed at the drug. METHODS: Primary cultured human hepatocytes were used to investigate whether erythromycin can covalently bind to hepatocytes; and to demonstrate the presence of drug-directed immunoglobulins in the serum of the patient. RESULTS: As a consequence of its metabolism by hepatocytes, a small percentage of erythomycin remained bound to hepatocyte proteins. The extent of covalent binding varied among hepatocytes from different donors, and was not observed in metabolically inactive hepatocytes. Antibodies able to bind erythromycin were detected in the serum of the patient. When this serum was added to a human hepatocyte culture that had been previously incubated with the drug, binding of immunoglobulins was observed around the cell membrane and close to pseudo-bile canaliculi, thus indicating the presence of antibodies able to recognize erythromycin-labelled hepatocytes. CONCLUSIONS: The existence of serum antibodies directed at erythromycin haptens in this patient strongly suggests an allergic mechanism for this drug-induced hepatitis.

Lymphocyte reactivity to ex-vivo drug antigens in drug-induced hepatitis.

The diagnosis of drug-induced hepatitis is usually based on clinical criteria, with emphasis on both the temporal relationship between drug intake and liver injury and the exclusion of alternative causes. In vitro tests of lymphocyte sensitization to drugs are considered to have a low sensitivity. We investigated the possibility of detecting lymphocyte reactivity to drugs in drug-induced hepatitis by analyzing the lymphocyte proliferative responses to ex-vivo drug or metabolite antigens to improve the sensitivity of the in vitro test. Lymphocyte proliferative responses to five different concentrations of the drug and to ex-vivo drug antigens (serum collected from normal subjects after the ingestion of the drugs) were analyzed in 25 patients with a clinical diagnosis of drug-induced hepatitis, 27 healthy subjects and 10 individuals with a recent exposure to the same drugs without development of adverse drug reactions. In seven of the 25 patients, lymphocyte reactivity to drugs was detected (28%). The use of sera collected from healthy volunteers after drug intake (ex-vivo drug antigens) and the addition of a prostaglandin inhibitor to the cultures allowed the detection of lymphocyte sensitization in seven additional cases, increasing the detection ability from 28% to 56%. We suggest that the use of ex-vivo drug antigens may represent a significant contribution to the identification of the drug involved in cases of drug-induced hepatitis.

Erythromycin--three decades later.

This article reviews some of the more recent developments relating to the clinical usage of erythromycin. The bactericidal and tissue-penetrating properties of this antibiotic are described and the suggestion that erythromycin has a very useful spectrum of activity in respiratory tract infections is supported by a variety of studies. We examine the increasing application of erythromycin in an extending number of problems such as sexually acquired disorders, a variety of diseases produced by infection with Legionella species and the enteritides associted with Campylobacter and Shigella species. The influence of erythromycin in particular, and of antimicrobial agents in general, on the immune system of the host is discussed. The immunomodulatory capacity of the antibiotics used deserves more attention. The interactions of erythromycin with theophylines and with carbamazepine are noted and amplified, and the consequences of the binding of erythromycin to plasma alpha 1-glycoprotein are examined. After some 3 decades of use, this remarkably safe antibiotic continues to display activities which deserve the attention of the clinician.

[Effect of penicillin and erythromycin on immunity in angina]. / Vliianie penitsillina i éritromitsina na immunitet pri angine.

The number of T- and B-lymphocytes in the blood of patients with quinsy was practically the same with the use of either penicillin (101 persons) or erythromycin (115 persons). The immunoglobulin concentration in the patients treated with penicillin at the beginning of the disease was normal, while during the period of reconvalescence the concentration of IgA and IgM increased. The concentration of IgA in the patients treated with erythromycin decreased at the beginning of the disease, especially when the disease was repeated. During the reconvalescence period it came to normal, while the level of IgM increased. When the patients were treated with penicillin, the increase in the titers of antistreptolysine-0- and antistreptokinase in the course of the disease was observed in 17.5 and 33.3 per cent of the patients, respectively. When the patients were treated with erythromycin the respective figures were 29.1 and 53.0 per cent. The shifts in the count of blood neutrophils including the functionally active ones in both groups of the patients were the same. It is concluded that penicillin prevented formation of humoral immunity in patients with quinsy because of its rapid antibacterial effect eliminating stimulation of the host immunocompetent cells. Erythromycin in addition inhibited production of IgA. The antibiotics had no effect on the number of T- and B-lymphocytes and neutrophils in the blood and function of T-lymphocytes and microphages.

Adjuvant effect of a 14-member macrolide antibiotic on DNA vaccine.

Macrolide antibiotics have unique immunomodulatory actions apart from their antimicrobial properties. We examined the effect of erythromycin (EM), a 14-member macrolide, on the immune response to a DNA vaccine that induces a T-helper-1 (Th1)-biased immune response through a Th1-promoting adjuvant effect of unmethylated CpG motifs within plasmid DNA. EM enhanced Th1 responses in plasmid DNA-immunized mice as measured by antigen-specific IgG2a antibody production, interferon-gamma production by antigen-specific CD4(+) T cells, and cytotoxic T lymphocyte responses. EM augmented the accessory cell activity of unmethylated CpG DNA-stimulated antigen-presenting cells (APCs), suggesting that EM enhances Th1 responses to a DNA vaccine, possibly through augmentation of accessory cell activity of APCs stimulated with CpG motifs within plasmid DNA.