Leukocyte Migration Test Research Articles

A Study on Drug-Induced Hypersensitivity Hepatic Injury by Leucocyte Migration Test

The identification of allergenic drugs was performed in 79 patients suspected of suffering from drug-induced hepatic injury using the leucocyte migration test (LMT). Moreover, the LMT was performed on the administered drugs in 35 patients who had been receiving drugs but had not yet manifested any allergic symptoms (patients without hypersensitivity to drugs). If either leucocyte migration activating factor (LMAF) or leucocyte migration inhibitory factor (LMIF) was detected in the LMT either with or without the patient's serum, then the test results were regarded as positive. The proportion of LMT positives was 67.1% in 79 patients suspected of suffering drug-induced hepatic injury and 8.6% in 35 patients without any such hypersensitivity to the drugs, and therefore the positive rate was significantly higher in the patients suspected of suffering drug-induced hepatic injury than in the patients without hypersensitivity to the drugs (p<0.0001, X2-test). The proportion of LMT positives was 45.6% for tests with the patient's serum and 46.8% for tests without the patient' s serum, and there was no substantial difference in the LMT-positive rate between both tests. In addition, no substantial difference was observed among the four clinical types of hepatic injuries. In addition, LMAF was detected in 34.2% and LMIF in 38.0%, and no substantial difference was seen in the rate of detection between both factors. However, concomitant symptoms were observed in 34 cases (43.0%). The proportion of LMT positives was 82.4% in the cases with concomitant symptoms and 55.6% in those without concomitant symptoms. Accordingly, the proportion of LMT positives were significantly higher in the cases with concomitant symptoms than in those without concomitant symptoms (p<0.05, X2-test). LMT-positive drugs were detected in 57 cases, in which 38.6% were β-lactam antibiotics and 17.5% were non-steroidal antiinflammatory drugs. LMAF was more frequently detected in patients with β-lactam antibiotic-induced hepatic injury (p<0.01, X2-test), while LMIF was detected more frequently in the patients with non-steroidal antiinflammatory drug-induced hepatic injury (p<0.01, X2-test). The latent period from the start of drug administration to the onset of hepatic injury was 6.3 days in the patients with β-lactam antibiotic-induced hepatic injury but 33.7 days in those with hepatic injury induced by central nervous system drugs, except for non-steroidal antiinflammatory drugs (p<0.05, t-test).Our findings indicate that the LMT is a valuable method for identifying allergenic drugs in drug-induced hypersensitivity hepatic injury, while cell-mediated immunity was shown to be closely involved in the pathogenesis of hepatic injury with concomitant symptoms and the patient's serum may thus play little role in the pathogenesis of drug-induced hepatic injury. Furthermore, the pathogenic mechanism of β-lactam antibiotic-induced hepatic injury was also found to possibly be different from that of non-steroidal antiinflammatory drug-induced hepatic injury. The latent period from the start of drug administration up until the onset of β-lactam antibiotic-induced hepatic injury may be shorter than that of hepatic injury induced by central nervous system drugs, except for non-steroidal antiinflammatory drugs.

Alcoholic liver damage–toxicity, autoimmunity and allergy

Alcoholic liver damage–toxicity, autoimmunity and allergy

Cross-antigenicity between penams and cephems by intradermal skin test and leucocyte migration test in guinea pigs.

The delayed-type hypersensitivity (DTH) reactions for penams or cephems of beta-lactam antibiotics were investigated by intradermal skin test and leucocyte migration test (LMT) in guinea pigs. The animals were immunized with ampicillin (ABPC) or cephalexin (CEX) using Freund's complete adjuvant. The cross-reactivities among ABPC, penicillin G (PCG) and cloxacillin as penam and CEX, cephalothin (CET) and cephalosporin C (CEPC) as cephem and phenylglycine (PhGly), which is the amino acyl side chain of ABPC and CEX, were examined. By intradermal reaction, ABPC-sensitized animals showed a cross-reaction with CEX, PCG and CET, but CEX-sensitized animals did not cause cross-reaction with ABPC. The CEX-sensitized group exhibited slight cross-reactions to CET and PhGly. PhGly exhibited low immunogenicity only in maximization test of guinea pig. The above results indicate that there is the difference in cross-reactivity between penams and cephems in skin test. In LMT, all the ABPC-sensitized animals reacted with ABPC and showed cross-reactions with all drugs tested. The CEX-sensitized group reacted with 4 out of 7 animals with CEX and exhibited cross-reactivities to ABPC, PCG, CET, CEPC and PhGly. The cross-reactivity between intradermal skin reaction and LMT elicited some different results.

Application of leucocyte migration tests to detection of allergenic drugs in patients with hypersensitivity to β-lactam antibiotics

In 90 suspected cases of beta-lactam hypersensitivity manifesting exanthema, pyrexia and hepatopathic symptoms, the identities of the allergenic drugs were investigated by immediate type skin reactions, sensitized erythrocyte agglutination tests and leucocyte migration inhibition tests (LMIT). In addition 30 control patients were tested for hypersensitivity to ampicillin, cephalexin and latamoxef. The control patients manifested the same symptoms as the patients with suspected hypersensitivity to beta-lactam but had not been treated with beta-lactam antibiotics. In the immediate type skin reaction, all the control cases as well as all the suspected beta-lactam hypersensitive patients displayed negative reactions to each of the antibiotics tested. In the sensitized erythrocyte agglutination tests, no antibodies to any of the three drugs were detected in the control group, while antibodies with a titre of 1:2 or more were detected in only 7% (6/90) of the suspected beta-lactam hypersensitive patients. On the other hand, in the LMIT, only 4% (4/90) of the control patients displayed positive reactions to the three test antibiotics, whereas the suspected sensitizing drugs elicited a high rate of positive responses (68/90; 76%) in the group of suspected beta-lactam hypersensitive patients. Broken down into symptomatic categories, response to the LMIT was positive in 41 of 58 cases of suspected drug rash (71%), 26 of 26 cases of suspected drug fever (96%), and 20 of 24 cases of suspected drug-induced hepatopathy (83%). Thus, the pyrexial group exhibited a particularly high ratio of positive reactions, and in fact exanthemic cases with concomitant symptoms (eosinophilia, pyrexia, hepatopathy, etc.) also displayed a notably high ratio of positives (19/20; 95%). The overall results indicated that delayed type hypersensitivity (DTH) is largely involved in exanthema, pyrexia and hepatopathy induced by beta-lactam antibiotics, and that the LMIT constitutes an effective means of detecting allergenic drugs in patients with beta-lactam hypersensitivity.

Ovine hydatidosis: some immunological characteristics of the seronegative host.

The immune status of a group of sheep naturally infected with hydatid cysts was investigated. Low specific anti-hydatid antibody (Sab) titres and negative results in intradermal and leucocyte migration tests occurred in hosts with high hydatid antigen-stimulated lymphocyte transformation (LT). Total (non-specific) IgG was elevated in infected sheep and was positively correlated with LT. Circulating hydatid antigen (cAg) was detectable in some infected sheep, but not in all of those with low Sab. Relationships between cAg, Sab and immune complex levels suggest that most cAg is present in immune complexes.

The dermal cellular infiltrate and cell-mediated immunity in skin carcinomas.

The cell-mediated immunity in nonmelanotic skin cancer was studied in 115 patients. The diagnosis and type of skin cancer was histologically evaluated. The T cell levels and leukocyte migration test in preoperative patients with squamous cell skin carcinoma (SCSC) and metatypical basal cell epithelioma were significantly lower than in the noncancer control population. The immunocompetence was not impaired in patients with basal cell epithelioma (BCE) and even in those with BCE that showed deep invasiveness and paucity of lymphocytic infiltration. There was a direct correlation between cell-mediated immunity and the lymphocytic infiltration and differentiation of the cells in SCSC.

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Blood samples from more than 1,000 patients with colorectal cancer were tested in the leukocyte migration test (LMT) and by a radioimmunoassay for carcinoembryonic antigen (CEA) for evaluation of: 1) the correlation of LMT response to the status of disease and 2) the use of LMT and CEA determination in the monitoring of patients after surgical removal of the primary tumor. A total of 320 patients, which were tested repeatedly, constituted the key group of the investigation: Of these, 56 patients developed recurrences or metastases whereas 217 patients showed no sign of reactivated disease. The group of patients with persistent absence of disease showed test profiles of either persistent negativity (50%) or conversion of one or both parameters; persistently positive profiles were rarely encountered. Patients with recurrences and metastases showed either positive profiles or conversion of positivity; only 3 patients were negative in both tests. However, despite significant disease-related response of the LMT, the test is not recommended as an adjunct to postsurgical monitoring, because of its technical complexity.

Cell-mediated and humoral alloimmunreactions after subperiostal implantation of allogenic demineralized dentin in humans

13 patients suffering from local atrophy or loss of the alveolar process of the jaws were treated by subperiosteal implantation of allogenous, decalcified, sterilized dentin with the purpose of augmentation of the alveolar process. All patients, including 9 donors of dentin were ABO-, Rh-, and HLA tissue typed. Antidonor cell-mediated immunity (CMI) and humoral-mediated immunity (HMI) of the patients were monitored during the first 6 postoperative months. The CMI and HMI were assessed by means of the leucocyte migration test (LMT) and the complement dependent microlymphocytotoxicity test (CDQ with donor leucocytes as antigen and target cells, respectively. Compared to 66 normal non-transplanted controls, the leucocyte migration inhibition against desintegrated donor leucocytes as antigen was significantly increased for 2 of the recipients during the control period. 1 of these recipients showed a simultaneous HMI reaction 6 weeks after implantation. The remaining CMI and HMI values of the recipients were within normal range. These results were not correlated with the histocompatibility match grades of the donor recipient pairs. The present investigation supports the suggestion that implantation of allogenous, decalcified and sterilized dentin occasionally is capable of eliciting a measureable alloimmune response.

Immunology of dermatophytosis. Experimental study.

By causing experimental lesions of dermatophytosis in rabbits inoculated and reinoculated with T. mentagrophytes var. granulosum and T. rubrum a study was carried out of the state of cellular immunity response, during infection, and also of the antigens responsible for the sensitization. Cellular immunity response was detected using the leucocyte migration test (L.M.T.) in the presence of antigenic compounds of the 'Keratinase' of Eleuterio et al.

Cell-mediated immunity to human oral carcinoma, and cross reactivity to patients with oral leukoplakia demonstrated by the leukocyte migration test.

Tissue extracts from 18 oral carcinomas were tested in the leukocyte migration capillary technique (LMCT) against autologous leukocytes and against leukocytes of healthy matched controls. The normal range was defined as the mean migration index (MI) of the leukocytes of the normal controls +/- 2SD. Three carcinoma patients showed significant inhibition and one was on the lower borderline of the normal range. None showed stimulation. When leukocytes of leukoplakia patients were tested against normal intestine and two homologous oral carcinoma extracts, significantly different MI's were found between carcinoma and normal intestine. The findings are compatible with a state of tissue-specific, cellular hypersensitivity against antigenic components present in oral leukoplakia and in oral carcinoma, and is possibly common between the two.

Macrophage migration stimulation factor.

Macrophage migration stimulation factor.

Leukocyte migration test (LMT) in patients with thyroid disease: the response to human thyroid subcellular fractions.

The response of circulating leukocytes to thyroid subcellular fractions was investigated in 19 patients with Graves' disease, 15 patients with Hashimoto's thyroiditis, 7 patients with toxic adenoma, 19 patients with nontoxic goiter and in 10 healthy students as control subjects. For this purpose, the leukocyte migration test of Soborg and Bendixen was performed against human crude thyroid extract (CTE), cell plasma membranes, nuclei, ribosomes, mitochondria and microsomes. Our results show positive LMT against: 1) CTE in patients with Graves' disease (61 +/- 13, p less than 0.001) and Hashimoto's thyroiditis (65 +/- 11, p less than 0.001) compared to controls (90 +/- 11); 2) cell plasma membranes in patients with Graves' disease (41 +/- 14, p less than 0.001) and Hashimoto's thyroiditis (64 +/- 21, p less than 0.05) compared to controls (88 +/- 19); 3) nuclei in patients with Graves' disease (53 +/- 25, p less than 0.001) and Hashimoto's thyroiditis (53 +/- 23, p less than 0.001) compared to controls (83 +/- 11). Our findings of circulating leukocytes sensitized to cell plasma membranes and nuclear fraction in patients with Graves' disease and Hashimoto's thyroiditis provide the additional information that these patients have a specific defect in immune-surveillance.

Effects of seminal constituents in leucocyte migration tests on infertile couples

Leucocyte migration tests were performed with cells from both partners of infertile couples in the presence of seminal constituents from the male partners. Leucocytes from fertile women were used as controls. Homogenized spermatozoa inhibited leucocyte migration in 36.1% of women and 20.7% of men from infertile couples and in 18.4% of control women. In all groups, occasional stimulation of leucocyte migration was observed (8.3%, 3.4% and 2.6% respectively). Although both types of changes in migration when considered together were statistically more frequent in the group of infertile women it was not possible to distinguish, on the basis of this test, between fertile and supposedly infertile individual women. It is suggested, however, that an inhibitory effect induced by spermatozoa on the men's own leucocytes may be of diagnostic significance. In contrast to spermatozoa, seminal plasma caused inhibition of leucocyte migration in the majority of cases — 88% of women and 71.9% of men from infertile couples and 77.8% of fertile women. Those patients whose semen lacked this inhibitory property are deserving of further study.

Production d'extraits dialysables leucocytaires chez les bovins—I. Protocole et controles de l'immunisation des animaux

Seven cows, one of which is negative control, have been immunized by a Measles' antigen and/or a brucella antigen. All of them have presented a specific immunization answer against one or both antigens used for their immunization and against those single antigens. The immunization control has been effected by some serological methods and by the inhibition of leukocyte migration test. The best immunization has been obtained by the injection of live Measles virus, Schwartz strain alone, or by the Brucella abortus B19 strain alone, when the administration of both strains showed a less intense immunization. In return, the simultaneous administration of the inactivated Schwartz strain and the live B19 strain has permitted the correct immunization of the animals, but only at the time of their second reimmunization.

Immune responses to liver membrane antigens in patients with cystic fibrosis and liver disease.

Biliary obstruction by viscid mucus, although important, may not be the only factor for the development of liver disease in some patients with cystic fibrosis. In the present study the relationship between immune responses to liver antigens and the presence of liver damage was investigated using the leucocyte migration test and lymphocyte cytotoxicity to isolated rabbit hepatocytes. Inhibition of leucocyte migration by purified liver-specific lipoprotein, derived from hepatocyte plasma membrane, was found in 9 of 11 children with liver disease, but in only 5 of 14 with cystic fibrosis and no overt liver disease (P < 0.025). Lymphocyte toxicity to isolated rabbit hepatocytes was significantly increased in 10 of 13 children with liver disease, but in only 6 of 29 children without liver disease (P < 0.001). Experiments using lymphocyte subpopulations showed that the cytotoxicity was mediated by a non-T-cell population and could be blocked with liver-specific lipoprotein in 7 out of 10 cases, suggesting that the reaction in these patients was specifically directed against liver-specific lipoprotein. The study suggests that sensitisation against liver membrane antigens, whether arising primarily or secondarily in some way to other hepatic lesions, may contribute to the progression of liver damage in cystic fibrosis.

Leukocyte migration test in subacute thyroiditis: hypothetical role of cell-mediated immunity.

There has been a paucity of data employing the leukocyte migration test (LMT) in regard to the role of cellmediated immunity (CMI) in subacute thyroiditis (ST). This has induced us toinvestigate the presence of CMI against subcellular components of human thyroid cells in patients with ST. These subcellular components obtained by the ultracentrifugation method in sucrose discontinuous gradient are: 1) cell membranes, 2) microsomes, 3) membrane-bound ribosomes, 4) mitochondria, and 5) nuclei. Moreover, a crude thyroid extract (CTE) was also used. These subcellular components have been verified by electron microscopy. The LMT technique of Soborg and Bendixen was performed in 22 patients manifesting positive thyroid autoantibody titers in the antithyroglobulin and/or antimicrosomal procedures in the course of their disorder as well as in 10 euthyroid students (volunteers). Different antigenic fractions were tested at the following concentrations [expressed as protein concentration (micrograms per ml) in 1 ml tissue culture medium] 1) membranes, 35 and 140; 2) microsomes, 85; 3) membrane-bound ribosomes, 125; 4) mitochondria, 100; 5) nuclei, 50 and 250; and 6) CTE, 40 and 80. Inhibition in the LMT test in patients with ST was observed using CTE [69 α 13 vs. 90 α 15 (controls); P < 0.001], nuclei (67 α 11 vs. 83 α 11; P < 0.001), microsomes (47 α 9 vs. 89 α 10; P < 0.001), mitochondria (60 α 6 vs. 78 α 7; P < 0.001), and ribosomes (52 α 13 vs. 85 α 20; P < 0.05). No difference from controls was in evidence when the thyroid cell membranes were usedas antigens (80 α 19 vs. 88 α 19; P = NS). The detection of CMI against nuclear, ribosomal, mitochondrial, microsomal,and crude thyroid antigens may well relate to a release of these fractions due to cytolysis brought about by viral infection and destruction.

Cross-reactivity of lung cancer patients' leucocytes in the leucocyte migration inhibition test

Panels of 3 M KCl extracts of squamous-cell carcinomas, adenocarcinomas and oat-cell carcinomas of the lung were used for a comprehensive analysis of cross-reactivity in the leucocyte migration test. Lung cancer patients' leucocytes showed positive reactivity in 69%–100% of cases (n=353). No significant differences were observed when data were grouped with respect to the histological type of the tumours used for extraction or of the tumours of the leukocyte donors. Leukocytes of patients bearing tumours of nonpulmonary origin exposed to lung cancer extract panels and leukocytes of lung cancer patients exposed to gastrointestinal cancer extract panels were definitely less reactive (35%–47% and 6%–38%, respectively). However, a high reaction frequency was found in patients with lung metastases from different nonpulmonary tumours. This group of patients also frequently showed reactivity (52%) with normal lung tissue extracts. Patients with benign lung diseases reacted positively with lung tumour extracts in 25%–39% of cases, but donors with other benign disease and healthy controls were virtually nonreactive (0–14%).

Tumour-associated antigens in culture medium of malignant melanoma cell strains

The presence of melanoma-associated antigens naturally shed from cultured melanoma cells in spent culture medium was investigated by means of a leukocyte migration test and culture medium from four melanoma and two control cell strains.

The effect of histamine on leukocyte migration test in man: I. Demonstration of a LIF production inhibitor (LIF-PI)

The inhibition by BCG of leukocyte migration was abolished by histamine in human donors with a positive tuberculin skin test. This effect was related to two mechanisms: a direct stimulation of polynuclear cell migration by histamine, and the production of a LIF production-inhibiting soluble factor (LIF-PI) by nylon-adherent T lymphocytes with receptors for histamine. This factor was not dialyzable, had a molecular weight below 50,000, and was absorbed on aggregated human immunoglobulins. Cells which produced this factor behaved as suppressors of the in vitro human delayed hypersensitivity reaction.

Delayed hypersensitivity to human encephalitogenic protein as assayed by agarose leucocyte migration in multiple sclerosis patients

Using a leucocyte migration test (Clausen's direct agarose gel migration method) hypersensitivity to human encephalitogenic protein has been examined in 50 multiple sclerosis patients (group 1), 50 healthy persons (group 2) and 25 patients with other neurological diseases (group 3). In group 1, 30 MS patients (60%) show an abnormal migration index, manifested either as inhibition or stimulation of migration; 29 controls in group 2 (58%), 11 O.N.D. patients in group 3 (44%) show an abnormal migration index. These results mean that lymphocyte hypersensitivity to myelin basic protein appears neither to be constant nor specific to multiple sclerosis. Three migration index curve types at different antigen concentration are obtained: monophasic curves within the normal index zones; monophasic curves staying in the inhibition or stimulation zone and biphasic curves with dose-effect relationship. Whatever the antigen used, this dose-effect relationship implies that the test must be carried out at different concentrations. The meaning of spontaneous sensitisation in healthy controls is discussed.