Leukocyte Migration Inhibition Test Research Articles

Sensitisation to Mallory bodies (alcoholic hyalin) in alcoholic hepatitis.

Using a leucocyte migration inhibition test sensitisation to Mallory bodies (alcoholic hyalin) was found in a statistically significant 41% of 17 patients with alcoholic hepatitis. Patients with alcohol-induced fatty liver and cirrhosis did not demonstrate sensitisation. Mallory bodies are a characteristic feature of alcohol-induced liver damage, and immunological sensitisation to them might lead to liver cell death and cell progression of the hepatitis process.

In vitro assays of cell-mediated immunity in calves with persistent [formula omitted] infections do not correlate with delayed skin hypersensitivity

A long-term kinetic study of the immune response of four calves experimentally infected with a M. avium strain was made using the following tests: lymphocyte stimulation (LS), leukocyte migration inhibition (LMI), delayed skin hypersensitivity (DHS), and a radioimmunoassay (RIA) procedure for antibodies to M. avium . The cell-mediated immune (CMI) responses measured by these tests showed different courses during the infection period. The LS test showed several periods of peak values and the LMI test two peaks of responses of relatively shorter duration. At the end of the experimental period the DHS responses had decreased to insignificant levels, whereas the lymphocytes from all the calves responded in the LS test. No correlation could be detected between CMI and antibody mediated responses, and only two of the infected calves showed clear cut antibody responses measured by RIA. The results probably reflect the measurement of different aspects of the CMI response in the different test systems. The cyclical nature of the CMI measured by the LS and LMI tests may indicate the influence of regulatory mechanisms by suppressing lymphocyte sub-populations.

Influence of nutrition on postvaccinial tuberculin sensitivity

Response to BCG vaccination was studied in 261 apparently normal preschool children in a community. They were classified into different nutritional groups based on deficit in weight for age. In addition, nine children who had kwashiorkor and were admitted to the hospital were investigated. They were given 0.1 ml of BCG vaccine, and 6 months later, tuberculin sensitivity was assessed using 5 U of PPD. Blood samples were collected from 84 subjects and leukocyte migration inhibition was determined using the same antigen. After BCG vaccination, over 80% of children in the community showed positive tuberculin test, irrespective of the extent of growth retardation. There were no significant differences in the size of induration or the percentage of reactors between the various groups, indicating that the immune response to BCG vaccination is not affected by milder grades of malnutrition. However, the skin test was negative in most of the children who had had kwashiorkor. Leukocyte migration inhibition was similar in all the groups of children including those with kwashiorkor indicating that sensitisation of lymphocytes was not influenced by the nutritional status. In children with kwashiorkor, the leukocyte migration inhibition test was positive though the skin test was negative, suggesting that the former may be a better measure of assessing the response to BCG vaccination.

Phytohaemagglutinin-induced leukocyte migration inhibition and phytohaemagglutinin skin test for screening the cell-mediated immune response in children.

Phytohaemagglutinin-induced leukocyte migration inhibition was studied in 72 children and compared with phytohaemagglutinin skin testing. The methods were used for screening the cell-mediated immune response. The in vitro method, which requires only 10-15% of the number of migrating cells as compared with other migration techniques, proved to be more sensitive, but gave some false-positive results when compared with other immunological parameters. Nevertheless, application of the microdroplet leukocyte migration is advantageous, used either simultaneously with the skin test, or as a single screening method in prematures, mature newborns and young infants, where application and/or evaluation of the skin tests is difficult.

HUMORAL AND CELL‐MEDIATED IMMUNITY IN LARGE NON‐TOXIC MULTINODULAR GOITRE

Humoral and cell-mediated immunity was investigated in fourteen patients with non-toxic multinodular goitre and ten healthy controls by in vitro methods. These included determination of sheep erythrocyte and complement rosette-forming cells in the peripheral blood, immunoglobulin levels, titres of thyroglobulin and microsomal antibodies and migration inhibition test using thyroid extract and phytohemagglutinin. When compared with controls the patients showed high IgA levels and positive response to thyroid antigen in the leucocyte migration inhibition test. There was no correlation between the leucocyte migration results and the presence of auto-antibodies. These findings indicate a possible role of cell-mediated immunity in non-toxic multinodular goitre.

Immune Response in Rabbits to Surface Components of Extracellular and Intracellular Forms of Vaccinia Virus

The development of cellular as well as humoral immune response to extracellular and intracellular forms of vaccinia virus (ECV and ICV, respectively) and their surface antigens were studied in rabbits. Direct lymphocyte cytotoxicity and peripheral blood leukocyte migration inhibition tests were used to measure cell-mediated immune response, while neutralizing and hemagglutination-inhibiting antibodies were assayed for measuring humoral immune response. Direct cytotoxicity of lymphocytes from rabbits immunized with ECV or its surface proteins was demonstrable by day 7 after immunization, and by the end of week 3 it almost declined to pre-immunization levels. Inoculation with ICV or its surface proteins failed to induce lymphocyte cytotoxicity. In contrast, migration inhibition of peripheral blood leukocytes from rabbits immunized with ECV, ICV, or their surface proteins was observed with homologous antigens. However, leukocytes from rabbits immunized with ECV or its surface proteins also showed migration inhibition in the presence of ICV. Similarly, in the humoral immune response, neutralizing antibodies were produced against homologous as well as heterologous forms of virus despite immunization with purified preparations of ECV, ICV, or their surface proteins. Adsorption with purified ICV preparations abolished the neutralizing activity of these antisera against heterologous forms of virus. Hemagglutination-inhibiting antibodies, on the other hand, were produced only after immunization with ECV or its surface proteins. In addition, antibody-dependent cell-mediated cytotoxicity was employed to detect specific antibody response after immunization of rabbits with live virus, ECV, and ICV. Antisera raised against ECV or live virus supported antibody-dependent cell-mediated cytotoxicity, whereas ICV-antiserum failed to do so. The antibody activity present in the former antisera was abolished by absorption with cell membranes from vaccinia-infected cells but not with purified ICV. The data suggest that immunization with inactivated ECV seems to bring about interaction between host immune response (cellular and humoral) and virus-infected cells, which may, perhaps, be necessary for protection against pox virus infection. A similar interaction is unlikely to occur after immunization with inactivated ICV.

Leucocytes from patients with colon carcinoma or non-cancerous intestinal disease react differently to foetal colon extract in leucocyte migration inhibition test

Leucocytes from patients with colon carcinoma or non-cancerous intestinal disease react differently to foetal colon extract in leucocyte migration inhibition test

The nonspecific inhibitory effect of synovial tissue extracts on leucocyte migration in vitro.

The leucocyte migration inhibition test (LMT) has been used to search for specific antigens in rheumatoid synovial tissue. Synovial samples were collected from 20 patients with rheumatoid arthritis, from 1 patient with ankylosing spondylitis, and from 1 patient with pigmented villonodular synovitis. Inhibitory material was obtained from all 21 synovia with inflammatory disease but not from the noninflammatory synovium. The tissue extracts generally caused nonspecific migration inhibition when tested against a total of 157 pairs of rheumatoid and control leucocytes. However, occasional samples did induce migration inhibition restricted to either rheumatoid or control cells. The inhibitory factor was shown to be membrane associated and of high molecular weight (greater than 10(6) daltons). The results of this extensive study do not support the conclusions drawn from earlier reports based on smaller numbers of experiments. No evidence was obtained for the existence of specific antigenic material associated with the synovial membrane in rheumatoid arthritis.

Lack of correlation between carcinoembryonic antigen content of tumor extracts and leukocyte migration reactivity of tumor patients.

Tumor extracts varying in carcinoembryonic antigen (CEA) content (0.4--2,280 ng/mg protein) did not affect the reactivity of cancer patients' leukocytes in the leukocyte migration inhibition test (LMIT). In addition, variations in the plasma CEA levels of tumor-bearing leukocyte donors did not influence the frequency of significant LMIT reactivity.

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%).

Cell-mediated immune response in pigs persistently infected with a Mycobacterium avium strain

Following a persistent bacterial infection with a Mycobacterium avium strain in pigs, the lymphocyte stimulation (LS), leucocyte migration inhibition (LMI) and tuberculin skin reactivity responses were studied. The responses in the LS and LMI tests occurred within a period of 35 to 49 days after infection. Positive tuberculin skin responses were found 40 to 70 days after infection. No direct correlation was evident between the LS and LMI tests during the experimental period. The responses to purified protein derivatives (PPD) in the LMI test were of shorter duration than those seen in the LS test. The course of LS in pigs expressed high, lower and non-stimulatory responses to the tested PPDs. At the end of the experimental period, 173 days after infection, lymphocyte stimulatory reactivity to PPDs could be demonstrated in all pigs, whereas a positive tuberculin skin test was observed in only one animal. Lymphocyte responses to M avium PPD were generally higher than were the other tested PPDs.

T lymphoid cells in primary syphilis. Quantitative studies.

In previous studies to assess the role of cell-mediated immunity in Treponema pallidum infection investigations of delayed type skin hypersensitivity tests, lymphocyte transformation test, leucocyte migration inhibition tests, and histological studies of the reticuloendothelial system have been performed. In this study, the numbers of T (thymus-dependent ) lymphoid cells in 10 patients with primary syphilis (eight untreated) were estimated by the rosette technique. Results indicate a significant decrease in the number of T lymphoid cells in patients with primary syphilis.

Leukocyte Migration Inhibition Among Breast Cancer Patients in Response to Various Oncogenic Viruses<xref ref-type="fn" rid="FN2">2</xref>

The direct leukocyte migration inhibition (LMI) test was done with leukocytes from healthy women and from patients with primary operable breast cancer, benign breast disease, or head and neck cancer. Purified preparations of murine mammary tumor virus (MuMTV), Mason-Pfizer monkey virus (MPMV), and murine leukemia virus (MuLV) were used. For each virus, the lowest 10th percentile of the LMI responses of controls was used to discriminate positive from negative responses. Leukocytes from 46 of 94 (49%) breast cancer patients responded to MuMTV, which was significantly different from each of the other test groups: Positive reactions were observed in only 9 of 67 (13%) healthy persons, 2 of 32 (6%) patients with benign breast tumors, and 2 of 20 (10%) patients with head and neck cancer. Although leukocytes from 29% of the breast cancer patients responded to MPMV, this response did not significantly differ from that observed in healthy women (14%), in patients with benign disease (20%), or in patients with head and neck cancer (20%). The leukocytes from the breast cancer patients were not reactive to MuLV. LMI tests to both MuMTV and extracts of MCF-7 cultured breast cancer cells were done in 36 breast cancer patients and 40 healthy women simultaneously. Of the breast cancer patients, 75% responded to MuMTV and/or MCF-7 antigen as compared to 18% of the controls (P less than 0.005). These data suggest that leukocytes from breast cancer patients are presensitized to MuMTV and antigen(s) present in MCF-7 breast cancer cell line, but not to MPMV or MuLV.

Leukocyte migration inhibition test in patients with herpetic corneal disease (author's transl)

In 25 patients with primary and 24 patients with recurrent herpetic corneal disease the cell-mediated immunity to Herpes simplex virus antigen (type I) was measured using a leukocyte migration inhibition assay. After a period of one to 24 months 25 of the patients were tested again. The diagnostic possibilities offered by this method, especially in respect of control of immune-stimulating therapy, and its pitfalls are discussed.

Leukocyte migration inhibition in chickens immunized with Eimeria tenella.

Leukocyte migration inhibition tests (LIT) with chicken peritoneal cells were carried out by the direct and the indirect techniques in chickens immunized with Eimeria tenella. In LIT by the direct technique, migration of peritoneal cells was inhibited by adding antigen prepared from the 2nd-generation merozoites. The activities of leukocyte migration inhibitory factor (LIF) were detected from 1 week after a single inoculation with 50, 000 oocysts of 3 weeks after the last of 3 repeated inoculations with 50, 000 oocysts. Spleen cells obtained from immunized chickens elaborated LIF into the medium when they were cultivated with antigen. In LIT by the indirect technique, supernatant of the cultures of spleen cells from immunized chickens inhibited migration of peritoneal cells collected from normal chickens. The LIF activities detectable by the indirect technique were demonstrated for 5 weeks after the last of 3 repeated inoculations with 50, 000 oocysts.

Role of T- and B-lymphocytes in heterogeneity of cell-mediated reactions to bacterial antigens in man

Blood leukocytes from 30 patients with allergy to tuberculin and bacterial antigens were treated with antithymus (ATS) or anti-immunoglobulin serum (AIGS), after which the leukocyte migration inhibition (LMI) test was carried out with these antigens. ATS abolished LMI by tuberculin and sometimes by bacterial antigens (staphylococcal, streptococcal, etc.). AIGS frequently abolished LMI caused by bacterial antigens but not by tuberculin. In other cases treatment with any serum abolished LMI by antigens or, conversely, it was abolished only by treatment with both antisera in turn. The type of lymphocytes (T or B) determining the reaction to the same antigen in the secondary immune response differed in different patients and also differed in the same patient for different antigens. Five types of interaction between lymphocytes and antigen in the LMI test were distinguished.

Cellular hyperreactivity to placenta in toxemia of pregnancy

It is generally assumed that maternal immune response may be involved in some pathological conditions of pregnancy, such as toxemia. The activity of cell-mediated immunity to placenta was studied in women with gestosis. The leucocyte inhibition technique was employed to test reactivity of peripheral blood leucocytes from postpartum women. The results indicated that leucocyte migration was significantly inhibited in the presence of placental microsomal fraction when blood from toxemic women was used. Thus, about 87% out of 30 studied cases of toxemia showed positive results. At the same time, in only 22% of women with uncomplicated pregnancy was such a positive leucocyte migration inhibition test found. There was no correlation between the severity of the toxemic symptoms and the extent of inhibition. In further experiments the leucocyte migration inhibition test with the microsomal fraction antigens from cadaver kidneys was performed; only 1 out of 6 patients showed positive leucocyte migration inhibition. These findings may suggest that the normal status of the mother is altered towards cellular hyperreactivity to placental antigens in toxemic pregnancy.

Hyporesponsiveness to Virus Antigens in Rheumatoid Synovial and Blood Lymphocytes Using the Indirect Leucocyte Migration Inhibition Test

Mononuclear cells (MNC) from rheumatoid synovial tissue and peripheral blood were tested plasma pneumonia by the indirect leucocyte migration inhibition test. MNC from the eleven rheumatoid synovial tissues tested had deficient leucocyte inhibitory factor production against all antigens tested for, and this was also the case in the peripheral blood of seven juvenile rheumatoid arthritis patients (JRA). In the peripheral blood of eight rheumatoid arthritis (RA) patients there was also generally low reactivity. However, significant differences in migration indexes were found with rubella viral antigen and with PPD at 5 micrigram/ml when zero-hour and overnight incubations of the culture were compared. In contrast, MNC of peripheral blood of control donors had significant responses to PPD (19/19), mumps virus (7/11), rubella virus (10/19), cytomegalovirus (4/11), and herpes simplex type 1 virus (4/11) antigen after zero-hour culture, and no differences was seen after overnight incubation.

LEUCOCYTE MIGRATION INHIBITION TEST IN CADAVERIC RENAL TRANSPLANTATION

The leucocyte migration inhibition test has been studied in a series of 192 renal allograft recipients. Seventy-seven patients showed no evidence of inhibition in the early post-transplant course, but 31 of these demonstrated clinical evidence of rejection, a false-negative rate of 16%. The remaining 115 recipients all demonstrated inhibition, with 13 of these showing no clinical evidence of rejection, a false-positive rate of 6.7%. Early antirejection therapy on the basis of inhibition did not result in improved kidney survival when compared with those recipients who did not receive specific therapy until there was clinical evidence of rejection. The leucocyte migration inhibition test did not detect changes attributable to humoral factors, which probably accounts for the high false-negative rate, and has not proved to be sufficiently reliable to be of value clinically as a single test. A combination of tests designed to detect both humoral and cellular factors responsible for rejection deserves further study.

In vitro transfer of specific reactivity to cytomegalovirus and candida to cord blood leukocytes with dialyzable leukocyte extracts

The ability of two preparations of dialyzable leukocyte extracts (DLE, formerly termed “dialyzable transfer factor”) to induce specific responsiveness by cord blood leukocytes (CBL) in the direct leukocyte migration inhibition (LMI) assay in vitro was studied. One donor of DLE (DLE-A) was immune to cytomegalovirus (CMV), as judged by positive LMI test, and negative to candida as judged by skin test. The other donor (DLE-B) was immune to both CMV and candida. Several concentrations of DLE and of antigen were tested. High concentrations of DLE nonspecifically inhibited leukocyte migration, while low concentrations had no effect on antigen-induced LMI. Antigen-specific conversion was produced by intermediate concentrations of DLE. Both preparations transferred reactivity to CMV, but only DLE-B transferred reactivity to candida in vitro.