Leukocyte Migration Inhibition Test Research Articles

Influence of palmitoyl-3-catechin and heptyl-3-catechin on the leucocyte migration inhibition test carried out in the presence of PPD and hepatitis B surface antigen (HBsAg)

It has been shown that (+)-cyanidanol-3 ((+)-catechin) is able to stimulate in vitro the cell-mediated immune response specific for hepatitis B surface antigen (HBsAg), and might contribute to the elimination of hepatitis B virus (HBV) during HBV infection. In the present study, the possible action of two of its derivatives, palmitoyl-3-catechin and heptyl-3-catechin, on this type of immunity was investigated by adding the substances to leucocyte migration inhibition tests performed in presence of PPD and HBsAg, with leucocytes from individuals sensitized to these antigens. In normal individuals sensitized to PPD, the addition of palmitoyl-3-catechin and heptyl-3-catechin amplified the inhibition of migration by resp. 7.2% ( p < 0.05). In patients previously infected by HBV and sensitized to HBsAg, the maximum amplification was resp. 12.7% ( p < 0.001) and 7.6% ( p < 0.05). This effect was dose-dependent. These substances did not modify the leucocyte migration measured in the absence of antigen. Palmitoyl-3-catechin and heptyl-3-catechin therefore seem capable of amplifying the cell-mediated immune response. The effect of the two derivatives, which were selected because of their liposolubility, was more pronounced than the effect of (+)-cyanidanol-3. It is thus possible that the two new catechin-derivatives, not yet in therapeutical use, will also stimulate cell-mediated immunity to HBsAg, and that a more marked clinical effect might be expected.

Correlation between cell-mediated immunity and clinical forms of paracoccidioidomycosis

Cellular immune response to specific and non-specific stimulants was investigated, both in vivo and in vitro, in 29 healthy controls and in 53 previously untreated patients with the chronic isolated organic form (CIOF), the chronic mixed form (CMF) and the acute progressive form (APF) of paracoccidioidomycosis. The study included skin tests to Paracoccidioides brasiliensis antigen (PbAg) and phytohaemagglutinin (PHA), DNCB sensitization, determination of T lymphocytes and complement rosette-forming cells, lymphocyte transformation and leucocyte migration inhibition tests using PbAg and PHA. Patients displayed staggered cutaneous response to PHA and to PbAg, with marked decrease in intensity in the APF group. DNCB sensitization test and proliferative response of lymphocytes to PHA and PbAg were severely depressed in most of the patients. Leucocyte migration inhibition indices to PbAg were highly positive, while response to PHA was slightly decreased regardless of the clinical form. The number of T lymphocytes was reduced in most of patients and in them the number of complement-rosette forming cells was normal. The distribution of patients according to a suppression index, based in the results of the tests employed, revealed a tendency towards an increased degree of cellular immunosuppression from the least severe (CIOF) to the most severe (APF) clinical form of the disease. On the whole, the present study demonstrated a gamut of immunological reactivity in paracoccidioidomycosis.

Cellular hypersensitivity to human pancreatic B-cell clone in diabetes mellitus and its relationship to the presence of islet cell antibodies.

A leukocyte migration inhibition test on the human pancreatic B-cell clone (JHPI-1) was performed in 13 IDDM patients with islet cell cytoplasmic antibody (ICCA) and/or islet cell surface antibody (ICSA), 15 IDDM patients without ICCA or ICSA, 34 NIDDM patients and 17 healthy controls. The mean values for the migration index (M.I. %) in each group were 85.4 +/- 6.9, 89.1 +/- 10.9, 98.3 +/- 7.9 and 100.0 +/- 8.5. The M.I. values were significantly decreased in IDDM patients than in NIDDM patients and controls irrespective of whether or not there were islet cell antibodies in the patients' sera. When M.I. values less than 0.83 (Mean-2 S.D.) were taken as indicative of inhibition, the percentage of IDDM and NIDDM patients with migration inhibition were 32% and 0% respectively. And the decreased M.I. values in IDDM patients proved not to be due to non-specific migration inhibition by normal M.I. values, with the human fetal lung fibroblast cells (W 138) as antigen. Our data suggested that the lymphocytes of IDDM patients might be sensitized by pancreatic B-cell antigen(s) present in the JHPI-1 cells, which promoted leukocyte migration inhibition. No correlation between the migration indices and duration of diabetes mellitus in IDDM patients was observed (r = 0.254, Y = 84.9 + 0.49 X). LMT to JHPI-1 seems to be useful in detecting the abnormal cell-mediated immunity even in patients with longstanding IDDM.

Immunogenicity of glutaraldehyde‐treated bovine pericardial tissue xenografts in rabbits

Xenograft valves of glutaraldehyde-treated bovine pericardial tissue have been claimed to be an excellent choice for the replacement of malfunctioning human heart valves. These valves exhibit low thrombogenicity, hemodynamic function, and transvalvular gradients superior to their mechanical and tissue counterparts. However, there are differences of opinion concerning the immunogenic properties of these valvular devices. This study was conducted to evaluate the immune reactivity of such implants. Data obtained indicated that whole xenograft sections of both untreated and treated valvular tissue remain immunogenic following initial and secondary subcutaneous implantations in rabbits. Homologous and cross-reacting antibodies were detected by means of indirect hemagglutination test 30 days after initial implantation of the grafts. Presence of cellular immune response as judged by skin reaction and leukocyte migration inhibition tests indicated that glutaraldehyde treatment of tissues does not destroy cellular immunity. Data obtained also suggest that chemical modification of such implants by glutaraldehyde probably alters, but does not abolish some antigenic determinant sites on the tissue surface. Glutaraldehyde treatment of bovine pericardial tissue does not render the tissue immunologically inert.

Cell mediated immune response at varying age periods in relation to their nutritional status among preschool children given BCG at birth.

Cell Mediated Immune Response (CMIR) was investigated in 238 preschool children, 9 months to 6 years old, of varying nutritional status who had been given BCG at birth. Leucocyte migration inhibition test (LMIT) was used as an in vitro and Mantoux test as an in vivo measure of CMIR. The difference in the percentage of positive Mantoux test and area of induration was not significant between normal children and those suffering from mild to moderate undernutrition. No significant difference was found in positive LMIT between normal and undernourished group. Positive Mantoux test was elicited in fewer (38.7 to 26.5 per cent) children compared to positive LMIT (54.8 to 53.9 per cent) in the age group < 1 to 6 years. The results indicate that mild to moderate degree of malnutrition does not interfere with the elicitation of CMIR after BCG vaccination. Waning of CMIR occurred by the end of the first year. A negative Mantoux test does not necessarily mean absence of CMIR.

Demonstration of antibody and cellular immune response to brain extract in West and Lennox-Gastaut syndromes.

We investigated humoral and cellular immune response to brain tissues in 15 patients with West syndrome, in 9 patients with Lennox-Gastaut syndrome and in 20 healthy children. High levels of a precipitating antibody to a saline extract of brain tissue were detected in all patients; leucocyte migration inhibition test with the same antigen was found to be positive in most of them. The role of this autoimmune response in the pathogenesis of West and Lennox-Gastaut syndromes remains to the elucidated.

Immunological disturbance in West and Lennox-Gastaut syndromes.

Cell-mediated and humoral immunity were investigated in 18 patients with West syndrome, 12 with Lennox-Gastaut syndrome and 19 healthy controls. The study included determination of T and B peripheral blood lymphocytes, serum levels of IgG, IgA and IgM, skin sensibilization with DNCB, intracutaneous PHA, leucocyte migration inhibition test and lymphocyte blastic transformation in the presence of PHA. Cell-mediated deficiency was detected in 28 children whereas low levels of immunoglobulins were observed only in 6 children. Immunological disturbances were more prominent in children with West syndrome.

Lymphocyte Reactivity in Normal and Malignant Cell Proliferation Against a Phylogenetically Conserved Antigen in Fetal Extracts&lt;xref ref-type="fn" rid="FN2"&gt;2&lt;/xref&gt;

Lymphocyte reactivity to 3-M KCl extracts from fetuses of different species of origin was shown by the macrophage electrophoretic mobility (MEMB) and/or the leukocyte migration inhibition tests in tumor-bearing mice and humans. In chemical carcinogenesis, reactivity was detectable before tumors developed. Six weeks after sc injection of 1,000 micrograms benzo[a]pyrene [(BP) CAS: 50-32-8] into XVII/Bln mice, the MEMB test became positive. The latent period was 15 weeks after 1.0 micrograms BP, indicating a dose-response relationship of the phenomenon. Painting of mouse skin with 7,12-dimethylbenz[a]anthracene [(DMBA) CAS: 57-97-6], croton oil (CAS: 8001-28-3), or benzene (CAS: 71-43-2) had the same sensitizing effect as BP. In contrast to the strong carcinogens BP and DMBA that caused lymphocyte reactivity to persist until tumors developed, benzene and the promoter croton oil induced only a transient effect. Termination of treatment abolished reactivity within 12 weeks. Lymphocyte reactivity to fetal extract in normal cell proliferation was evident from the fact that two-thirds-hepatectomized rats and BCG-treated mice became MEMB-positive. In hepatectomized rats the effect was reversible according to the completion of liver regeneration. Lymphocytes from tumor-bearing mice reacted with mouse and human fetal extract as well as with extracts from different developmental stages of frogs and fish. Fetal extracts were assumed to contain a phylogenetically conserved antigen.

Kinetics of peripheral blood T cell numbers and functions in patients with burns.

Cell-mediated immunity (CMI) was assessed in 58 patients with burns involving 10% to 40% of the total body surface area (TBSA), and in 16 normal healthy individuals, by using two parameters, the total T-cell counts in peripheral circulation and T-cell function as indicated by the leukocyte migration inhibition (LMI) test. Three recall antigens from Candida albicans, S. typhi, and E. histolytica were used to elicit the LMI. T-cell function was found to be significantly depressed in burns involving more than 25% TBSA. Depression of T-cell function started after the seventh postburn (PB) day and continued until the sixtieth PB day.

Exploration des troubles de l'immunité au cours des péritonites postopératoires

Despite major diagnostic and therapeutic advances, postoperative peritonitis appeared to be still associated with a severe prognosis. The failure to react to delayed hypersensitivity skin tests was recently shown to identify patients at increased risk for sepsis. In an attempt to clarify the mechanisms of this anergy, cellular and humoral immunity was studied with in vitro tests in 12 patients treated for postoperative peritonitis. Complement was decreased in 33.3% of cases and normal in the others. No significant change was found in IgG and IgM titres, but IgA concentrations were increased in 80% of cases. A decrease in the total number of lymphocytes was observed in 41.7% of patients, related to the reduction in the total T lymphocyte count. Mitogen-induced lymphocyte transformation was studied with phytohaemagglutinin, concanavalin A, pokeweed-mitogen, and tuberculin purified protein derivative. Six patients had decreased or negative response to at least three mitogens; 91,7% had no response to tuberculin. The leukocyte migration inhibition test was negative in all cases. These abnormalities in cell mediated immunity may have been related to underlying diseases (severe nutrition depletion in 7 cases), to sepsis (septic shock in 10 cases), to repeated anaesthesias and surgical procedures, and even to drugs ( e.g. antibiotics). The presence of serum inhibitors may have been the cause of the anergy and further studies are required.

Cell mediated immunity in patients with subacute sclerosing panencephalitis

Cell mediated immunity was assessed on the basis of total lymphocytes (TL), total T lymphocytes (TTL) counts, delayed skin test responses and in vitro leucocyte migration inhibition test (LIF) production in 25 patients with SSPE, classified according to the clinical stages of the disease. The patients in stage I of the disease did not show any defect in cellular immunity while the patients in stage II showed decreased TL and TTL counts, more negative skin test responses to PHA, SKSD and PPD, and unresponsiveness to SKSD of LIF production. When the patient group was evaluated as a whole, only the TL counts and the skin test responses against SKSD differed from those in the controls. These results suggest that the defects in cellular immunity demonstrated in the patients with SSPE may be due to SSPE or the measles virus itself rather than to a genetic factor predisposing patients to SSPE.

Studies on effect of carcinoembryonic antigen on leucocyte migration inhibition test in patients with colorectal cancer

Leucocyte migration inhibition test (LMIT) is a useful method to detect tumor associated antigens (TAA) in cancer patients. Carcinoembryonic antigen (CEA) is one of the best tumor markers for gastrointestinal cancer, and especially for colorectal cancer patients, who show high plasma CEA level frequently. In this study, we performed LMIT in 71 colorectal cancer patients with 3 M KCl extracts of cancer tissues and measured concentration of CEA in the plasma and the extracts of cancer tissue simultaneously. Although CEA in colorectal cancer extracts was individually varied from low to high in concentration, the levels of CEA had no relation to LMI reactivity. In addition, the LMI reactivity of colorectal cancer patients did not relate to the plasma CEA level of the corresponding patient. The results suggest that some antigens which induce leucocyte migration inhibition factor (LMIF) to lymphocytes from colorectal cancer patients might be different substances from CEA.

Stimulation humaner und boviner Lymphozyten durch verschiedene Streptokokken-Species

Concerning the question of species specifity of lymphokines in the presented examination the effect of the leukocyte migration inhibition factor (LIF) secreted by human and bovine lymphocytes was tested using PMN leukocytes of man and different animal species as indicator cells. The method was the so called indirect leukocyte migration inhibition test according to Clausen. As antigens for stimulating the lymphocytes cell wall preparations of group A- and B-Streptococci and as a mitogen Concanavalin A was used. From the results the following facts can be stated: — Human LIF has no inhibiting effect on the migration of leukocytes from cattle, sheep and pigs — independent of the way of stimulation by specific antigens or by the mitogen. — A corresponding factor to human LIF can be demonstrated from bovine lymphocytes which, however, inhibits only the migration of bovine indicator cells. This phenomenon is also independent of the kind of stimulation. — Migration inhibiting substances from bovine lymphocytes are only produced after stimulation with group A- and B-streptococci cell walls if the donor animals have been sensitized with these antigens. — The interaction between LIF and indicator leukocytes obviously is dependent on host specifity and not on a peculiar relation of macro- and microorganisms.

Immunoassessment of patients with gastric cancer. Using leukocyte migration inhibition (LMI) test.

Leukocytes from patients with gastric, colorectal or benign gastrointestinal disease, as well as those from normal control subjects, were tested by the leukocyte migration inhibition test (LMIT), using five different allogeneic 3 M KCl extracts of gastric cancer extracts. In LMIT with a single tumor extract, a pathological MI was found in 48% of 79 gastric cancer patients, a significantly higher figure than in the three other groups of patients (4-21%). In the panel mode of LMIT, that is, testing each blood sample with five different tumor extracts, 62/79 (79%) of patients with gastric cancer showed a positive reaction. Positive reactivity was significantly higher in patients with gastric cancer than in patients in the other groups and the frequency was higher in stage IV cases than in stage I-III cases, although the differences were not significant. In addition, the frequency of the positive reaction was unrelated to the degree of differentiation in adenocarcinoma. Positive reactivity increased in inverse proportion to the degree of nuclear grade (NG) (84% in NG I, 80% in NG II and 64% in NG III), but the correlation was not statistically significant.

Advances in the diagnosis of tuberculosis.

Several advances in the laboratory diagnosis of tuberculosis have been described recently. Refinements in collection of specimens, methods of staining and culture techniques have been described. Several newer immunological tests such as ELISA, SAFA, and leucocyte migration inhibition test are useful in the diagnosis of sputum negative pulmonary tuberculosis. Indirect immunoflouresence test and leucocyte migration inhibition test are useful in determining the duration of chemotherapy. Detection of mycobacterial cellular products such as tuberculostearic acid in sputum are helpful in diagnosing sputum negative cases. This article describes briefly the principle and usefulness of these tests in diagnosis of tuberculosis.

Acquired cellular responsiveness in cattle cleared of [formula omitted] 28 months earlier

The leukocyte migration inhibition test (LMIT) was used to assess cell-mediated immune responses of cattle against Anaplasma marginale . Leukocytes from 7 of 11 cows that had been cleared of the carrier state by antibiotic therapy 28 months earlier were inhibited from their normal migration by A. marginale antigens ( P <0.001 ). After reexposure to virulent organisms, all animals were positive to the LMIT. There was not a significant relationship (r=0.49) between LMIT values before and after reexposure. Results suggest that animals free of anaplasmosis for more than 2 years are capable of cell-mediated immune responses, despite decreased in vitro responsiveness at the time of reexposure. Acquired cellular responses did not prevent reinfection as all animals became A. marginale carriers.

Fetal gastric and colonic implants in syngeneic and allogeneic mice developing typical inflammatory changes.

Implants of fetal stomach and colon under the kidney capsule of syngeneic, and H-2 compatible and H-2 incompatible allogeneic mice were examined histologically at different time intervals after the procedure. According to the time of implantation typical inflammatory changes were seen in syngeneic stomach and colon implants, which resembled changes seen in chronic atrophic gastritis and chronic ulcerative colitis. Immunofluorescence studies showed that the host developed antibodies against fetal antigens, while there was no evidence for cellular immune response to fetal syngeneic antigens with the direct leukocyte migration inhibition test. Possible explanations for these results are discussed.

Leucocyte migration inhibition test in coeliac disease - a reappraisal.

Results of the direct leucocyte migration inhibition (LMI) test using gluten fraction III as antigen were unaffected by incorporation of puromycin into the culture medium at concentrations shown to prevent lymphokine mediated inhibition. Results of the LMI test performed with purified polymorphs were similar to and correlated with results of the standard LMI test using mixed leucocytes in both coeliacs and controls. The addition of purified T lymphocytes did not increase migration inhibition. Normal leucocytes incubated with serum from coeliac patients and washed showed marked migration inhibition when incubated with gluten fraction III. This sensitisation of normal leucocytes was prevented by preincubation with aggregated human IgG. These results suggest that leucocyte migration inhibition by gluten in coeliac disease is not due to lymphokine production by sensitised lymphocytes but is caused by cytophilic antibody.

Cell-mediated immunity to myoglobin in polymyositis.

Using the leucocyte migration inhibition (LIF) test we looked for evidence of cell-mediated hypersensitivity against myoglobin in 8 patients with polymyositis (PM) or dermatomyositis (DM). The migration index for PM-DM patients was 47.5 +/- 17%, while in the controls the index was 86 +/- 12% (p less than 0.001). The presence of serum antibodies against myoglobin was also investigated by passive haemagglutination (PH) and counterimmunoelectrophoresis (CIE). By PH the reciprocal titre of antimyoglobin antibodies was 150 +/- 28 in PM-DM compared with 16.2 +/- 15.9 and 8.7 +/- 7.7 in control patients with diseases (p less than 0.01) and negative in normal persons. CIE showed antimyoglobin antibodies in 3 PM-DM patients and in none of controls. Cell-mediated immunity against myoglobin may be implicated in the pathogenesis of PM. The pathological significance of antimyoglobin antibodies remains to be determined.

Diagnosis of penicillin allergy

A study was made to establish the value of the leucocyte aggregation test (LAT) in drug allergy using penicillin antigen. The antigen-induced human peripheral blood leucocyte aggregation was measured quantitatively. The results obtained have been compared with the leucocyte migration inhibition test (LMIT) in patients with or without delayed penicillin allergy. Among forty-four penicillin-allergic subjects and thirty-six control subjects, LAT was found positive in respectively 70.5 and 30.5% (P less than 0.001) whereas LMIT was found positive in respectively 56.8 and 50% of the patients. These results were confirmed by multiple correspondence analysis (MCA), using a computer. Furthermore, this method, enables a more comprehensive and reliable interpretation of the tests, by the help of various quantitative and qualitative criteria. It is concluded that LAT shows more discrimination than the LMIT in distinguishing a penicillin-allergic population from a non-allergic one. In addition, LAT offers great technical advantages over the LMIT for the diagnosis of drug allergy.