An immunological approach to the diagnosis of food sensitivity.
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The numbers of eosinophils recruited locally to abraded human skin were measured in eight atopic and eight non-atopic volunteers, at time intervals over 24 hr, following the application of an ECF-A tetrapeptide (Val-Gly-Ser-Glu) or histamine. In all subjects the higher doses of the peptide (10(-4) and 10(-6) mol/l) or histamine (10(-3) and 10(-5) mol/l) produced significantly greater counts than the Tyrode's diluent alone. The counts produced with the lowest dose of peptide (10(-8) mol/l) or histamine (10(-7) mol/l) were not significantly different from the control. The peptide or histamine evoked a greater local eosinophilia in the atopics than the non-atopics. This effect was probably independent of the peripheral blood eosinophil counts since at the time of study the numbers of circulating eosinophils between the two groups were not significantly different. In the atopics, histamine in doses of 10(-3) and 10(-5) mol/l were required to give the same eosinophil response as that obtained with 10(-4) and 10(-6) mol/l of the peptide, respectively. It is suggested that the relative paucity of eosinophils recruited by locally applied ECF-A peptide or histamine, when compared to antigen-induced eosinophilia, is due either to an inability to mimic the events associated with the release of these mediators from mast-cells or the involvement of other pharmacological agents.
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Healthy children of atopically asthmatic parents received in vivo and in vitro granulocyte migration tests. Eleven children whose parents were both asthmatic showed statistically significant inhibition of granulocyte migration compared with controls.
Neutrophil migration from skin window abrasions was studied in 10 patients with no history of periodontitis, 10 with localised juvenile or post-juvenile periodontitis, and 10 with chronic adult periodontitis. Filters contained either saline or chlorhexidine (0.002% or 0.02%). The leading front was measured in filters placed for 30 min after cell migration had been established for 2 h. Subjects in the juvenile/post juvenile group showed a reduced range of migration distances, but were still within the normal range when compared with the other 2 groups. Chlorhexidine at 0.002% tended to increase leading front distances, and 0.02% to decrease them. We conclude that: 1) migrating neutrophils in vivo may move less far in patients with a history of juvenile periodontitis; 2) chlorhexidine may inhibit cell migration, possibly decreasing the host response in vivo if applied at current therapeutic concentrations.
Cellular immunity has been studied by means of lymphocyte response to PHA, delayed hypersensitivity and skin window responses in 23 patients with myeloma (14 IgG, 9 IgA) and 14 patients with macroglobulinaemia. In the myeloma patients, 14% had abnormal PHA response and 29% were anergic. In those with macroglobulinaemia, 29% showed abnormal PHA response and 57% were anergic. In myeloma, the abnormal PHA response was due to a serum inhibitor. Abnormal skin window responses were present in 75% of the patients with myeloma, but only 22% of those with macroglobulinaemia. All the myeloma patients with anergy had abnormal skin windows but this correlation did not exist in macroglobulinaemia. No correlation was found between the paraprotein concentration and anergy, PHA response or skin window. The results support the conclusion that myeloma is predominantly associated with an abnormal skin window (inflammatory) response and macroglobulinaemia with intrinsic abnormalities of cellular immunity. When anergy and abnormal PHA response are present in myeloma, it appears to be attributable to an effect of the paraprotein and not an intrinsic abnormality of lymphocytes.
Ten patients with far-advanced Laennec's cirrhosis were studied for in vitro polymorphonuclear (PMN) chemotaxis and in vivo PMN delivery into modified skin windows. Random motility was similar for patient and control cells, but the patients' cells' mean chemotaxis to endotoxin-activated homologous serum was only 38% of control. The poor response was due to a serum inhibitor which reduced chemotactic activity of control serum by 75.6% but had no effect on chemotaxis to N-formyl-methionyl-leucyl-phenylalanine. The level of inhibitor activity in each patient did not correlate with the severity of liver dysfunction. In spite of the universal presence of the chemotactic factor inhibitor in all patients' sera, their in vivo PMN delivery into skin chambers was similar to controls. Thus, patients with alcoholic cirrhosis appear able to deliver PMNs normally in response to local tissue injury.
This study investigated the effect of a chemoattractant, N-formyl-methionyl-leucyl-phenylalanine (FMLP), upon skin window migration of neutrophils into filters in 5 patients with a history of localized juvenile periodontitis (LJP) and 8 controls. On 2 occasions, each subject had 2 superficial skin abrasions made on the inner aspect of the forearm. Initial periodontal treatment was carried out on the disease group between the visits. On one skin window filters were placed that were soaked in physiological saline, and on the other filters soaked with FMLP. Leading fronts and cell densities were measured in each filter. At visit 1, LJP subjects had significantly lower leading fronts and cell densities. At visit 2, the differences were insignificant. The leading fronts for the LJP group were significantly improved on the second visit. No difference was observed between saline and FMLP. The findings of this study indicate that neutrophil migration is reduced in LJP patients where treatment is not involved, and that FMLP has no effect on neutrophil migration from the skin windows under the conditions of this study.
Examination of the blood of 40 patients with leukaemic reticuloendotheliosis (LRE) revealed a drastic reduction in monocyte numbers (mean 74/microliter) when compared to a normal group of 33 (mean 442/microliter). Repeated blood studies in 8 LRE patients were made over periods of 2 months to 5 years and monocytopenia was observed to be a persistent phenomenon. Similarly, tissues from 29 patients showed an overall decrease in numbers of monocytes/ histiocytes when compared with those of 73 control subjects. Skin window examinations in all 11 patients so studied showed marked reduction or absence of monocyte response to inflammation. 2 exceptional cases to this trend are noted. In one, circulating monocyte levels remained normal over a 14-month period while examinations of blood and marrow in this case showed few hairy cells. In another case, severe and persistent monocyte deficiency was noted during a 2-year period of unsuccessful treatment. When he eventually responded to low dose chlorambucil and his Hb rose from 40-140 g/l and WBC became normal, the number of circulating monocytes also became normal.
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We investigated 582 cases of sarcoidosis by the skin-window (Rebuck) method. The number of macrophages appearing in skin windows was significantly increased in 432 patients with sarcoidosis in the active phase. Further, increased numbers of macrophages in the windows were found in patients with a positive Kveim-Siltzbach test, high immunoglobulin levels, increased B lymphocytes, and decreased T lymphocytes. Passive transfer of delayed-type hypersensitivity was weaker and shorter in sarcoidosis patients with increased presence of macrophages than in healthy controls. A close correlation between high macrophage reactivity and the other sepcific immunological factors in sarcoidosis is suggested.
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