E. Clinton Lawrence

Immunoglobulin Secreting Cells in Normal Human Bronchial Lavage Fluids

Immunoglobulin secreting cells were quantitated in the bronchial lavage fluids of 12 normal volunteers and compared with immunoglobulin secreting cells in peripheral blood, by a reverse hemolytic plaque assay. The mean number of cells secreting immunoglobulin (Ig)G in bronchial lavage fluids was 489 per million lymphocytes vs. a mean of 175 IgG secreting cells per million lymphocytes in peripheral blood (P < 0.02). The mean number of IgA secreting cells in bronchial lavage fluids was 633 per million lymphocytes as compared to 100 per million lymphocytes in peripheral blood (P < 0.005). Thus, compared to peripheral blood, cells from the lavage fluids were relatively enriched for both IgG and IgA secreting cells. However, IgA secreting cells were the major class of immunoglobulin secreting cells in bronchial lavage fluids, whereas IgG secreting cells predominated in peripheral blood. The prominence of IgA secreting cells in bronchial lavage fluids was further demonstrated by a mean ratio of IgA/IgG secreting cells in bronchial lavage fluids of 1.26 compared to a ratio in peripheral blood of 0.57 (P < 0.02). Cells secreting IgM were identified in only four of seven bronchial lavage fluid samples studied but in all peripheral blood samples. IgE secreting cells were not present in normal peripheral blood but could be demonstrated in 5 of 11 lavage fluid specimens. Thus, cells actively secreting immunoglobulins can be identified in the lower bronchial-alveolar tree of normal human subjects. Cells secreting IgG, IgA, or IgM may function in local lung defenses against infection; cells secreting IgE may contribute to hypersensitivity reactions in the lung.

Serial changes in markers of disease activity with corticosteroid treatment in sarcoidosis

Serial changes in various markers of disease activity with corticosteroid therapy were assessed in 12 patients with active sarcoidosis. After six weeks of treatment with 40 mg daily of prednisone, all but one patient demonstrated symptomatic and radiographic improvement. For the entire patient group, there were corresponding improvements in forced vital capacity, from 59.2 +/- 5.5 to 70.5 +/- 5.3 percent of the predicted value (p less than 0.001, Student paired t test), serum angiotensin-converting enzyme levels, from 66.0 +/- 12.1 to 28.2 +/- 4.0 U/ml (p = 0.003), 67gallium lung scanning scores, from 3.6 +/- 0.2 to 0.8 +/- 0.3 (p less than 0.001), serum gamma globulin levels, from 2.40 +/- 0.2 to 1.5 +/- 0.1 g/dl (p less than 0.001), and erythrocyte sedimentation rate, from 26.8 +/- 2.7 to 14.8 +/- 3.0 mm per hour (p less than 0.001). Changes in percent of bronchoalveolar lavage fluid lymphocytes were less impressive (from 28.7 +/- 4.9 to 21.2 +/- 5.1, p = 0.034), but the geometric mean number of bronchoalveolar lavage fluid-IgG-secreting cells decreased from 23,861 to 3,830 (p = 0.013). Serial evaluations in five patients treated with decreasing doses of alternate-day prednisone for an additional 10 1/2 months indicated that changes in 67gallium lung scanning scores corresponded most closely to the clinical course in five of five patients. Determination of serum angiotensin-converting enzyme levels also closely paralleled the clinical course in four of five patients, whereas the other parameters measured were more variable markers of clinical response. However, abnormalities of bronchoalveolar lavage fluid-IgG-secreting cells often persisted in the absence of clinically evident disease, and the percentages of bronchoalveolar lavage fluid lymphocytes were frequently normal in patients who responded subsequently to corticosteroids. Larger prospective studies are warranted to more extensively evaluate various measurements of disease activity, especially bronchoalveolar lavage fluid analysis, in sarcoidosis.

Cyclosporine therapy of central nervous system sarcoidosis

With increasing numbers of patients covered under diagnostic-related group or prospective payment systems, there is mounting pressure to reduce the costs associated with caring for patients while maintaining the quality of care. New, broad-spectrum, parenteral antimicrobial agents are a prime target for cost control. The availability of an orally administered drug that is effective in a variety of serious infections would have a significant impact on cost reduction in hospitals. A significant proportion of the cost of parenteral drug therapy is associated with labor, supplies, and equipment. An orally administered drug would obviate these costs. Additionally, iatrogenic problems associated with parenteral therapy (e.g., phlebitis) would be avoided, the quantities of expensive parenteral drugs purchased might be reduced, and the duration of hospitalization for selected diseases such as osteomyelitis would probably decrease. Finally, the ease of oral administration may allow longer courses of therapy for resistant infections than have heretofore been possible. Oral ciprofloxacin and similar compounds present both opportunities and challenges for pharmacists. The fiscal impact may be beneficial for the hospital in terms of cost reductions for drug purchases and in expediting discharge for prolonged courses of outpatient therapy. The magnitude of cost reductions will vary substantially depending upon the types of patients and infections managed in a given institution. The challenge for pharmacists is to assure that these valuable compounds, as well as all antimicrobial agents, are appropriately utilized to avoid the offsetting costs of adverse effects, superinfection, unnecessary use, and the development of resistance.

Permanent intrinsic B cell immunodeficiency caused by phenytoin hypersensitivity

We report a patient who, 3 weeks after initiation of therapy, experienced a hypersensitivity reaction to phenytoin manifested as rash, lymphadenopathy, elevated serum transaminase levels, and subsequent panhypogammaglobulinemia with IgG, 180 mg/dl (control range 639 to 1349); IgA, 15 mg/dl (control range 70 to 312); and IgM, 0 mg/dl (control range 56 to 352). Repeated in vitro lymphocyte analysis documented normal T cell-mediated immunity including T cell surface markers (E rosettes), lymphocyte proliferation after mitogen stimulation, and T cell phenotypes (T4 or helper and T8 or suppressor cells). However, the patient had a paucity of circulating B-lymphocytes as assessed by the number of lymphocytes with surface membrane immunoglobulin (patient value of 0 compared to the control range 16 to 435 cells per microliter of blood) and by the number of lymphocytes bearing the B1 antigen (patient value of 13 compared to the control range 48 to 358 cells per microliter of blood). Hemolytic plaque assay revealed decreased immunoglobulin production (number of immunoglobulin-secreting cells per million circulating mononuclear cells) as compared to control subjects (patient unstimulated mean of 100 as compared to a control mean of 1753) and minimal enhancement on stimulation with pokeweed mitogen (patient stimulated mean of 250 as compared to control mean of 8946). Coculture experiments with the reverse hemolytic plaque assay revealed no evidence of suppression. No reversal of this patients immunodeficiency has occurred 3 years after phenytoin withdrawal.

Imbalances in subsets of T lymphocytes in an inbred pedigree with Omenn's syndrome

Abstract The subsets of T lymphocytes in two infants affected with Omenns syndrome and in 109 healthy members of the highly inbred pedigree were studied. Eighteen homozygotes in this pedigree had previously died from infection at less than 6 months of age. Both infants displayed normal numbers of peripheral blood T (E-rosette) lynphocytes, poor mitogen reactivity of lymphocytes, normal mixed lymphocyte culture reactivity, a paucity of circulating B cells, variable hypogammaglobulinemia, and elevated serum IgE concentrations. At 4 months of age, one infant (boy) had 95% T3+ (total T) peripheral blood lymphocytes, 41% T4+ (helper T) lymphocytes, and 64% T8+ (suppressor T) lymphocytes; at 4 months of age the other infant (girl) had 43% T3+, 43% T4+, 19% T8+ lymphocytes, and 18% T6+ (stage II thymocyte) lymphocytes. Age-matched controls had values of 49% for T3+, 37% for T4+, 13% for T8+, and

Cigarette Smoking and Bronchoalveolar T Cell Populations in Sarcoidosis

Pulmonary physicians must often deal with patients, including patients with sarcoidosis, who smoke cigarettes. Since changes in local pulmonary immune function have been associated with both sarcoidosis’4 and cigarette smoking;-’o it is important to distinguish which of these immunological changes in the lungs are due to the disease, which are due to cigarette smoking, and which, perhaps, are due to both. Abnormally large numbers of helper thymus-derived (T) lymphocytes are found in fluids recovered by bronchoalveolar lavage (BAL) from patients with sarcoidosis.“ By contrast, normal numbers of lymphocytes are found in BAL fluids from normal cigarette smokers,6 and lower than normal percentages of the cells in these fluids are lympho~ytes .~*~~’~ The effects of smoking on T lymphocyte subpopulations in normal cigarette smokers and on those in patients with sarcoidosis, however, have not been thoroughly characterized. The purposes of this study, therefore, were 1) to determine the effects of smoking on T lymphocyte subpopulations in BAL fluids from healthy normal volunteers (“normals”) and on those in BAL fluids from patients with sarcoidosis ((‘sarcoids”), and 2) to determine whether comparisons of the T lymphocyte subpopulations of normals and those of sarcoids revealed any effects of cigarette smoking.

Asbestos body phagocytosis by human free alveolar macrophages.

Phagocytosis of asbestos bodies by human free alveolar macrophages (FAMs) was documented employing light microscopy. This process was more carefully studied utilizing scanning electron microscopy (SEM) which demonstrated morphological and surface membrane changes in FAMs following phagocytosis of asbestos bodies. FAM viability was also evaluated following 24--72-h incubation of cells with asbestos bodies at a final concentration of 250 micrograms/ml. Slight, but significant, cytotoxicity was observed following the initial 24-h culture period (P = 0.032, paired, 2-tailed t-test). No further cytotoxicity was observed, however, when cells were further incubated for 48-h and 72-h intervals (P greater than 0.05 in all instances). These studies demonstrate asbestos bodies are readily phagocytized by cultured FAMs, and are only slightly cytotoxic to these human lung cells.

In vitro cytotoxicity of chrysotile asbestos to human pulmonary alveolar macrophages is decreased by organosilane coating and surfactant

Human pulmonary alveolar macrophages were used to quantitate the cytotoxic effect of surface-altered chrysotile asbestos. Little difference was observed in mortality between chrysotile asbestos that was surface-treated to a 42% extent by a hydrophobic organosilane or untreated chrysotile. Little or no effect on mortality was observed when human pulmonary alveolar macrophages were cultured with untreated chrysotile or acid-leached asbestos in the presence of 10 mM dipalmitoyl lecithin. However, when human pulmonary alveolar macrophages were cultured with a hydrophobically-treated (to a 42% or 95% extent) chrysotile asbestos in the presence of 10 mM dipalmitoyl lecithin, a statistically significant decrease in mortality was observed compared to untreated chrysotile. No mutagenic activity was observed when V79 cells were cultured with acid-leached, or 42% hydrophobically-treated chrysotile asbestos, even when human pulmonary alveolar macrophages were included as an activation source. The 95% hydrophobically-treated and acid-leached chrysotile also exhibited decreased binding of benzo[a]pyrene compared to untreated chrysotile asbestos.

902 MONOCYTE SUPPRESSION OF IMMUNOGLOBULIN SECRETION IN A 9-YEAR-OLD BOY WITH SCID

We report the evaluation of in vitro immunoregulation in a 9-year-old untreated boy with SCID. Ficoll-Hypaque-isolated peripheral blood mononuclear cells (MNL) from the patient failed to respond to pokeweed mitogen (PWM) with the normal increment in immunoglobulin-secreting cells (Ig-SC), as measured by a reverse hemolytic plaque assay. Removal of phagocytic cells or the addition of unrelated irradiated helper cells resulted in enhanced, but still suboptimal response to PWM, suggesting some intrinsic defect in B-cell function. Co-culture of patient with normal MNL resulted in marked (88%) suppression of PWM-induced Ig-SC. Suppressor activity was unaffected by prior irradiation of patient MNL, but was substantially reversed (32% net enhancement) by removal of his phagocytic cells; whereas the combination of the two procedures further reversed suppression (52% net enhancement). Because the patient was lymphopenic, his MNL were relatively enriched for monocytes (range=40-80%). To determine whether the suppressor cell activity was due to functional or numerical excess of patient monocytes, co-cultures were performed using varying ratios of patient to normal MNL. Suppression was most marked in cultures containing >40% monocytes, suggesting that a numerical excess of patient monocytes could account for the observed suppression. These data suggest both an intrinsic defect in B-cell function, and a relative numerical excess of monocytes which could further inhibit Ig-secretion by B-cells.