Properties and activities of transfer factor

Although there is agreement that transfer factor endows skin test-negative subjects with the ability to develop the delayed allergic responses of the transfer factor donors, there is little direct information on the mechanism of this phenomenon or on the nature of the active components (s). This report reviews some of the known effects of transfer factor or immune responses and inflammation. It is concluded that transfer factor has multiple sites of action, including effects on the thymus, on lymphocyte-monocyte and/or lymphocyte-lymphocyte interactions, as well as direct effects on cells in inflammatory sites. It is also suggested that the "specificity" of transfer factor is determined by the immunologic status of the recipient rather than by informational molecules in the dialysates. Finally, it is proposed that many effects of transfer factor may be due to changes in intracellular cyclic nucleotide content, especially accumulation of cGMP, in immunologically reactive cells.     

Renal arteriography following systemic reaction to contrast material

A 15-year-old atopic female with severe hypertension (B.P. 220/160) was evaluated for evidence of renal arterial disease with equivocal results, and the performance of renal arteriography was considered essential. However, she had a history of two well-documented anaphylaxis-like reactions to contrast materials, considered to be an absolute contraindication to the performance of such a study. When the etiology of this sensitivity was investigated, an IgE-dependent mechanism was not evident. However, the patient's leukocytes released significantly greater quantities of histamine upon incubation with contrast materials than did 7 of 9 normal volunteers. The 2 hyperresponsive normal subjects were not atopic and had no prior exposure to contrast materials. Pretreatment with 80 mg prednisone and 200 mg diphenhydramine daily for 3 days prior to arteriography and challenge with increasing quantities of intravenous iothalamate meglumine (Conray) at the time of the study resulted in successful performance of arteriography and identification of fibromuscular dysplasia of the right renal artery. Placement of an aortorenal bypass graft corrected her hypertension. Thus, arteriography can be safely performed in selected patients with severe previous reactions to contrast materials when the alternatives provide no lesser risk by pretreatment with a steroid-antihistamine regimen and gradual administration of increasing doses of the contrast agent.     

Immunologic reactivity in the hypereosinophilic syndrome.

Because previous studies have suggested an important link between eosinophilia and immunologic reactivity, we investigated various components of the immune system in a large number of patients with the idiopathic hypereosinophilic syndrome (HES) to elucidate a possible role for immunologic phenomena in the etiology and pathogenesis of this disease. Immunoglobulin G, A, or M levels were only rarely abnormal. However, in 8 of 21 (38%) patients with HES, IgE levels were markedly elevated suggesting an association of an IgE-mediated mechanism with eosinophilia in this subgroup. Severe dermatographism was present in three fourths of patients, and 2 patients with intermittently elevated histamine levels manifested an unusual form of immediate-pressure urticaria. Serum complement determinations showed elevated C4 and C3 levels in 27% and 77% of patients, respectively. Antigen-antibody complexlike material measured by C1q binding was elevated in the serum of 7 of 22 (32%) patients; this finding may relate to the known ability of eosinophils to avidly phagocytose antigen-antibody complexes. When compared with normals, lymphocytes from patients with HES showed a variety of abnormalities of lymphocyte surface receptors and lymphocyte function. Thus, patients with HES demonstrate a variety of immunologic abnormalities which may be related primarily or secondarily to the pathogenesis of this syndrome.     

Study of possible mechanisms of basophil accumulation in experimental cutaneous candidiasis in guinea pigs

It is known that certain lymphokine preparations, bacterial growth products, and factors released through complement activation have in vitro chemotactic activity for basophils. We have developed a model for acute cutaneous candidiasis in guinea pigs in which the lesions are characterized by infecting organisms in the keratin layer, early accumulation of polymorphonuclear leukocytes in the upper epidermis, and subsequent accumulation of basophils along the dermal basement membrane. The present study was undertaken to determine if any of the known chemotactic factors were operating in vivo to attract basophils. Both nonimmune guinea pigs and animals with established delayed hypersensitivity to candida had basophils in the infected skin. While immune animals showed more basophils than did nonimmune animals, the difference was not significant. Intradermal injections of a sonicate of candida or a candida growth filtrate did not cause significant accumulation of basophils. Decomplementation of the animals with cobra venom factor (CVF) did not significantly reduce the basophil numbers. Moreover, basophil accumulation occurred in animals with only minimal serum antibody to candida. These studies indicate that the basophil accumulation is due to a mechanism that is not dependent on cellular immunity, direct chemotactic activity in the candida extract, antibodies, or complement. Therefore, there may exist a previously unrecognized, nonimmunologic mechanism of chemotaxis for basophils which could possibly operate in other types of lesions and could even be involved with attraction of other types of cells.     

An approach to the use of immunosuppressive drugs in nonmalignant diseases

Immunosuppressive drugs are being given to patients with inflammatory diseases of unknown etiology. The drugs in common use have different biochemical properties that may be responsible for their different immunologic and anti-inflammatory properties. These are reflected in clear-cut distinctions in clinical efficacy. A limited number of controlled trials have suggested that certain immunosuppressive drugs are effective in a few human inflammatory diseases. There have been more negative than positive trials. Furthermore, the drugs increase the risk of infection and may predispose to malignancy. Long-term results are not available; until they are, the drugs would be most profitably administered in the context of controlled long-term trials so that rational approaches to therapy may be possible in the near future. Important medicophilosophical questions are raised by the potential benefit and the potential harm of immunosuppressive drugs in the treatment of nonmalignant diseases. The widespread use of immunosuppressive drugs in the therapy of human diseases of unknown etiology is not warranted at the present time.     

Assessment of tissue fluid histamine levels in patients with urticaria

Tissue fluid histamine in lesions and normal skin sites of patients with various forms of physically induced urticaria and chronic idiopathic urticaria was assessed utilizing suction-induced blisters. Histamine levels were elevated over challenged sites in cold urticaria, solar urticaria, and delayed pressure urticaria. Histamine levels were elevated in both challenged and “control” sites in patients with immediate-pressure urticaria and local heat urticaria in whom the apparatus provoked lesions. In 5 of 13 patients with chronic idiopathic urticaria, the blister fluid histamine levels were elevated in lesions but not in normal-appearing skin, while in 7 patients the blister fluid histamine levels were elevated in lesions as well as in normal-appearing skin. Although the pathogenesis of chronic idiopathic urticaria is unknown, a clear abnormality related to skin histamine has been identified.     

In vivo studies of mediator release in cold urticaria and cholinergic urticaria

Six patients with cold urticaria were found to possess elevated plasma histamine levels after cold challenge by placing one hand in ice water for 4 minutes. A single patient became hypotensive during the procedure and had a level of 260 ng/ml. histamine in the venous effluent from his hand. No elevation of plasma serotonin or bradykinin was observed. Two patients with cholinergic urticaria possessed elevated plasma histamine levels during and after vigorous exercise for 10 minutes; these patients also gave a positive test for vibration-induced angioedema. A single patient with cholinergic urticaria possessed elevated baseline serotonin levels and elevated levels during and after exercise but no elevation of plasma histamine or bradykinin. The results suggest that histamine is the major mediator of urticaria and hypotension in cold urticaria. Histamine also appears to be released coincident with the development of urticaria in some patients with cholinergic urticaria, while elevated serotonin levels in a single atypical patient suggest that a subpopulation of patients with cholinergic urticaria possess a different pathogenesis.     

Automated histamine analysis for in vitro allergy testing. II. Correlation of skin test results with in vitro whole blood histamine release in 82 patients.

The recently developed sensitive, automated histamine assay system was applied for in vitro allergy testing. The simplified method for histamine release from whole heparinized blood was used. Aliquots of blood and allergen were incubated for one hour at 37 degrees C, and each supernatant was then analyzed for histamine release. Nine common pollen and environmental allergens were used at three 10-fold dilutions for in vitro testing with the use of 20 ml of blood. Intradermal skin tests were correlated with the whole blood histamine release in 82 patients who had received no immunotherapy. A scoring system for the histamine results was developed to take into consideration the results with multiple allergen concentrations. When the skin test was strongly positive (greater than or equal to 3 + at 100 protein nitrogen units [PNU]/ml), the whole blood histamine release was positive in 89% of the tests. In contrast, when the skin test was negative ( less than 1 + at 100 PNU/ml), the histamine release was also negative in 99.8% of the cases. When the skin test was 1 +, the histamine release from whole blood was positive in 6% of the tests; and when the skin test was 2+, the whole blood results were positive in 32%. The accuracy, precision, and sensitivity of the automated histamine assay allow its application for the clinical study of allergic patients.     

Human lung tissue and anaphylaxis. I. The role of cyclic GMP as a modulator of the immunologically induced secretory process.

A close relationship between increased concentrations of cyclic GMP in human lung tissue and the capacity for acetylcholine to enhance the immunologic secretion of histamine and SRS-A has been found. Acetylcholine (10(-7) to 10(-11) M) produced parallel increases in both cyclic GMP and the immunologic release of mediators; the muscarinic blocking agent atropine prevented both responses. The increase in cyclic GMP in human lung after acetylcholine stimulation was apparent within 30 sec, peaked by 120 sec, and abruptly returned to control levels thereafter. The ability of acetylcholine to enhance the antigen-stimulated secretion of mediators followed the same time-course. PGF2alpha (3.3 X 10(-4) M to 3.3 X 10(-7) M) increased the cyclic GMP content of human lung tissue in a dose-related fashion. Pretreatment of IgE-sensitized lung tissue with acetylsalicylic acid (10 microgram/ml) had no effect on baseline cyclic nucleotide levels, the capacity for antigen to induce mediator release, or the increase in cyclic GMP and facilitation of the immunologic release of mediators produced by acetylcholine.     

The effects of histamine and prostaglandin D2 on rat mast-cell cyclic AMP and mediator release

The possibility that histamine may play a functional role in modulating mast-cell secretion, as has been suggested for basophil degranulation, has both physiologic and pharmacologic implications. Therefore the capacity of histamine to influence rat peritoneal mast-cell (RPMC) cyclic AMP levels and reversed anaphylatic degranulation as reflected in the release of 3H-serotonin (5-HT) was examined. To ascertain that RPMC were functionally responsive to exogenous hormonal stimulation, assessment of prostaglandin (PG) D2 effects on cyclic AMP and 5-HT release were determined in parallel. Although PGD2 (100 microM) increased cyclic AMP and inhibited 5-HT release in the presence of 50 microM aminophylline, histamine (up to 1000 microM) was ineffective was ineffective in both. However, 1000 microM histamine in the presence of 500 microM aminophylline was capable of transiently increasing RPMC cyclic AMP (for 15 to 30 sec) and under these conditions of suppressing 5-HT release. The receptor subtype involved in the suppressive actions of histamine appeared to be of the H-1 type as reflected in the capacity of specific H-1 agonists to reproduce the inhibition of 5-HT release, whereas neither H-2 agonists nor H-2 antagonists had any influence. Thus, under conditions in which phosphodiesterase enzymatic action is impaired, histamine in extremely high concentrations is able to modulate mast-cell secretion. However, it seems very unlikely that this action of histamine has any physiologic significance.     

Measurement of urinary histamine: Development of methodology and normal values

A small portion of the histamine that circulates through the kidney is excreted intact. Thus, the measurement of histamine in urine may be employed to monitor fluctuations in plasma histamine and has several advantages: stability, accessibility, and the opportunity for retrospective analysis. A method for measuring urine histamine was developed based on cation-exchange chromatography, organic solvent extraction, o-phthalaldehyde condensation, and measurement of fluorescence. However, because the histamine measured by this procedure was higher than that measured by other techniques, a portion of each sample was digested with diamine oxidase and the difference between the two portions of each sample, after isolation and fluorescent assay, was taken to reflect histamine. Normal urinary histamine levels of 13 ± 8 ng/ml, 14 ± 9 μg/24 hr, or 14 ± 72 ng/mg creatinine/ml were found. Male and female subjects excreted equivalent concentrations; spot, short-timed, or 24-hr collections provided equivalent results; and histamine in frozen urine was stable ?6 mo. Two patients with systemic mastocytosis and two with idiopathic anaphylaxis had elevated urine histamine levels. Monitoring urine histamine may be useful in assessment of conditions in which histamine plays a role.     

Tropical eosinophilia : A human model of parasitic immunopathology, with observations on serum IgE levels before and after treatment

The diverse clinical syndromes characterized by asthmatic symptoms, transient pulmonary infiltrates, and eosinophilia have tended to obscure the specific association of one such entity with filarial infections. Serum IgE levels were determined before and after therapy in a group of well-characterized patients with tropical eosinophilia (TE), studied earlier in Singapore. The mean serum IgE level in 14 cases before treatment with diethylcarbamazine was 2,355 ng. per milliliter, with a trend but statistically nonsignificant decrease in levels to 600-1,000 ng. occurring 8 to 12 weeks after therapy. Leukocyte and eosinophil counts showed a rapid reduction after treatment, and although mean complement-fixing (cf) titers to Dirofilarial antigen tended to decrease, they were not significantly reduced until 5 to 6 weeks. The historical development of evidence supporting the filarial etiology of TE was reviewed. Many basic questions engendered by the clinical syndrome of tropical eosinophilia make it an excellent model for study of the immunopathology of parasitic infections.     

Abnormal beta adrenergic responsiveness in allergic subjects: analysis of isoproterenol-induced cardiovascular and plasma cyclic adenosine monophosphate responses

Beta adrenergic responses were compared between normal control subjects and patients with allergic asthma or allergic rhinitis and a group of asymptomatic individuals with positive immediate hypersensitivity skin tests. Two responses to infused isoproterenol were monitored: increases in pulse pressure and plasma cyclic adenosine monophosphate (cyclic AMP). Although the normal control subjects responded with increases in pulse pressure of greater magnitude than the other groups in response to infused isoproterenol, the differences in responsiveness were more apparent when the concentration of isoproterenol required to increase the pulse pressure ≧22 mm Hg was compared. The 25 control subjects required 8.04 ± 0.48 ng/kg/min isoproterenol, whereas the 17 asthmatic patients required 14.25 ± 1.21 (p < 0.0001), the eight subjects with allergic rhinitis required 12.75 ± 1.58 (p < 0.0005), and the seven asymptomatic subjects with positive skin tests needed 11.1 ± 1.26 (p < 0.01). Comparison of baseline plasma cyclic AMP levels of the groups revealed no apparent differences. However, the normal controls responded to isoproterenol with larger increases in plasma cyclic AMP than the other groups at both 6 and 9 ng/kg/min. Furthermore, significantly fewer of the subjects with asthma, rhinitis, or positive skin tests demonstrated an increase in plasma cyclic AMP ≧ 50% above baseline as compared with controls. Thus, as reflected in both cardiovascular and cyclic AMP responses to infused isoproterenol, allergic asthmatic subjects are hyporesponsive as compared with normal controls. However, a comparable degree of diminished responsiveness is seen in subjects with allergic rhinitis or in asymptomatic individuals with positive skin tests. Therefore beta adrenergic hyporesponsiveness appears to be related with the atopic state rather than with asthma.     

Dopamine inhibition of histamine-mediated cutaneous responses

Several patients receiving dopamine for hypotension were skin tested for possible penicillin sensitivity. Not only were the penicillin skin tests negative but also the histamine control. On the possibility that dopamine might affect cutaneous histamine responses, we examined the effect of dopamine on histamine, antigen, morphine, and compound 48/80 skin responses. Both intradermal and intravenous dopamine selectively inhibited histamine but not antigen, morphine, or compound 48/80 skin responses, and the inhibition was in a dose-related fashion. This observation indicates that histamine should not be used to demonstrate dermal reactivity in patients receiving dopamine. The results of this study also suggest that histamine may not be the sole mast cell-derived mediator involved in the wheal-and-flare reaction characteristic of immediate-type skin tests since dopamine did not affect skin reactions caused by endogenous mast cell degranulation. Finally, the possible use of dopaminergic drugs in diseases with histamine-associated symptoms is discussed.     

Prostaglandin generation by human and guinea pig lung tissue: comparison of parenchymal and airway responses.

Anaphylaxis of human lung is accompanied by the synthesis of prostaglandins (PG), including PGF2 alpha and PGE. In an analysis of the tissue source of these prostaglandins, parenchymal preparations of both human and guinea pig (GP) lungs were compared. Peripheral, relatively airway-free preparations of human lung generate PGF2 alpha and PGE in response to histamine and 2-methylhistamine, on H1 agonist, but not to dimaprit, an H2 agonist. GP parenchymal preparations respond in a similar fashion. Stimulation of these same preparations with KCl or carbachol caused no increase in the synthesis of either PG. In human airway preparations all three agonists (histamine, KCl, and carbachol) caused the selective generation of PGE. However, stimulation of GP airway preparations with the agonists caused the production of both PGE and PGF2 alpha. These data indicate that (1) human and GP peripheral lung tissues respond to H1, but not H2, stimulation with the generation of PGF2 alpha and PGE; (2) these parenchymal responses are specific and may not be attributed to muscle contraction; and (3) stimulation of muscle contraction in human airway preparations results in the selective generation of PGE while GP airways produce both PGE and PGF2 alpha.     

Measurement of plasma histamine: description of an improved method and normal values

The single isotopic-enzymatic assay of histamine was modified to increase its sensitivity and to facilitate measurement of plasma histamine levels. The modification involved extracting ³H-1-methylhistamine (generated by the enzyme N-methyltransferase acting on histamine in the presence of S-[methyl-³H]-adenosyl-l-methionine) into chloroform and isolating the ³H-l-methylhistamine by thin-layer chromatography (TLC). The TLC was developed in acetone: ammonium hydroxide (95: I0), and the methylhistamine spot (R_f= 0.50) was identified with an o phthalaldehyde spray, scraped from the plate, and assayed in a scintillation counter. The assay in plasma demonstrated a linear relationship from 200 to 5000 pg histamine /ml. Plasma always had higher readings than buffer, and dialysis of plasma returned these values to the same level as buffer, suggesting that the baseline elevations might be attributable to histamine. However, all histamine standard curves were run in dialyzed plasma to negate any additional influences plasma might exert on the assay. The arithmetic mean (± SEM) in normal plasma histamine was 3I8.4 ± 25 pglml (n = 5I), and the geometric mean was 280 ± 35 pg/ml. Plasma histamine was significantly elevated by infusion of histamine at 0.05 to 1.0 μg/kg/min or by cold immersion of the hand of a cold-urticaria patient. Therefore this modified isotopic-enzymatic assay of histamine is extremely sensitive, capable of measuring fluctuations in plasma histamine levels within the normal range, and potentially useful in analysis of the role histamine plays in human physiology.