Histamine suppression of in vivo eosinophil accumulation and histamine release in human allergic reactions

Although it has been shown that histamine inhibits antigen-induced in vitro histamine release from basophils, it is unclear whether histamine inhibits in vivo mediator release in human allergic reactions. We report effects of exogenous histamine on histamine release and inflammatory cell responses in antigen-challenged skin sites in eight ragweed-sensitive individuals. Four heat-suction blisters in each subject were unroofed, and a collection chamber was appended to each blister base. Chamber A contained 1000 PNU/ml ragweed extract; chamber B contained buffered saline (control fluid); chamber C contained 1000 PNU/ml ragweed and 50 ng (5 x 10(-7) M) of histamine; and chamber D contained histamine alone (50 ng). Comparative analyses of chamber histamine levels in individual subjects showed that (1) histamine levels in chamber A were significantly greater than those in chamber B (p less than 0.01) and that histamine levels in chamber C were not significantly different than those in chamber D (p less than 0.5). Likewise, comparison of eosinophils attaching to membrane filters appended to the chamber bases for 2 hr showed that there were significantly more eosinophils in chamber A than in chamber B (p less than 0.01) and that there was no significant difference in eosinophil numbers on filters appended to chamber C vs chamber D. In three of four subjects studied, addition of exogenous histamine (50 ng/ml) to ragweed before intradermal injection inhibited the ultrastructural mast cell alterations seen within 10 min after injection of ragweed alone. In the one subject in which mast cell alterations were not prevented, exogenous histamine also did not inhibit antigen-induced histamine release or subsequent eosinophil accumulation in the skin chambers.     

Histologic studies of human skin test responses to ragweed, compound 48-80, and histamine

The patterns of mast cell and eosinophil changes at the site of intradermal injection of ragweed, compound $\text{48}\text{80}$, and histamine were studied by histologic techniques. Dermal biopsies of 23 ragweed skin test-positive subjects revealed increasing numbers of eosinophils and decreasing numbers of mast cells over a 240 minute period following injection of antigen. The eosinophil response began in the periappendigeal areas, extended into the interstitium, and occurred in sites with depressed wheal reactions in hydroxyzine-treated subjects. These cellular patterns did not occur at ragweed skin test sites in nonatopic subjects. However, compound $\text{48}\text{80}$ induced similar eosinophilic responses in both atopics and nonatopics. Histamine injection was followed by no eosinophilic or mast cell changes. These findings support the hypothesis that eosinophil responses in reagin-mediated reactions may occur secondary to release of a mediator other than histamine of mast cell origin.     

Histologic studies of human skin test responses to ragweed and compound 4880: II. Effects of corticosteroid therapy

In a controlled study, a 1-week course of a moderate dosage of corticosteroids given to ragweed-sensitive subjects was associated with a statistically significant decrease in blood eosinophil levels and tissue eosinophil, but not mast cell, responses to ragweed antigen and compound $\text{48}\text{80}$, This therapy did not affect IgE-class anti-ragweed antibody levels or gross whealing responses to antigen or compound $\text{48}\text{80}$. While these findings suggest indirectly that a major component of the corticosteroid effect is likely an alteration of the mobilisation of eosinophils to the inflammatory site, additional studies will be required to more directly characterize the mechanisms involved.     

Histologic studies of human skin test responses to ragweed and compound 48/80: III. Effects of alternate-day steroid therapy

Our previous studies have shown depressed eosinophil responses in skin test reactions to pollen antigens and compound 48/80 in those just completing a 1-wk course of daily steroids. Wheal reactions were unaffected. In this study, 6 ragweed-sensitive atopic subjects were studied before and on the seventh day (“day on”) and day 8 (“day off”) of a course of alternate-day steroids. Blood neutrophil levels rose on day 7 and were similar to baseline on day 8, whereas blood eosinophil levels were significantly reduced on both days 7 and 8. Neutrophil responses in skin test reactions were depressed on day 7 and normal on day 8. In contrast, the tissue eosinophil responses were depressed significantly, and to similar degree, on both days 7 and 8. These findings are of potential significance in evaluating the clinical effects of steroids in allergic diseases.     

Histamine release and complement changes following injection of contrast media in humans.

Mechanisms responsible for allergic-like reactions following administration of radiographic contrast media (RCM) are unclear. Aortic root blood specimens were obtained sequentially in 6 subjects following injection of RCM into the pulmonary artery during cardiac catheterization. In 5 subjects, elevated plasma histamine levels (up to 80 ng/ml) occurred within minutes. Levels of C3, C4, factor B, and total hemolytic complement activity were decreased in the same specimens. No hemodynamic or clinical abnormalities were noted. These findings support the concept that RCM can liberate histamine in vivo in humans. Complement alterations may be related to localized RCM-protein interaction. It is unclear whether complement changes are related to the RCM-induced allergic mediator release.     

A controlled study of the effect of corticosteroids on immediate skin test reactivity

Limited uncontrolled studies in the past have yielded conflicting results as to the ability of corticosteroids to suppress the immediate wheal-and-flare test. This double-blind controlled study, using 15 atopic volunteers, demonstrated that a one-week course of steroids exerted no significant effect, compared to placebo, on reactivity to ragweed wheal size and the threshold dilution measurements. Reactivity to compound 48–80 and histamine was also unaffected. In vivo effects of the steroid in these subjects was demonstrated by significant eosinopenia.     

Histologic responses in human skin test reactions to ragweed. IV. Effects of a single intravenous injection of steroids.

A controlled study has been carried out dealing with the early effects of a single intravenous dose of either methylprednisolone or placebo or newly developed and ongoing cellular inflammatory responses in immediate hypersensitivity skin test reactions. There was no significant difference in the tissue eosinophil responses to ragweed injected 2 hr before and 2 hr after placebo; there was a significant rise (101% +/- 39) from the second to fourth hour after antigen injection. By contrast, there was a marked decrease in the tissue eosinophil response to antigen injected 2 hr after steroids as compared to the pattern seen in the presteroid reaction. In addition, the eosinophil numbers not only did not increase from the second to fourth hour when steroids were injected at the second hour but decreased markedly. These findings suggest early suppressive effects on tissue eosinophil responses within 2 hr after steroid were administered intravenously. Also, there may be trafficking of eosinophils both into and out of these inflammatory sites during the first hours after intradermal antigen injection.     

The direct demonstration of histamine release in allergic reactions in the skin using a skin chamber technique.

We have adapted a skin chamber technique to permit sampling of fluid at the skin window sites of pollen antigen-induced allergic reactions. Low background levels of histamines are formed in control chambers, whereas significantly increased (p less than 0.01) amounts are found within 30 min following ragweed application in sensitized subjects.     

Effects of theophylline,terbutaline, and prednisone on antigen-induced bronchospasm and mediator release

Eleven subjects demonstrating clinical, skin, and inhalation sensitivity to grass or ragweed pollen underwent serial inhalation challenges, with and without orally administered theophylline, terbutaline, and prednisone. Comparisons of antigen sensitivity and mediator release were made during these challenges. All three drugs significantly reduced antigen sensitivity (PD₂₀ inhalation units increasing from 670 to ≧ 3,280). Peak plasma histamine levels after antigen challenge decreased from 11.4 ng/ml to ≦ 3.4 ng/ml during all drug administrations. Similarly, the percent increase in serum neutrophil chemotactic activity (NCA) also decreased, from 96% to ≦ 36% during drug administrations. However, even at antigen doses resulting in bronchospasm during drug administration the systemic appearance of NCA and histamine were reduced. We conclude that prednisone, theophylline, and terbutaline significantly reduce antigen-induced bronchospasm and mediator release. The occurrence of bronchospasm despite the inhibition of histamine and NCA suggests either that the local concentration of these mediators are critical or that other mediators produce the bronchospasm observed.     

Further characterization and biologic activity of ragweed antigen--induced neutrophil chemotactic activity in man.

We have previously described the appearance in serum of increased neutrophil chemotactic activity (NCA) during bronchospasm induced by inhalation of ragweed antigen in ragweed-sensitive subjects. This NCA is non-complement derived, appears within 1 min after antigen inhalation, and is not seen after methacholine-induced bronchospasm. This article describes further characterization of this chemotactic activity and correlation with in vivo leukocytosis. NCA consistently eluted in the void volume (fraction I) after Sephadex G-150 chromatography of patient serum obtained 10 min postchallenge. Fraction I contained 94% of the NCA of postchallenge whole serum. Both postchallenge whole serum and fraction I deactivated neutrophils to autologous chemoattractants and complement-derived chemotactic factors, but not serum-independent chemotactic factors. NCA was chemotactic for neither human nor guinea pig eosinophils, nor for human mononuclear cells. A significant increase of circulating neutrophils was seen only after antigen-induced bronchospasm and correlated with the increase in NCA. Thus, NCA represents another inflammatory mediator of probable mast cell origin that may explain, at least partially, the accumulation of neutrophils observed in the peripheral blood, skin, and bronchial wall after immediate hypersensitivity reactions.     

Comparison of plasma histamine and cyclic nucleotides after antigen and methacholine inhalation in man

Serial determinations of plasma histamine and cyclic nucleotides (adenosine monophosphate [AMP] and guanosine monophosphate [GMP]) were performed after inhalation of antigen and methacholine in four groups of subjects. In the first group, consisting of six antigen-sensitive subjects exhibiting bronchospasm after inhalation of ragweed or grass antigen, plasma histamine was elevated within 2 min and persisted for 30 min after inhalation of antigen. Peak histamine levels were between 18 to 80 ng/ml. In the second group, consisting of four nonatopic subjects, neither bronchospasm nor histamine was observed, despite inhalation of the same or 10-fold increased concentrations of antigen. In the third group, consisting of six subjects (three atopic and three nonatopic) exhibiting bronchospasm after inhalation of 2.5 to 10 mg of methacholine, sustained increases of histamine began at 1 min and persisted for 60 min after inhalation of methacholine. In the fourth group, seven subjects (two atopic, five nonatopic) without demonstrable bronchospasm despite inhalation of 2.5- to 10-fold increased doses of methacholine, no histamine was detected in the plasma at any time after inhalation of methacholine. Serial measurements of cyclic nucleotides showed no consistent changes in serum levels of cyclic AMP or cyclic GMP following inhalation challenge. We conclude that serum levels of histamine but not cyclic nucleotides change during bronchospasm induced by either antigen or methacholine.     

Systemic reaction to radiocontrast media during hysterosalpingography

Individuals with histories of previous anaphylactoid reactions to injected radiocontrast media (RCM) are not generally considered at increased risk when RCM is subsequently administered by a nonvascular route. We have observed a patient who experienced laryngeal edema, generalized angioedema, and bronchospasm after instillation of RCM during hysterosalpingography. We found no published reports of similar reactions, although intravasation of RCM into the circulation occurs commonly during this procedure, as was seen in this patient. Approaches to hysterosalpingography in RCM-reactive patients are suggested.     

Mediator release in local heat urticaria

We described the sixteenth reported case of local heat urticaria, in a 59-yr-old woman with erythema and angioedema upon contact with hot water or outdoor heat exposure. Immersing her hand in 39° to 40° C heated water resulted in an erythematous, angioedematous response sharply demarcated by the line of immersion and was associated with immediate increases in histamine concentration (18 to 135 ng/ml) and high molecular weight neutrophil chemotactic activity (two to five times prechallenge levels) in venous blood draining the challenge site. We suggest that the local heat urticarial response in this woman was a form of physical urticaria associated with release of mast cell-derived mediators, akin to cold and cholinergic urticaria.     

Cytogenetics of human malignant melanoma and premalignant lesions

Chromosome studies were done on direct preparations, early passage cultures, and/or cell lines derived from melanocytic lesions of 17 patients. There were 5 nevi (3 dysplastic); 1 early primary melanoma (radial growth phase); 1 advanced primary melanoma (vertical growth phase) with multiple metastases; and 10 metastatic lesions. The 5 nevi had normal karyotypes, while each of the tumors had a predominantly abnormal karyotype. The early melanoma was pseudodiploid, including a 6p;22 translocation. Ten of the 11 advanced melanomas had one or more aberrations involving chromosome #1, with 9 having deletions or translocations of Ip that involved the proximal segment 1p12→1p22 9 times in 8 lesions. Six advanced tumors had additional material involving 7q, including extra #7s (4 cases) and 7q (2 cases). Nine melanomas, including the early tumor, had alterations in chromosome #6. Three had additional copies of 6p (as iso6p or t6p); the others showed no consistent pattern. In one advanced tumor, the primary lesion and 5 metastases (removed seriatim over an 18-month period) had nearly identical karyotypes, indicating the clonal nature of the neoplasm. The nonrandom cytogenetic changes suggest that genes important in melanoma carcinogenesis are located on the proximal portion of 1p, on 7q, and on chromosome #6. More data on early lesions are needed to identify the relation of these various cytogenetic changes to the different stages of malignant melanoma development.     

Increased antigen-induced local and systemic mediator release in rhinitis subjects with pulmonary symptoms in the pollen season

In order to delineate parameters that might discriminate between allergic subjects who develop R or R-P symptoms during natural antigen exposure, 26 subjects allergic to grass or ragweed pollen were classified into R or R-P groups, and then the antigen sensitivity and degree of in vivo mediator release were compared. Antigen-skin sensitivity was quantitated by dilutional skin-test titration, and bronchial sensitivity was quantitated by the amount of inhaled antigen required to receive the FEV1 by 20%. Mediator release was determined by measuring the amount of histamine that was released into skin chambers during antigen incubation and the rise in plasma histamine and serum NCA during antigen-induced bronchospasm. Compared to the 13 R subjects, the 13 R-P subjects were: (1) more sensitive to antigen by both skin-test and inhalation challenge, (2) responded to inhalation of antigen with a greater fall in FEV1 and a greater rise in serum NCA and plasma histamine, and (3) released more histamine into skin chambers after antigen incubation. Even when R and R-P subjects were matched by comparing only subjects with equal skin sensitivity to antigen, greater increases in serum NCA and plasma histamine occurred after inhalation of antigen in the R-P subjects. These data are consistent with the hypothesis that allergic rhinitis subjects who develop pulmonary symptoms during natural pollen exposure are more sensitive to antigen and release more mediators in response to antigen administration. It is therefore possible that the degree of mediator release may be an important factor in determining the pattern of clinical responses to antigen exposure.     

The proposed origin of double minutes from Homogeneously Staining Region (HSR)-marker chromosomes in human neuroblastoma hybrid cell lines

Double minute chromosomes have been reported in a number of human and animal tumors although their origin and function in the cell remain unclear. The fusion of two cell lines without double minutes—a human neuroblastoma line containing another chromosomal abnormality, the homogeneously staining region (HSR), and a mouse fibroblast line—resulted in hybrid clones containing varying numbers of double minutes. The appearance of double minutes occurred coincident with the disappearance of the HSR. We propose that double minutes can originate from the breakdown of an HSR. The possible functional significance of these two chromosome abnormalities is discussed.     

Antigen-induced neutrophil chemotactic activity in man. Correlation with bronchospasm and inhibition by disodium cromoglycate.

We have previously reported increased neutrophil chemotactic activity in sera obtained after positive antigen inhalation responses in atopic subjects. This report describes the kinetics of appearance of this serum activity and the effects of antigen dose and disodium cromoglycate pretreatment on the response in 10 ragweed-sensitive subjects. Significantly increased chemotactic activity was present as early as 1 min, peaked at 10 min, and persisted through 24 hr after inhalation of antigen. The increased chemotactic activity correlated with the degree of bronchospasm induced by antigen inhalation and the amount of antigen administered. The increased chemotactic activity and bronchospasm were blocked by administration of disodium cromoglycate prior to antigen challenge. These findings are consistent with a postulated antigen-induced anaphylactic release of chemotactic activity. The correlation of this activity with the degree of bronchospasm and its appearance after administration of even small doses of antigen suggest that this activity may be important in antigen-mediated bronchospasm.     

Hypersensitivity reactions to ingested crustacea: clinical evaluation and diagnostic studies in shrimp-sensitive individuals

Adverse reactions to ingested crustacea are common and may be life-threatening. We studied 14 individuals with histories of such reactions to shrimp by immediate skin tests and RAST with extracts of shrimp, crab, crayfish, and lobster. Nine of these subjects (8/8 atopics and 1/6 nonatopics) had positive immediate skin tests (wheal greater than or equal to 2 mm) and RAST (ratios greater than 3.0) to shrimp. Their skin tests and RAST ratios to the other crustacea were also frequently positive even, in several cases, in the absence of prior exposure. In contrast, only 1/10 volunteers with no history of intolerance to crustacea had a weak positive skin test to raw shrimp. These studies suggest that both skin tests and RAST are useful in the confirmation of hypersensitivity to shrimp in atopic individuals and that cross-reactivity among crustacea may exist.