Search

The inhalation of different substances of plant origin can cause immediate and late onset asthma. The list of these agents responsible for such reactions is continuously increasing. We discuss a patient who developed symptoms of asthma after exposure to Pfaffia paniculata root powder used in the manufacturing of Brazil ginseng capsules. Airway hyperreactivity was confirmed by a positive bronchial challenge to methacholine. Sensitivity to this dust was confirmed by immediate skin test reactivity, a positive bronchial challenge (immediate response), and the presence of specific IgE detected by ELISA technique to an aqueous extract. The bronchial response was inhibited by sodium cromoglycate. Unexposed subjects did not exhibit reactivity to this ginseng extract with any of the tests referred to above. The same study performed with Korean ginseng (Panax ginseng) elicited negative results. This study is the first, to our knowledge, that links ginseng-root dust to occupational asthma.

C57BL/6J mice immunized with devitalized Ehrlich tumor (ET) cells produce high serum levels of IgM antibodies to ET cell-surface carbohydrates that are critical in the observed resistance against this tumor. However, this response is not found in ET-bearing mice at any stage of tumor development. Since previous studies had shown splenic natural suppressor (NS) cells in ET-bearers, their role in such IgM impairment was assessed. Here we show that tumor-bearers' spleen cells (TBSC) are unable to produce IgM in vitro in response to LPS, due to the presence of NS cells. Nevertheless, TBSC do produce IgM antibodies to ET cell-surface carbohydrates in increasing amounts as the tumor progresses. Yet these antibodies are not detected in sera of ET-bearers and are greatly decreased in immunized mice with a growing tumor. Moreover, increasing amounts of circulating carbohydrates, able to absorb most specific IgM, are found in ET-bearing sera associated with a large molecular size structure(s). These carbohydrates are also found in ET cell-culture supernatants and cell-free ascites fluid derived from this tumor, indicating their tumor origin. Taken together, our results indicate that lack of specific IgM antibodies in ET-bearing mice is not due to faulty production, but to in vivo absorption by carbohydrates shed from ET cells in increasing amounts as the tumor progresses. Thus, NS cells are unable to suppress this IgM production in vivo, despite the strong suppressor activity they show for many responses in vitro.

Summary Background Cluster immunotherapy is becoming increasingly used. It allows for a rapid build up phase and the administration of higher doses of allergen in a shorter period of time. Objectives To evaluate the effect of short-term pre-seasonal immunotherapy using a glutaraldeyde-modified allergen vaccine in reducing specific nasal hyperreactivity in nasal challenge tests. Materials and Methods Thirty-three patients were selected. All patients had a positive history of allergic rhinitis and skin tests to grass pollen, although most of them (72.7%) were sensitized to other allergens as well. The study was conducted outside of the pollen season and the patients did not receive any pharmacological treatment during this period of time. Two randomized groups of patients were established; Group A: 22 patients (13 females and nine males) and Group B, 11 control patients (seven females and four males). Patients in Group A received immunotherapy with a vaccine containing 50% of the wild grasses Trisetum paniceum and Dactylis glomerata. All patients underwent titrated nasal provocation tests (NPT) before and after completion of the study (2.3 and 2.8 months for Groups A and B, respectively). The administration schedule consisted of 0.1 and 0.2 mL at day 1, followed by 0.3 and 0.5 mL at day 7, 0.5 mL after 2 weeks followed by 0.5 mL monthly. A single vial was used containing an allergen concentration of 10 000 TU/mL (105 μg of total protein and 24.6 μg of Group 1 plus 5 allergens/mL). A mean of 6.5 injections were administered to Group A patients between NPTs. Results There were no significant differences between both groups at the beginning of the study (P=0.48). At the end, only Group A patients needed significant greater threshold concentrations for a positive NPT than at the beginning (P=0.002). Conclusions A short-term cluster pre-seasonal inmunotherapy with a modified vaccine containing a mixture of grass pollen is effective as determined by an objective measure after only a mean 2.3 months of treatment.

In their comment on the article “The Universal Dynamics of Tumor Growth” by Bru et al. (2003), J. Buceta and J. Galeano imply that the analysis presented is incorrect and lacking in rigor. The article in question shows, using scaling analysis, that 16 in vitro-grown tumor lines and 15 in vivo tumors all have the same growth dynamics. These results support the conclusions made in a previous article by our group (Bru et al., 1998), in which the methods for analyzing circular interfaces were developed. In that article, these methods were used to determine the growth dynamics of four clones of the C6 astrocytoma cell line. The critical exponents describing the spatial and temporal invariances of the interface were determined and found to be completely compatible with the molecular beam epitaxy (MBE) universality class. For later discussion, it is important to note that the contours of the clones clearly grew over the 1400 h cultivation period but that this did not alter the results of the scaling analysis—thus the growth dynamics of the tumors did not change either. The values for the critical exponents of local and global roughness were verified using several methods developed (and later published) for use with interfaces that change in size over time (Bru and Casero, 2003). This latter article compares three methods for measuring the local width of an interface and shows that the local roughness exponent values do not change simply because the system changes in size. In fact, this article analyzes models that show the apparent scaling anomaly to be an effect of the interface analysis methodology. In Bru et al. (2003) (the article commented upon by Buceta and Galeano), we show how the growth dynamics of 15 tumor lines and nontumor cell colonies, and 16 animal and human tumors in vivo, are all completely compatible with MBE universality—strongly suggesting that these dynamics describe the growth of all tumors. Moreover, the spectra provide a coefficient of global roughness of 1.5; the only universality class compatible with this value is MBE (as clearly shown in the figures in the article), as well as the rest of measured critical exponents. In this article (Bru et al., 2003), we clearly state that if all tumors grow with MBE dynamics, then they should also show three classic features of the MBE system. The first of these is a linear growth rate (Bru et al., 1998), something easily shown by averaging the terms of the standard MBE universality equation. The second, which in some ways is an implication of the first, is that any growth will be restricted mainly to the border of the system (Bru et al., 1998). Finally, the third is arrived at by considering the fourth derivative of the MBE universality class equation (which expresses the dominant mechanism of the growth process), and this clearly shows that tumor growth occurs by diffusion of newly produced cells at the tumor border. These three characteristics can all be deduced from the MBE equation, which we believe represents the growth dynamics of all solid tumors. It might be said that the growth dynamics of a system are “written in its interface”. Scaling analysis is one of the most powerful tools ever developed for determining the dynamic component of systems with rough interfaces. Analysis of the spatial and temporal invariances allows the growth dynamics to be determined—and therefore the mechanisms responsible for growth. These invariances are quantified using the values for a set of very robust critical exponents. Scaling analysis thus allows a standard growth equation to be determined, and with this the universality class of the dynamics and the basic growth mechanisms of the system. It is this that most surprises people not familiar with scaling analysis but which so obviously marks the difference with what might be called conventional modeling. The latter tries first to determine the hypothetical mechanism of growth and from this the mathematical equation that governs the process. Scaling analysis allows the reverse. The argument put forward by Buceta and Galeano is, therefore, just the opposite of a scaling analysis approach. They first argue that a system with a constant growth rate does not necessarily mean that MBE dynamics are at work. What they fail to realize is that implications do not always work in both directions. It is absolutely clear that not every system with a linear growth rate has to have MBE dynamics, but every system with MBE dynamics has to have a linear growth rate (this can be obtained theoretically by averaging the terms in the equation). Secondly, Buceta and Galeano criticize the implication that MBE dynamics must mean that growth is restricted to the system's border. But this is precisely what all systems with MBE dynamics—and we know of many—always show. The basic mechanism of growth is most certainly diffusion of cells at the tumor border, as the values of the critical exponents reveal. Not only are there a great many biological reasons why this should be so (including the fact that the cells inside a tumor have no room to grow), we show this to be true experimentally in 16 cell lines and 15 tumors. Not only is this theoretically and experimentally the case, it is the main point we make in our article. If the cells at the border are the only ones that proliferate, then whatever is inhibiting those in the center from growing could provide the basis of an antitumor therapy (Bru et al., 2004; A. Bru, S. Albertos, F. Garcia-Oz, and I. Bru, unpublished). Also, Buceta and Galeano state that several different types of diffusion process exist. This is true, but it is also thoroughly well known that not all are a product of MBE dynamics. What we state in our article is that if tumors grow with MBE dynamics, then there must be a diffusion process; and we provide the experimental evidence that confirms it. Buceta and Galeano take the characteristics mentioned above independently and argue that on their own they do not imply MBE dynamics are at work; this is their premise for believing our work is not sufficiently rigorous and that it might even be incorrect. But we never once say that they do imply MBE dynamics are present, and much less independently. What we say is that the system has MBE dynamics and therefore shows these characteristics. Indeed, a question our critics do not answer is whether these three experimentally confirmed characteristics acting together could be a reflection of anything else. Finally, and equally unjustifiably, Galeano and Buceta argue that the incoherence and the error of our analysis stem from not taking into account the spatial dilation they describe (Galeano et al., 2003). We here remind the reader that our work was experimentally confirmed using the method of Bru and Casero developed for measuring the local roughness of systems with interfaces that change in size (Bru and Casero, 2003). In this earlier article, we argued the well-known fact that in systems in which the interface varies in size, the interface width need not saturate (as Buceta and Galeano quite rightly state). However, the results they present on the growth of calluses cannot be extrapolated to tumor growth. The interfaces of these calluses did not change in size over time, whereas those of our cell colonies did (by several orders of magnitude over the experimental period; if there were any effect influencing scaling, we would certainly have seen it, especially since the length of time over which we cultured our cell lines is rather longer than anything that can be found in the literature). The lack of growth of the calluses in Bruceta and Galeano's work is clearly reflected in the fact that their interface spectra (which were very noisy) showed no temporal changes. This explains the collapse they obtained; the spectra did not change over time, so a collapse was inevitable from the outset with the spatial dilation factor they propose (in fact the results would have been the same for many other spatial dilation factors). Thus, they specify no growth rate because there is none to give (the contours at different times they provide all fall within the spatial discrimination). In addition, all the calluses developed similarly (almost within the spatial discrimination) and curiously with the same fractal dimension (which appears not to change over time). These authors even make the claim that the same fractal dimension should occur in all plant species. Nevertheless, the analysis presented in that article has nothing to do with tumor growth, not because of the methodology, which is the same as that used in (Bru et al., 1998), but because of the nature of the system studied and the biological interpretations that can be made (in fact Galeano and Buceta fail to make any biological conclusions). Indeed, the dilation proposed by these authors has never been experimentally validated in any other known dynamic system. In finishing, we would like to point out that the characteristics of MBE dynamics discussed in Bru et al. (2003, 1998) have not only been rigorously demonstrated but have served as the basis for a successful antitumor therapy currently under development (Bru et al., 2004; A. Bru, S. Albertos, F. Garcia-Oz, and I. Bru, unpublished).

their hardness and dust-generat ing capacity taust be studied. W e think that onion seeds can be considered as occupat iona! allergens. Having proven that bo th in vivo and in vitro methods are useful tools for the diagnosis of this IgE-media ted disease, we expect to find new hypersensitivities to this al lergen among workers in the same factory and to have informat ion about the prevalence of sensitization. Othe r cases may be found elsewhere.

Tumor growth is associated with neutrophilia, thrombocytosis, and extramedullar hematopoiesis. The mechanism(s) accounting for these phenomena is unclear, although granulocyte-macrophage colony-stimulating factor (GM-CSF) and/or granulocyte colony-stimulating factor (G-CSF) released by tumor cells have been involved. We studied whether CSF released by Ehrlich tumor (ET) may play a role. A comparative study was performed with two cell variants (ET and ET/0) growing in euthymic, nude, and SCID mice. Extramedullar hematopoiesis was assessed in the spleen by scoring organ enlargement, wheat germ agglutinin ve+ cells, and interleukin 3-dependent granulocyte-macrophage colony-forming unit (GM-CFU). Both cell lines showed the same cytokine profile by reverse transcriptase polymerase chain reaction, including GM-CSF, G-CSF, and macrophage colony-stimulating factor (M-CSF); yet, only ET cells produced detectable colony-stimulating activity in vitro, mainly due to GM-CSF. No differences in tumorigenicity were noted between ET and ET/0 cells inoculated to normal or immunodeficient mice. An increase in extramedullar hematopoiesis, accompanied by neutrophilia and thrombocytosis, was associated with tumor progression irrespective of the cell line. A strong correlation was obtained between the increase in splenic GM-CFU and tumor mass (r = 0.96, p < 0.0001) that was independent on the tumor cell line, strain of mice, or stage of tumor development. The results point against CSF released by tumor cells and/or reactive host T cells as the only factors involved in the extramedullar hematopoiesis in this tumor model. The remarkable correlation between splenic GM-CFU and the tumor mass still suggests that a factor(s) of tumor origin may play a critical role.

Sialoadhesin (sheep erythrocyte receptor, SER) is a macrophage-restricted adhesion molecule that binds certain sialylated ligands. It is borne by bone marrow stromal macrophages, promoting the interaction with developing myeloid cells, and by a subset of tissue macrophages involved in antigen presentation and activation of tumor-reactive T cells. The expression of sialoadhesin on SER+ macrophages is not constitutive but requires the continuous supply of a sialoadhesin-inducing serum factor. Tumor growth is often associated with marked alterations of myelopoiesis and impairment of T cell activation; yet the expression of sialoadhesin in macrophages derived from tumor bearers has not been addressed. The aim of this study was to assess whether Ehrlich tumor (ET) – a murine mammary carcinoma – growth may modify the sialoadhesin expression by bone marrow macrophages and/or sialoadhesin-inducing activity in ET-bearing sera. Moreover, putative functional sialoadhesin inhibitors produced by ET cells were tested. The results indicate that bone marrow cells from ET bearers show a seven- to eight-fold decrease in SER+ cells as detected by flow cytometry. This is accompanied by an overall decrease in sheep erythrocyte binding to tumor-bearer-derived bone marrow cells, but also by lower numbers of plastic-adherent cells. Functional sialoadhesin expression is preserved at the single-cell level and no inhibitors are found in ET-bearing sera or ET cell culture supernatants. Tumor progression does not impair the sialoadhesin-inducing activity of ET-bearing sera, or the ability of SER− macrophages (e.g. peritoneal macrophages) to respond to such an induction. In conclusion, while SER+ macrophages are greatly decreased in bone marrow from ET bearers, this is not due to a down-regulation of sialoadhesin expression, nor to an impairment of sialoadhesin-inducing factor or to the presence of sialoadhesin-binding moieties of tumor origin, but, more likely, to a decrease of fully mature macrophages.

e13085 Background: PDM08 is a new active compound with antitumor activity in different cancer models. It is a synthetic derivative of the pyroglutamic acid. The lack of direct effect on tumour cells and antitumor activity in immunodeficient models indicates an immune response mediated mechanism. Toxicology and pharmacology preclinical studies showed a favourable safety profile, confirmed by a single-dose placebo controlled healthy volunteer phase I trial. Methods: A single institution open label phase I study was designed (NCT01380249). PDM08 was administered twice a week for four-week cycles. Initially an accelerate escalation phase was chosen with cohorts of 2 patients who after completing the safety period (2 weeks after last dose) had the possibility of escalate to the next dose level if no grade ≥ 2 toxicity and no progressive disease was observed. After 4 cohorts without meaningful toxicity, this design was amended to open new dose levels after a two-week period of treatment completed by the last patient of the cohort if no grade ≥ 2 toxicity was found. The trial was approved by the Ethics Committee and the Spanish Drug Agency. Extensive pharmacokinetic (PK) and pharmacodynamic (PD) data including serum cytokines, lymphoid subpopulations and immunoglobulins are collected. Preliminary efficacy was evaluated by PET-CT. Results: Since July 2011, 10 patients (pts) were recruited into 6 different dose levels (2pts were escalated) ranged from 0.560 mg to 14 mg. Pt characteristics were: female-male ratio 1:1. Median age: 67 (47-76). ECOG 0: 6 pts. 8 pts had colorectal cancer, 1 lung adenocarcinoma and 1 ovarian carcinoma. Median of previous treatments was 3 ranged 2 to 5. 3 pts presented G1 headache. No Dose Limiting Toxicities were found. Preliminary outcome data showed: best response stable disease for 4 patients, 2 of them being stable for 12 weeks. After 40 target lesions evaluated by PET-CT 6 (15%) showed a reduction in the maximum Standard Uptake Value (SUV) of 18-FDG. Conclusions: PDM08 is the first compound of a new class of drugs that can be dose-escalated safely in an accelerated manner and has promising activity. Last patient will be included in March 2012 so final data for toxicity, PK, PD and efficacy will be provided at the meeting.

Background D. pteronyssinus and B. tropicalis are the most common house dust mite species worldwide. D. pteronyssinus and B. tropicalis are the most prevalent species in the tropics. Twelve allergens have been described in B. tropicalis and fifteen in D. pteronyssinus. The objectives of this study were: 1) to analyze the prevalence of sensitization to B. tropicalis and D. pteronyssinus on the tropical Island of Martinique; 2) to evaluate the allergenic crossreactivity between D. pteronyssinus and B. tropicalis and 3) to identify common, and species specific allergens, using mass spectrometry (MS). The serum of patients sensitized to a single, or to both species, was further analyzed by western blots. Methods The sera of 1,243 consecutive patients evaluated for allergic rhinitis and/or asthma were evaluated for specific IgE using direct ELISA. All patients resided on the tropical island of Martinique. Crossreactivity between both species was analyzed by ELISA Inhibitions. Proteins of both species were digested with trypsin and analyzed by MS/MS (MALDI TOF/TOF). Different databases were used for the identification of the proteins. Results Sensitization to B. tropicalis was detected in 868 sera and to D. pteronyssinus in 919 sera; 135 were sensitized exclusively to B. tropicalis and 177 to D. pteronyssinus. Specific IgE levels to D. pteronyssinus were significantly higher than to B. tropicalis (Mean 0.67 OD ± 0.89 vs. 0.86 ± 1.04; P < 0.001). The correlation coefficient between specific IgE levels to both species was 0.135. ELISA Inhibition studies demonstrated minimal crossreactivity between both species. Western blots done with the serum of poly or monosensitized patients revealed specific IgE binding to similar proteins. Proteomic analysis revealed the presence of the following allergens: B. tropicalis: groups 2, 3, 4, 6, 9, 14 and groups 1, 2, 3, 4, 8, 11, 14, 15, 16 and 18 in D. pteronyssinus. Species specific, as well as common proteins were detected. Conclusions There is limited crossreactivity between B. tropicalis and D. pteronyssinus. Sensitization to their allergens seems to be a parallel phenomenon. Allergen extracts of both species are needed for the correct diagnosis and treatment of mite allergic individuals in the Caribbean.

Background Allergoids have been successfully used in the treatment of respiratory allergic diseases. They are modified allergen extracts that allow the administration of high allergen doses, due to their reduced IgE binding capacity.They maintain allergen-specific T-cell recognition. Since they are native allergen extracts that have been polymerized with glutaraldehyde, identification of the allergenic molecules requires more complicated methods. The aim of the study was to determine the qualitative composition of different polymerized extracts and investigate the presence of defined allergenic molecules using Mass spectrometry. Methods Proteomic analysis was carried out at the Proteomics Facility of the Hospital Nacional de Parapléjicos (Toledo, Spain). After reduction and alkylation, proteins were digested with trypsin and the resulting peptides were cleaned using C18 SpinTips Sample Prep Kit; peptides were separated on an Ultimate nano-LC system using a Monolithic C18 column in combination with a precolumn for salt removal. Fractionation of the peptides was performed with a Probot microfraction collector and MS and MS/MS analysis of offline spotted peptide samples were performed using the Applied Biosystems 4800 plus MALDI TOF/TOF Analyzer mass spectrometer. ProteinPilot Software V 2.0.1 and the Paragon algorithm were used for the identification of the proteins. Each MS/MS spectrum was searched against the SwissProt 2010_10 database, Uniprot-Viridiplantae database and Uniprot_Betula database. Results Analysis of the peptides revealed the presence of native allergens in the polymerized extracts: Der p 1, Der p 2, Der p 3, Der p 8 and Der p 11 in D. pteronyssinus; Bet v 2, Bet v 6, Bet v 7 and several Bet v 1 isoforms in B. verrucosa and Phl p 1, Phl p 3, Phl p 5, Phl p 11 and Phl p 12 in P. pratense allergoids. In all cases, potential allergenic proteins were also identified, including ubiquitin, actin, Eenolase, fructose-bisphosphate aldolase, luminal-binding protein (Heat shock protein 70), calmodulin, among others. Conclusions The characterization of the allergenic composition of allergoids is possible using MS/MS analysis. The analysis confirms the presence of native allergens in the allergoids. Mayor allergens are preserved during polymerization.

Summary This work describes three patients suffering from bronchial asthma after being naturally exposed to airborne plane-tree pollen. The three patients gave immediate response in skin tests and dual response in bronchial provocations using Platanus hybrida extract. There was specific seric IgE activity against this/these antigen(s) with the CAP system. The three patients also showed significant correlation (P < 0·001) between their rhinitis and asthma symptom-scores registered on their diary cards and plane-tree pollen counts, collected using the Burkard spore trap. Among 187 patients living in Madrid and who came to our centre with a history of rhinitis and/or seasonal asthma, we found a prevalence of positive skin-prick tests to Platanus of 56%, only surpassed by gramineous pollen (Dactylis glomerata and/or Trisetum paniceum) 92% and Olea europaea 63%. The aerobiological sampling of the pollen content of the air in Madrid, carried out between 1 January 1979 and 31 December 1993 revealed an airborne presence (per cent of total yearly pollen count, mean of 1979–1993) of 14·9% for the Platanus, 14·8% for grasses, 9·8% for Olea and 3·6% for Plantago. The Platanus is one of the most frequently found pollens in the atmosphere over Madrid. At present, in this geographical area, a high percentage of patients with pollinosis are sensitive to this pollen. At least in some of these patients Platanus pollen is capable of inducing rhinitis and bronchial asthma.

CASE REPORTS Patient 1 is a nonsmoking, 45-year-old homemaker. For years she had been experiencing episodes of rhinoconjunctivitis, dyspnea, and coughing when chopping onions. In the last few years, eczematous dermatitis had been appearing, mainly on the fingertips, which became worse when she was chopping onions. Total serum IgE was 182 IU/ml. Patient 2 is a 37-year-old female office worker and homemaker in whom rhinoconjunctivitis and bronchial asthma caused by sensitivity to pollen were diagnosed. Over the past 15 years, she had noticed intense rhinoconjunctival symptoms and slight dyspnea when chopping onions. Total serum IgE was 162 IUlml. In order to study the prevalence of allergy to onion, skin prick tests were performed on 106 subjects, randomly chosen in our clinic: 39 subjects were atopic. In eight (7.5%) immediate positive reactions were noted. Of these eight, all were sensitive to grass pollen; two of them (patients 3 and 4) had intense rhinoconjunctivitis, and one of the two also had chest tightness, wheezing, and dyspnea after being exposed to the aerosols emanating from onions. Patient 3 was a 31-year-old, nonsmoking man, in whom pollen-induced rhinoconjunctivitis and asthma

We performed an aerobiologic observation of the grasses present in Madrid for 14 years (1978-1991), using volumetric air samplers. The counts obtained show that the major grass pollen release period (average daily grass pollen counts greater than 50 grains/m3 of air) occurs in the months of May and June, although lower counts can occur some days from the end of January onward. There are wide year-to-year variations in total atmospheric grass pollen counts, expressed as the total sum of the mean daily concentrations from April 1st to July 30th (ranging from 2568 to 6624). A strong, statistically significant correlation, based on Spearman's rank test and/or simple and multiple linear regressions, was found between the total grass seasonal count and preseasonal rainfall from October to March (R2 = 0.64; P = 0.0429). The meteorological variable which gave the correlation with greatest statistical significance (R2 = 0.97; P = 0.0016) was the average monthly preseasonal humidity from October to March. A good correlation was also found between March estimates of wheat, rye and barley crops and the total grass count (R2 = 0.73; P = 0.006). A model was designed from the above mentioned humidity variable through a multilinear regression analysis, and it was possible to predict, at the beginning of April, total seasonal counts for 1989 (predicted = 5468; actual = 4410; average error = 24%), 1990 (5033; 6090; -17%) and 1991 (3930; 2568; 53%). These data may help clinicians to predict and prepare themselves for the intensity of the grass pollen season and to explain yearly variations in the severity of symptoms.

We describe the presence of IgG antibodies reacting with histones previously treated with proteases in a patient with vasculitis. The patient's serum did not react with non-treated histones and when several enzymes were tested separately, only α-chymotrypsin reproduced the effect. Reactivity was directed against histone fraction H2B and no other autoantibody was found in the patient's serum. This could represent an autoantigendriven response, histones hydrolyzed in vivo with proteases being the immunogenic stimulus. Diagnostic and pathogenic implications derived from the existence of such autoantibodies are discussed.

Changes in concentrations of IgM and IgG antibodies to Brucella were monitored for at least 13 mo by an enzyme-linked immunosorbent assay (ELISA) in 52 patients with culture-positive brucellosis. Two main patterns were observed. After an initial peak, 29 patients (56%) had a steady drop in their IgG levels, whereas 23 (44%) had more than one peak over time. All patients with a chronic form of brucellosis or a relapse were in the second group. In most cases, Brucella antibodies, although falling to low levels, remained measurable. Cutoff levels for IgM and IgG were calculated after considering serum antibody concentrations in people who had recovered from an infection. A separate normal range was established for occupationally exposed workers. On admission, sera from all patients contained Brucella antibody levels greater than established cutoff levels. Our results show that ELISA is an excellent method for diagnosis and follow-up of brucellosis.

Spleen cells from C57BL/6J mice bearing Ehrlich carcinoma growing as a solid tumor show progressive unresponsiveness to concanavalin A (Con A) and lipopolysaccharide (LPS) mitogens. This is accompanied by striking spleen enlargement with marked hematopoietic activity. Lymphoproliferative assays of normal spleen cells in co-culture with tumor-bearing spleen cells (TBSC) show that: (a) TBSC contain non-specific suppressor cells able to abrogate both Con A and LPS responses, or mixed lymphocyte reaction, of normal spleen cells and (b) suppression by TBSC is MHC-unrestricted, non-prostaglandin-mediated and greatly enhanced by Con A su-pernatants. Suppressor cells associated with TBSC are large, low-density cells without markers of mature B or T lymphocytes or of the mononuclear phagocyte system. Most appear to be asialo-GM,-negative, as suppression was only partially inhibited by treatment with anti-asialo-GM, and complement. Since NK activity is lacking in TBSC, our data strongly suggest that these “null” suppressor cells are related to the natural suppressor (NS) cells found described in normal bonemarrow and neonatal spleens, or induced in adult spleens by total lymphoid irradiation, graft-vs.-host disease, or cyclophosphamide treatment.

We report a case of an 8-year-old atopic boy in whom ingestion of a chamomile-tea infusion precipitated a severe anaphylactic reaction. The patient suffers from hay fever and bronchial asthma caused by a variety of pollens (grass, olive, and mugwort). This severe reaction was developed after his first ingestion of chamomile tea. Studies revealed the presence of immediate skin test reactivity and a positive passive transfer test to chamomile-tea extract. Moreover, both specific antichamomile-tea extract and anti-Matricaria chamomilla-pollen extract IgE antibodies were detected by an ELISA technique. Cross-reactivity among chamomile-tea extract and the pollens of Matricaria chamomilla, Ambrosia trifida (giant ragweed), and Artemisia vulgaris (mugwort), was demonstrated by an ELISA-inhibition study. These findings suggest a type I IgE-mediated immunologic mechanism as being responsible for the patient's anaphylactic symptoms and also suggest that the patient cross-reacted the pollens of Matricaria chamomilla contained in the chamomile tea because he was previously sensitized to Artemisia pollen.