TREM1 regulates antifungal immune responses in invasive pulmonary aspergillosis.

Pattern recognition receptors (PRRs) are responsible for Aspergillus fumigatus recognition by innate immunity and its subsequent immune signaling. The triggering receptor expressed on myeloid cells 1 (TREM1) is a recently characterized pro-inflammatory receptor constitutively expressed on the surface of neutrophils and macrophages. A soluble form (sTREM1) of this protein that can be detected in human body fluids has been identified. Here we investigated the role of TREM1 during invasive pulmonary aspergillosis (IPA). IPA patients displayed significantly higher levels of sTREM1 in bronchoalveolar lavages when compared to control patients. Functional analysis in TREM1 showed that the levels of sTREM1 and TREM1 pathway-related cytokines were influenced by single nucleotide polymorphisms in TREM1. In addition, we confirmed a role of TREM1 on antifungal host defense against A. fumigatus in a murine model of IPA. TREM1 deficiency increased susceptibility to infection in the immunosuppressed murine host. Deletion of TREM1 showed delayed innate and adaptive immune responses and impaired pro-inflammatory cytokine responses. The absence of TREM1 in primary macrophages attenuated the TLR signaling by altering the expression of both receptor and effector proteins that are critical to the response against A. fumigatus. In this study, and for the first time, we demonstrate the key role for the TREM1 receptor pathway during IPA.

Carcinoide de células caliciformes apendicular. A propósito de tres casos / Goblet cells carcinoid of appendix. About three cases

Los carcinoides de células caliciformes del apéndice cecal son una patología infrecuente con características propias dentro de los tumores neuroendocrinos apendiculares. Presentamos tres casos de esta entidad, dos diagnosticados en un cuadro clínico de abdomen agudo y otro como hallazgo incidental en una colectomía derecha por un adenocarcinoma de colon. La media de edad fue de 46,6 años, el 100% fueron mujeres. Los tres casos se trataron con una colectomía derecha laparoscópica. En el seguimiento, una paciente presentó una metástasis del adenocarcinoma de colon en el hígado y otra una carcinomatosis peritoneal, siendo ambas reintervenidas. Dos pacientes están vivas en el momento actual con un seguimiento de 26 meses de media y una falleció a los 16 meses por progresión de su enfermedad. Realizamos una revisión de la clínica, diagnóstico, clasificación y tratamiento de este in-usual tipo de neoplasia

The goblet cells carcinoid of the cecal appendix is an uncommon disease with own characteristics within the appendicular neuroen-docrine tumors. We present three cases of this entity, two diagnosed in a clinical picture of acute abdomen and another as incidental finding in a right colectomy for colon adenocarcinoma. The average age was 46.6 years, 100% were women. All three cases were treated with a laparoscopic right colectomy. In follow-up, a patient presented a metastasis of adenocarcinoma of colon in liver and other a peritoneal carcinomatosis, being both operated. Two patients are alive at the present time with a follow-up of 26 months on average and one died at 16 months due to progression of their disease. We carry out a review of the clinic, diagnosis, classification and treatment of this unusual type of malignancy

Long-lived polyclonal B-cell lines derived from midgestation mouse embryo lymphohematopoietic progenitors reconstitute adult immunodeficient mice.

Lymphohematopoietic progenitors derived from midgestation mouse embryos were established in long-term cultures with stromal cell monolayers and interleukin 7 (IL-7), giving rise to B-lineage cell lines. The initial emergence and in vitro establishment of these early embryo cell lines were highly sensitive to IL-7-mediated signals, in comparison to cell lines similarly obtained using precursors from late fetal liver (> 13 days postcoitum) and adult bone marrow. The early embryo-derived progenitors spontaneously differentiated in vitro to CD19(+)IgM(+) immature B cells in the presence of optimal concentrations of IL-7, in contrast to those progenitors obtained from late gestation and adult mice, whose differentiation only occurred in the absence of IL-7. The newly in vitro-generated B cells of the early embryo cell lines repopulated adult immunodeficient severe combined immunodeficient mice on their adoptive transfer in vivo and generated specific humoral immune responses after immunization.

[Core and cutaneous thermal changes in the upper limb after anesthesia induction]. / Cambios térmicos centrales y cutáneos en el miembro superior después de la inducción anestésica.

OBJECTIVES: To describe and compare the core-cutaneous thermal and photoplethysmographic time-course effects after induction of general anesthesia with propofol, fentanyl and vecuronium. PATIENTS AND METHODS: We measured digital blood flow, core temperature and skin temperature in the upper limb (fingertip, forearm and upper arm) in 20 patients (10 men and 10 women, ASA-I) before anesthetic induction and 5, 10, 15 and 20 min after induction. Skin temperature changes were recorded with disposable thermocouples. Blood flow was recorded by digital photoplethysmography (PhPl) in the thumb. Anesthesia was provided without premedication, using propofol (3 mg.kg-1), fentanyl (0.1 mg) and vecuronium (0.1 mg.kg-1). After endotracheal intubation, anesthesia was maintained with oxygen-nitrous oxide and 0.1 mg of intravenous fentanyl at the tenth minute, without inhaled anesthetics. RESULTS: All patients showed intense, abrupt vasodilatation in the thumb with marked increases in PhPl (PhPl = 10.4 +/- 5.0 mV/V, at 5 min, p < 0.001) and fingertip temperature (TFingertip = 6.2 +/- 2.0 C, at 10 min, p < 0.001). However, skin temperature changes in the upper arm and forearm were moderate and slower (TForearm = 2.1 +/- 1.4 C, p < 0.01 and TUpper arm = 1.1 +/- 1.2 C, p < 0.01; at 20 min in both cases). A significant correlation was found only between PhPl and TFingertip (r = 0.55, p < 0.001). CONCLUSIONS: Anesthetic induction with propofol, fentanyl and vecuronium produces cutaneous vasodilatation in the upper limb unequally: the greatest increase in skin temperature occurs at the fingertip, while forearm and upper arm temperatures increase less. We think that skin vasodilatation in peripheral distal areas may play an important role in redistributing core heat during anesthesia.

Contribution of the outpatient surgery unit ITO the general surgery department of a district hospital.

Introduction: The creation of Outpatient Surgery (OPS) units to combine the quality of medical attention and rationalize costs allows for greater efficiency in the use of resources. Aim: To report our series of patients undergoing surgery at the OPS units integrated into our Hospital (Type II): Patients and method: Between May 1994 and March 1998, 832 outpatients, of a total of 5230, underwent surgery at our General Surgery Unit. The criteria for exclusion from the programme depended on the patient and the enviroment or resulted from the operation itself. Results: Mean patient age was 47.5 years; there were 420 males and 412 females. Surgery was performed for 229 inguinofemoral hernias, 47 umbilical-epigastric hernias, nine incisional hernias, 193 pilonidal sinuses, 156 mammary nodules, 65 varicose veins, 64 arteriovenous fistulae and 69 proctology operations. The most common anesthesia techniques performed were rachianesthesia and local anesthesia. Eight point seven percent of the patients required admission (OPS failure), the most frequent causes being excessive pain, orthostatic-syncopal hypotension, nausea and vomiting and urine retention. There was no morbidity or mortality. Conclusion: OPS is a highly efficient procedure for resolving the most common pathologies in General Surgery. The anesthesia technique was an important factor in the rate of failure.

A regulatory role for Fc gamma receptors (CD16 and CD32) in hematopoiesis.

Progenitor cells of the T- and B-lineages in mice express (CD32) and Fc gamma RIII (CD16) but as the developing lymphocytes begin to express clonal antigen receptors, CD16 and CD32 are downregulated in T-cells, and CD16 is downregulated in B-cells. Considering that counter-receptors for Fc gamma R occur on thymic and bone marrow stromal cells, the possibility exists that Fc gamma R might participate in some aspect of T- and B-lineage development prior to the stage of antigen receptor expression. Previous studies provided evidence that Fc gamma R can influence murine T-lineage development. In the present studies we found that anti-Fc gamma RII/III mAb accelerated B-lineage development in bone marrow cultures from normal mice, but not in cultures from CD16-/- or CD32-/- mice. Similar results were observed when FACS-purified B-progenitor cells were co-cultured with BMS2, a bone marrow stromal cell line. Fresh bone marrow from CD32-/- mice contained about two-fold more B-lineage cells compared to bone marrow from normal or CD16-/- mice. These studies indicate that the Fc gamma R on B-lineage progenitor cells can influence their further development and add to a growing body of evidence that implicates Fc gamma R as regulatory elements in hematopoiesis.

Pancreaticoduodenectomy due to cancer in the elderly.

The elderly have traditionally been excluded from pancreaticoduodenectomy due to the high morbimortality of this procedure. Six cases of pancreaticoduodenectomy) 5 cephalic and 1 total) for periampullary tumors in patients over 70 are reported. There was no mortality. We conclude that, in selected cases, pancreaticoduodenectomy can be performed safely in the elderly.

Homing of transgenic gammadelta T cells into murine vaginal epithelium.

The vaginal epithelium of normal mice contains lymphocytes of fetal thymic origin that express an invariant Vgamma4/Vdelta1 TCR. The apparent lack of other gammadelta TCR species suggests that a selection mechanism might operate to regulate the localization of gammadelta T cells at this anatomical site. Selection might be connected to the Vgamma4/Vdelta1 TCR or to some homing characteristic of the fetal thymic lineage that appears at day 17-18 of embryonic life. In the present studies, we investigated whether transgenic gammadelta cells expressing a TCR species characteristic of the subpopulation of gammadelta T cells found in the blood, spleen and lymph would translocate to the vaginal epithelium. We found that the transgenic Vgamma2 TCR+ cells did accumulate in the vagina of transgenic mice. Furthermore, like normal vaginal gammadelta T cells, the transgenic vaginal gammadelta T cells expressed the phenotype of recently activated memory/effector T cells (CD44(hi), CD62L-, CD45RB(lo), CD69+). Vaginal gammadelta T cells in normal mice do not express the CD2 and CD28 antigens, but both of these markers are present on transgenic vaginal gammadelta T cells. We observed that a small fraction of splenic transgenic gammadelta T cells had the same surface phenotype as the vaginal transgenic gammadelta T cells, raising the possibility that the gammadelta T cells present in the vaginal epithelium of transgenic mice originated from the peripheral lymphoid organs. Data in support of this possibility came from experiments in which co-incubation of splenic transgenic gammadelta T cells with vaginal epithelial cell suspensions induced the vaginal gammadelta phenotype on the splenic gammadelta T cells. The finding of transgenic gammadelta T cells in the vaginal epithelium suggests that homing of gammadelta T cells to this site is not restricted to gammadelta T cells that express the V4/NS1 invariant TCR. Furthermore, these findings imply that retention of gammadelta T cells in the vaginal epithelium of normal mice is affected by a Vgamma4/Vdelta1-specific mechanism. The finding of a significant level of apoptosis in the transgenic vaginal gammadelta T cells, but not in the normal vaginal gammadelta T cells, could reflect that the mechanism of retention of Vgamma4/Vdelta1 + in the vaginal epithelium involves selective survival at the site.

A regulatory role for Fcgamma receptors CD16 and CD32 in the development of murine B cells.

Early in development, murine B-lineage progenitor cells express two classes of IgG Fc receptors (FcgammaR) designated as FcgammaRII (CD32) and FcgammaRIII (CD16), but mature B lymphocytes only express FcgammaRII (CD32), which functions as an inhibitor of B-cell activation when it is induced to associate with mIgM. The functions of CD16 and CD32 on B-lineage precursor cells have not previously been investigated. To search for FcgammaR functions on developing B-lineage cells, normal murine bone marrow cells were cultured in the presence of 2.4G2, a rat monoclonal antibody that binds to CD16 and CD32, or in the presence of control normal rat IgG, and then the B-lineage compartment was analyzed for effects. Cultures that contained 2.4G2 showed enhanced growth and differentiation of B-lineage cells compared with control cultures. The enhancing effect of 2.4G2 also occurred when fluorescence-activated cell-sorted B-cell precursors (B220(+), sIgM-, HSAhigh, FcgammaR+) from normal bone marrow were cocultured with BMS2, a bone marrow stromal cell line, but not when they were cultured in BMS2-conditioned media. The enhancement of B-lineage development induced by 2.4G2 was CD16-dependent and CD32-dependent, because 2.4G2 did not effect B-lineage growth or differentiation in cultures of bone marrow from mice in which either the gene encoding CD16 or CD32 had been disrupted. Analysis of fresh bone marrow from the CD16 gene-disrupted mice showed normal numbers and distribution of cells within the B-cell compartment, but in CD32 gene-disrupted mice, the B-cell compartment was significantly enlarged. These experiments provide several lines of evidence that the FcgammaR expressed on murine B-cell precursors can influence their growth and differentiation.

Galectin-3 down-regulates IL-5 gene expression on different cell types.

Galectin-3 is an animal lectin, formerly named epsilon-binding protein or Mac-2, which has been described to play an important role in some inflammatory processes by the implication of different cells and the increase in cell adhesion functions through laminin binding activity. In this work we analyzed the role of galectin-3 in the modulation of Th2 cytokines that have an important role in the development of the inflammatory response. We have found that the addition of galectin-3 to human eosinophils, the eosinophilic cell line EoL-3, PBMC, and an Ag-specific T cell line (CD4+) produced a selective inhibition of IL-5 transcription. No inhibitory effect was found on the IL-4 mRNA transcription rate. The inhibitory effect on IL-5 transcription was reversed by incubation with lactose and using specific Ab against galectin-3. Galectin-3 is able to induce inhibition of the IL-5 released in the supernatants from PBMC stimulated with phorbol 12,13-dibutyrate and anti-CD3. Similar results were obtained when a T-specific cell line was stimulated with Ag. Also, EoL-3 stimulated with anti-CD32 produced IL-5 protein, the synthesis of which was partially inhibited by galectin-3. The present results demonstrate that galectin-3 induces a selective down-regulation of IL-5 expression in different cell types, opening important new possibilities in the regulation of the allergic reactions.

Fc gammaRII (CD32) is linked to apoptotic pathways in murine granulocyte precursors and mature eosinophils.

Murine granulocytes and precursors express low-affinity IgG Fc receptors (Fc gammaR). We investigated the effects of FcyR ligation on the development of eosinophils in cultures of normal murine bone marrow. Eosinophilopoiesis was induced by culture of bone marrow cells in the presence of cytokines (granulocyte-macrophage colony-stimulating factor [GM-CSF], interleukin-3 [IL-3], and IL-5). Addition to the cultures of 2.4G2, a rat monoclonal antibody (mAb) that reacts with Fc gammaRII (CD32) and Fc gammaRIII (CD16), induced granulocyte apoptosis within 24 hours. Granulocytes in cultures that contained 2.4G2 showed chromatin condensation, binding of Annexin-V, and fas induction, and by electron microscopy, apoptosis was most commonly observed in cells of the eosinophil lineage. Since murine granulocytes can express both Fc gammaRII (CD32) and Fc gammaRIII (CD16), we investigated the effect of 2.4G2 on cultures of bone marrow obtained from Fc gammaRIII (CD16) gene-disrupted mice and found that the apoptosis induced with 2.4G2 was CD16-independent. Studies with bone marrow cultures from B6MLR-lpr/lpr and C3H/HEJ-gld/gld mice established that the Fc gammaRII (CD32)-triggered apoptosis was fas-fasL-dependent. When mature eosinophils isolated from hepatic granulomas of Schistosoma mansoni-infected mice were cultured in cytokines in the presence of 2.4G2, the eosinophils underwent apoptosis within 24 hours. These findings identify a previously unknown linkage between Fc gammaR on eosinophils and fas-mediated apoptosis, a connection that could be relevant to mechanisms by which eosinophils mediate tissue injury and antibody-dependent cellular cytotoxicity reactions.

Gastric surgery does not increase the risk of developing Barrett's esophagus.

OBJECTIVE: Barrett's esophagus is currently believed to be related to severe and prolonged pathological acid gastroesophageal reflux. However, other factors have been discussed, especially pancreatic biliary reflux. To determine the importance of pancreatic-biliary reflux in the genesis of Barrett's esophagus, we assessed the prevalence of Barrett's esophagus in patients with an intact stomach and in those with previous gastric surgery. METHODS: This is a retrospective study in which 22,236 upper digestive endoscopy reports were reviewed and classified into two groups: intact stomach (n = 21,023) and operated stomach (n = 1,213). In turn, these two groups were divided into five subgroups according to surgical techniques. In each of the groups and subgroups, we calculated the percentage of patients with esophagitis, the percentage of esophagitis patients with Barrett's esophagus, and the percentage of Barrett's esophagus patients with complications. Results were compared by chi2 test. RESULTS: With regard to the prevalence of Barrett's esophagus, we found no significant differences between the study groups. CONCLUSIONS: We conclude that previous gastric surgery does not increase the risk that esophagitis patients will develop Barrett's esophagus.

Autoimmune hepatitis type 2 and hepatitis C virus infection: study of HLA antigens.

BACKGROUND/AIMS: Markers for hepatitis C virus are often detectable in patients suffering chronic hepatitis with liver-kidney microsomal type 1 antibodies. Several authors have suggested that two subsets of those patients can be defined: a) hepatitis C virus negative and b) hepatitis C virus positive. The aim of this work was to further analyze the possible genetic association, HLA class I and II, in these two groups of patients. METHODS: HLA was analyzed in 49 patients. Class I was studied using a standard lymphocytotoxicity test and in class II a reverse hybridization-based test for DRB1 typing and PCR-SSO for DQB1 typing were used. Sixty healthy Spanish subjects and 39 chronic hepatitis C subjects without anti-LKM1 antibodies were used as control groups for the "a" and "b" subsets, respectively. RESULTS: No significant association was found with class I specificities in either group. DQB1 typing showed a very significant increase of DQ2 in the "a" group (93.3% vs. 48%; RR = 15; Pc = 0.0025), and DRB1 typing from the "b" group revealed a high association with DR7 (82.3% vs. 43.6%; RR = 6; Pc = 0.0086). CONCLUSIONS: Our studies revealed a strong association with DQ2 for the "a" group and for the first time an extremely high association with DR7 antigen for the "b" subset. Hence it is possible to establish a different genetic profile in these two patient groups.

Lack of Fc-epsilon receptors on murine eosinophils: implications for the functional significance of elevated IgE and eosinophils in parasitic infections.

Chronic infection with Schistosoma mansoni induces in humans and mice a Th2-dominant immune response in which eosinophils and IgE are conspicuously elevated. Human eosinophils express IgE receptors that participate in an IgE-dependent eosinophil-mediated ADCC reaction against Schistosomula larvae in vitro. To investigate the expression of IgE receptors on murine eosinophils, they were purified (>95% pure by Giemsa-stained cytospin preparations) from liver granulomas of Schistosoma-infected mice. Flow cytometric analysis showed the absence of the low-affinity IgE receptor Fc-epsilon RII (CD23) and Mac-2 and the absence of binding of murine IgE. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of granuloma eosinophil mRNA did not detect transcripts for Fc-epsilon RII or the alpha-chain of the high-affinity IgE receptor Fc-epsilon RI, but did detect transcripts that encode Mac-2 and the low-affinity IgG receptors Fc-gamma RIIb2, Fc-gamma RIII, and the FcR-associated gamma-chain. In vitro stimulation of granuloma eosinophils with interleukin-4 (IL-4) did not induce IgE binding, surface expression of Mac-2, or the transcription of Fc-epsilon receptors (Fc-epsilon RI, Fc-epsilon RII/CD23). To investigate normal murine eosinophils, we cultured normal mouse bone marrow cells with recombinant IL-3, recombinant IL-5, and recombinant granulocyte-macrophage colony-stimulating factor, conditions that promote eosinophil differentiation. Flow cytometric analysis of bone marrow-derived eosinophils failed to detect IgE binding or cell surface expression of Fc-epsilon RII and Mac-2, and RT-PCR analysis of fluorescence-activated cell sorted bone marrow-derived eosinophils failed to detect transcripts that encode Fc-epsilon RI or Fc-epsilon RII. These findings show that, in contrast to human eosinophils, murine eosinophils do not express cell surface receptors that bind IgE. However, because IgG receptors (Fc-gamma RIIb2, Fc-gamma RII) were present on eosinophils purified from granulomas, we investigated whether they might be involved in eosinophil activation. We found that an oxidative burst in eosinophils could be triggered through their IgG receptors.

Cytokine mRNA expression in lung tissue from toxic oil syndrome patients: a TH2 immunological mechanism.

In 1981, an epidemic occurred in Spain, toxic oil syndrome (TOS), in people who consumed rapeseed oil denatured with 2% aniline, and it was one of the largest intoxication epidemics ever recorded. In 1989, a similar disease, eosinophilia-myalgia syndrome (EMS) was reported in the USA and was associated with the ingestion of L-tryptophan. The pathologic findings in TOS showed primary endothelial injury, with cell proliferation and perivascular inflammatory infiltrates. Immunologic mechanisms have presumably been operative in the pathogenesis and perpetuation of TOS. Our previous findings pointed to a T-cell activation during acute phase of the disease. In order to analyze which T-cell subset is involved on TOS, we have developed an mRNA extraction procedure from paraffin-embedded lung tissues in patients with pulmonary involvement. We analyzed mRNA expression from different cytokines (IL-1, IL-2, IL-4, IL-5, IFN-gamma, GM-CSF) and CD25 (interleukin 2 receptor) and CD23 (low affinity IgE receptor), using RT-PCR technique. In lung tissues from these patients a T-cell activation was observed. We found a significant increase in Th1 (P = 0.006) and Th2 (P = 0.003) cytokine profile in TOS patients with respect to controls. The increment in TH2 response with respect to TH1 is significant (P = 0.03) in TOS lung specimens. Non-significant differences were obtained in other cytokines and receptors studied as IL-1, CD25, CD23 and GM-CSF. Data presented in this paper are the first clear evidence that an immunological mechanism is directly implicated in this illness.

Study of apoptosis in human lymphocytes by toxic substances implicated in toxic oil syndrome.

Toxic Oil Syndrome is a multisystemic disease that occurred in epidemic proportions in Spain in 1981 caused by the ingestion of rapeseed oil denatured with aniline. Several data implicate T cells in the pathogenesis of the disease. We evaluated the mechanisms of cytotoxicity in human lymphocytes of TOS-related products: aniline, 3-(N-phenylamino)-1,2-propanediol and its mono- and di-oleyl esters and eosinophilia myalgia-related product such as 3-(phenylamino)-L-alanine, which is chemically similar to 3-(N-phenylamino)-1,2-propanediol, and has been found in manufactured L-tryptophan. Our results show that only di-oleyl ester of 3-(N-phenylamino)-1,2-propanediol induces apoptosis in human lymphocytes, in a concentration and time-dependent way, confirmed by morphology, expression of phosphatidylserine in membrane and analysis of DNA degradation.

Eosinophils transcribe and translate messenger RNA for inducible nitric oxide synthase.

Because of the involvement of nitric oxide (NO) in inflammatory states such as parasitic and hypersensitivity disorders and the fact that eosinophils are one of the cell types implicated, we asked whether eosinophils were able to express mRNA specific to inducible NO synthase (iNOS) and iNOS protein and to secrete nitric oxide. iNOS protein was detected on eosinophil preparations by immunocytochemistry using iNOS mAb. Expression of iNOS protein was also detected by immunoblotting in human purified eosinophils and an eosinophilic leukemia cell line, Eol-3. Nitrite production was detected in the supernatant of human eosinophils and Eol-3 cells cultured for 24 h, and was completely inhibited in the presence of the NOS inhibitor N-methylester-L-arginine. iNOS cDNA was obtained by reverse transcription-PCR. After subcloning, sequencing of the 259-bp fragment from three different human eosinophils cDNAs revealed 97% identity with macrophage/monocyte iNOS. Our studies describe for the first time the presence of iNOS on eosinophil and a putative new role for this cell in inflammatory states such as asthma and parasitic disease.

Allergy to olive pollen: T-cell response from olive allergic patients is restricted by DR7-DQ2 antigens.

BACKGROUND: We have recently described the association between the IgE antibody response to Ole e I (the major antigen from olive tree pollen) and the DR7-DQ2 haplotype in a Spanish population. OBJECTIVE AND METHODS: Due to the linkage disequilibrium between DR7 and DQ2, and thus the difficult distinction between the role of these two antigens in the T-cell activation response, we decided to solve this question by two approaches: 1. The study of another ethnic group, individuals of Arabic origin, with a presumably distinct disequilibrium linkage between DR and DQ antigens. Genomic DNA typing was performed in 46 subjects (allergic and non-allergic) by Restriction Fragment Length Polymorphism (RFLP) and results showed that patients with specific IgE antibodies alpha-Ole e I, were DR7 and/or DQ2. These data show a similar restriction pattern to those previously described for Spanish patients. The phenotypic frequency of DR7 antigen is significantly greater than in the non-allergic population, with a corrected P (Pc) value of 0.03. 2. The analysis of the genetic requirements of Ole e I response, using T-cell lines specific for this antigen. This was first carried out by blocking the proliferative response of these T-cell lines with specific anti-human HLA class II antibodies and then testing the genetic restriction of this response using a panel of histocompatible and histoincompatible Antigen Presenting Cells (APCs). Both experiments corroborate the hypothesis that DR7 and DQ2 are implicated in the recognition of Ole e I.