bluuebunny/crossref_metadata_2025 huggingface dataset:
IL‐6 attenuates apoptosis, while neither IL‐6 nor IL‐10 affect the numbers or protease phenotype of fetal liver‐derived human mast cells
Authors: Kambe M. , Kambe N. , Oskeritzian C. A. , Schechter N. , Schwartz L. B.
Published: March 2003

Background The combination of recombinant human stem cell factor (rhSCF), rh interleukin (IL)‐6 and rhIL‐10 was reported to be optimal for mast cell development from cord blood progenitors and to induce chymase expression in all such mast cells earlier in their development than tryptase.

Objective The effects of rhIL‐6 and rhIL‐10 in various combinations on the rhSCF‐dependent development of human mast cells from fetal liver progenitors were examined in serum‐free media.

Methods Dispersed fetal liver cells were cultured in serum‐free AIM‐V medium with rhSCF alone, or with combinations of rhIL‐6 and rhIL‐10. Tryptase and chymase expression, surface Kit expression, metachromasia with toluidine blue and apoptosis were measured.

Results Neither rhIL‐6 nor rhIL‐10 nor the two interleukins together, when included from day 0 of culture, affected the number or protease phenotype of mast cells at 1 or 3 weeks. Expression of tryptase paralleled the appearance of metachromasia and surface Kit, both of which preceded chymase expression, regardless whether a rabbit polyclonal or mouse monoclonal anti‐chymase antibody preparation was used. On the other hand, rhIL‐6 markedly attenuated baseline levels of apoptosis in the presence of rhSCF as well as apoptosis occurring after withdrawal of rhSCF, whereas rhIL‐10 had no effect.

Conclusion RhIL‐6 protected fetal liver‐derived mast cells from apoptosis, particularly after withdrawal of rhSCF, but neither rhIL‐6 nor rhIL‐10 nor the combination of these interleukins affected the numbers or protease phenotype of these mast cells.
An annexin 1 (ANXA1)‐derived peptide inhibits prototype antigen‐driven human T cell Th1 and Th2 responses in vitro
Authors: Kamal A. M. , Smith S. F. , De Silva Wijayasinghe M. , Solito E. , Corrigan C. J.
Published: March 2003

Background Annexin‐1 (ANXA1, lipocortin 1) is a pleiotrophic protein produced by many cell types including peripheral blood leucocytes. Although it has been shown to inhibit ‘macroscopic’ inflammatory processes in animal models, its direct effects on antigen‐activated human T cells have not been studied.

Objective To test the hypothesis that ANXA1‐derived peptides inhibit antigen‐driven prototype Th1 and Th2‐type human T cell responses of clinical relevance and lectin‐driven responses in vitro.

Methods Peripheral blood mononuclear cells (PBMC) were isolated from 14 atopic subjects sensitized to house dust mite allergen (Dermatophagoides pteronyssinus, Der p) and purified protein derivative (PPD) of Mycobacterium tuberculosis. PBMC (1 × 106/mL) were cultured with phytohaemagglutinin (PHA; 5 µg/mL; 4 days), Der p (25 µg/mL; 6 days), PPD (10 µg/mL, 6 days) or medium control. Two ANXA1‐derived peptides, Ac2–26 and AF‐2 (5–500 µm), were assessed for possible inhibition of PHA‐and antigen‐induced T cell proliferation (measured by 3H‐thymidine uptake), while Ac2–26 was assessed for inhibition of Der p‐induced interleukin (IL)−5 release and PPD‐induced interferon‐γ (IFN‐γ) release (measured by ELISA). Comparison was made with dexamethasone as an established inhibitory control. Endogenous production by PBMC of cell surface‐associated and intracellular ANXA1 in response to PHA, Der p and PPD in the presence and absence of dexamethasone was measured by specific ELISA.

Results Both PHA‐ and antigen‐induced T cellular proliferation were inhibited by dexamethasone. Although neither ANXA1‐derived peptide significantly altered PHA‐induced proliferation, both effected concentration‐dependent reductions in antigen‐induced proliferation, Ac2–26 being the more potent. Peptides of identical amino acid composition to Ac2–26 and AF‐2, but of random sequence, were ineffective at equivalent concentrations. In addition, Ac2–26 and dexamethasone inhibited Der p‐induced IL‐5 release and PPD‐induced IFN‐γ release in a concentration‐dependent fashion. Endogenous ANXA1 was detectable in PBMC, but at concentrations approximately 104‐fold lower, in molar terms, than the effective concentrations of the exogenously added, ANXA1‐derived inhibitory peptides. Endogenous production was not significantly altered by any of the T cell stimuli employed in this study, in the presence or absence of dexamethasone.

Conclusion In prototype Th1 and Th2‐type human T cell responses, ANXA1‐derived peptides can inhibit antigen‐driven cellular proliferation and cytokine production.
Levels of cysteinyl leukotriene receptor mRNA in human peripheral leucocytes: significantly higher expression of cysteinyl leukotriene receptor 2 mRNA in eosinophils
Authors: Mita H. , Hasegawa M. , Saito H. , Akiyama K.
Published: March 2003

Background Cysteinyl leukotrienes (CysLTs) have been implicated as important contributors in the pathophysiology of asthma and their biological effects are mediated by at least two distinct G‐protein‐coupled receptors. cDNA sequences of cysteinyl leukotriene receptor 1 (CysLTR1) and cysteinyl leukotriene receptor 2 (CysLTR2) have recently been elucidated.

Objectives Our aim is to explore gene expression and the comparative expression of CysLTR1 mRNA and CysLTR2 mRNA in human peripheral blood leucocytes.

Methods Gene expression of CysLTR1 and CysLTR2 mRNAs in human peripheral blood eosinophils, neutrophils, monocytes and T lymphocytes has been measured by competitive reverse transcription‐polymerase chain reactions using RNA or DNA competitors.

Results (a) When cellular levels of CysLTR1 mRNA were normalized to those of G3PDH mRNA, the relative concentration of CysLTR1 mRNA in eosinophils (43.8 ± 37.2, n = 29) was significantly higher than that in neutrophils (18.7 ± 23.3, n = 11), monocytes (0.93 ± 1.1, n = 10) and T lymphocytes (3.4 ± 2.4, n = 11). (b) When measured using each DNA competitor, mRNAs for both types of CysLTR coexisted in each type of leucocyte. The ratio of CysLTR1 mRNA to CysLTR2 mRNA was significantly lower in eosinophils (0.65 ± 0.42, n = 12) than in neutrophils (6.9 ± 4.9, n = 12), monocytes (1.8 ± 0.9, n = 10) and T lymphocytes (4.5 ± 5.7, n = 10). (c) Human umbilical vein endothelial cells expressed CysLTR2 mRNA, but not CysLTR1 mRNA.

Conclusion These studies reveal that CysLTR1 mRNA and, in particular, CysLTR2 mRNA are abundantly expressed at high levels in eosinophils, raising the possibility that CysLTR2 may have an important physiological role in eosinophils and a CysLTR2 antagonist may be a good target for preventing signal transduction by CysLTs in eosinophils.
Exposure to wheat allergen and fungal α‐amylase in the homes of bakers
Authors: Vissers M. , Doekes G. , Heederik D.
Published: March 2003

Background Few data are available on exposure to occupational allergens in dwellings occupied by inhabitants with occupational exposure to allergens. In small bakeries working and living often takes place in the same building. It is possible that allergens from the bakery can be transported into the homes of the bakers, via the clothes or shoes of the baker.

Objective The aims of this study were to investigate exposure to occupational allergens, wheat and fungal α‐amylase in the homes of bakers, and evaluate potential determinants of exposure. Sensitization in family members to occupational allergens was investigated in a small preliminary survey.

Methods Floor dust samples were collected in the homes of 34 bakers. Levels of wheat and fungal α‐amylase allergens were determined in an extract of the dust samples. Blood samples were collected from bakers and their family members to determine the prevalence of sensitization to occupational allergens.

Results The concentration of wheat and α‐amylase allergens ranged from 38.9 to 172.4 µgeq/m2 (GM), to 10.5–76.7 ngeq/m2 (GM). Higher levels of dust and allergens were measured when the house could be reached directly through the bakery, and in houses with textile floor covers. Higher concentrations were also measured when bakers brought their work clothes and shoes into the house and when textiles from the bakery were laundered at home. Some family members appeared to be sensitized to wheat flour and α‐amylase, but it cannot be excluded that they became sensitized because of their incidental presence in the bakery.

Conclusions Occupational allergens can be found in house dust from the homes of bakers and levels are associated with hygienic behaviour and distance to the bakery.
The Western blot is a highly sensitive and efficient technique in diagnosing allergy to wasp venom
Authors: Zollner T. M. , Spengler K. , Podda M. , Ergezinger K. , Kaufmann R. , Boehncke W.‐H.
Published: March 2003

Background Diagnosis of allergy to wasp venom and decision to perform immunotherapy are based on the patient's history, along with skin and in vitro tests.

Objective Given the high prevalence of specific IgE also in non‐allergic individuals, we evaluated the sensitivity and specificity of Western blots as a possible alternative to serum analyses of venom‐specific IgE.

Methods Skin prick and/or intracutaneous tests were performed in 30 patients with allergy to wasp venom (generalized reaction following sting) along with serum analysis of venom‐specific IgE (AlaSTAT microplate) and Western blots. Western blots were subsequently scanned and evaluated qualitatively and semiquantitatively by means of densitometry. Bands were scored ‘positive’ in cases of signal intensities beyond the mean plus 3 standard deviations of control sera. Twenty newborns (age 2–7 days) and 30 adults without systemic or increased local reactions to hymenoptera stings served as controls.

Results Western blot sensitivity reached 100% in the samples studied and was thus superior to the sensitivities of serum analysis of venom‐specific IgE using AlaSTAT microplate assay (90%) and skin tests (87%). The sensitivity of detection of a phospholipase A1 and antigen 5‐specific band was higher compared with a hyaluronidase‐specific band (97%, 97% and 86%, respectively). Twenty‐four out of twenty‐nine (83%) patients exhibited specific IgE antibodies against at least three distinct allergens. With regard to the specificities, skin tests as well as AlaSTAT microplate assays were comparable (90% and 93%, respectively), whereas the specificity of the Western blots was 70% if the appearance of any single band was regarded as a positive result. However, when analysing the appearance of a specific band for antigen 5 or hyaluronidase the specificity and overall diagnostic value increased markedly, making it the most efficient test (specificity 97% and 100%, efficiency 96.8% and 93.2%, respectively).

Conclusion As allergy to wasp venom is a severe and potentially life threatening disease, false‐negative test results need to be minimized. Therefore, the superiority of the Western blot with regard to sensitivity, specificity and overall efficiency makes this technique a valuable tool for its diagnosis.
Role of conformational and linear epitopes in the achievement of tolerance in cow's milk allergy
Authors: Vila L. , Beyer K. , Järvinen K.‐M. , Chatchatee P. , Bardina L. , Sampson H. A.
Published: March 2003

Background Cow's milk (CM) is one of the leading causes of food allergy in children. However, approximately 85% of milk‐allergic children become clinically tolerant to CM within the first 3 years of life. The mechanisms involved in the achievement of tolerance remain unknown.

Objective To study whether IgE antibodies from children with persistent cow's milk allergy (CMA) differ from children who become clinically tolerant in their ability to recognize linear and conformational epitopes of αs1‐ and β‐casein.

Methods Thirty‐six milk‐allergic children were included in the study: 11 of the children became clinically tolerant, and 25 had persistent CMA. Blood was obtained from all patients during the time they showed clinical reactions to milk challenge. Six non‐milk‐allergic children served as controls. Specific IgE antibodies against linear (denatured) as well as conformational (native) milk proteins were determined by probing dot‐blots with patients' sera. In addition, selected decapeptides from αs1‐ and β‐casein, previously found to be suggestive of persistent CMA, were synthesized on a cellulose‐derivatized membrane and probed with individual sera from 10 patients who outgrew CMA and from 10 patients with persistent CMA.

Results Analysis of immunodot‐blots showed that, in comparison to tolerant patients, milk‐allergic children with persistent symptoms had a significantly higher ratio of specific IgE antibodies to linearized than to native α‐ and β‐casein (P < 0.005 and P < 0.02, respectively). Comparing the selected decapeptides, six of the 10 patients with persistent allergy recognized the peptide corresponding to amino acids 69–78 from αs1‐casein while none of the patients who outgrew CMA had IgE binding to this epitope.

Conclusion Patients with persistent milk allergy possess higher detectable levels of IgE antibodies to linear epitopes from αs1‐ and β‐casein than children who have achieved tolerance. Specific IgE binding to particular linear epitopes in αs1‐casein may be a predictive factor for persistence of CMA.
Safety of rofecoxib in subjects with a history of adverse cutaneous reactions to aspirin and/or non‐steroidal anti‐inflammatory drugs
Authors: Pacor M. L. , Di Lorenzo G. , Biasi D. , Barbagallo M. , Corrocher R.
Published: March 2003

Background Adverse reactions to non‐steroidal anti‐inflammatory drugs (NSAID)s are frequent, and the need to identify a safe alternative drug is a common problem in clinical practice.

Objective To assess the tolerability of rofecoxib, a drug that specifically inhibits COX‐2, in a group of NSAID‐sensitive patients.

Methods One‐hundred and four subjects (29 males and 75 females, mean age 35.6 ± 14.1) were enrolled. All subjects had experienced one or more episode characterized by cutaneous symptoms (erythema, and/or urticaria angioedema) following the assumption of NSAIDs; 92 subjects experienced reactions to only one NSAID (single intolerance: SI) and 12 subjects had reactions to multiple NSAIDs (multiple intolerance: MI). Rofecoxib was challenged at the following dosages: ¼ of 25 mg (6.25 mg), ¼ of 25 mg, and ½ of 25 mg (12.5 mg), at intervals of 1 h if no symptoms had developed with the previous administration, in order to reach a cumulative dose of 25 mg. All subjects underwent two double‐blind, placebo‐controlled challenges in two consecutive days.

Results No reactions against placebo were observed. Similarly, no reactions were observed in all subjects both after the first and after the second challenge to rofecoxib.

Conclusions The present study demonstrated that rofecoxib does not have cross‐reactivity to NSAIDs. Rofecoxib is a safe alternative in subjects with previous adverse cutaneous reaction to NSAIDs.
Intranasal application of purified protein derivative suppresses the initiation but not the exacerbation of allergic rhinitis in mice
Authors: Hattori H. , Okano M. , Yamamoto T. , Yoshino T. , Yamashita Y. , Watanabe T. , Satoskar A. R. , Harn D. A. , Nishizaki K.
Published: March 2003

Background Several epidemiological and experimental studies have demonstrated that exposure to pathogens such as those from the genus Mycobacterium leads to the suppression of allergic sensitization and inflammation. However, little is known as to whether pathogen‐derived soluble antigens have the potential to modulate the pathogenesis of allergic rhinitis.

Objective We sought to determine whether application of purified protein derivative (PPD) from Mycobacterium tuberculosis can suppress the initiation and/or exacerbation of allergic rhinitis using a recently developed murine model.

Methods First, we investigated whether a single intranasal application of PPD could elicit cytokine production in the nose by RT‐PCR. BALB/c mice were repeatedly sensitized with Schistosoma mansoni egg antigen (SEA) intranasally without an adjuvant. PPD was applied through different routes either before or after sensitization. The production of SEA‐specific antibodies, nasal eosinophilia and cytokines by nasal lymphocytes was compared among mice that had or had not received PPD treatment.

Results IFN‐γ, but not IL‐4, was detected in the nasal tissue 12 to 48 h after a single intranasal application of 10 µg PPD. Repeated intranasal application of PPD prior to and during sensitization with SEA significantly inhibited the production of both SEA‐specific IgE/IgG1 and nasal eosinophilia. Moreover, it partially inhibited the production of IL‐4 by nasal lymphocytes in response to SEA. Conversely, this treatment led to a significant increase in IFN‐γ production. On the other hand, PPD applied through the footpad had no effect over the same period. Repeated intranasal application of PPD after sensitization with SEA had no exacerbative effect on allergic inflammation.

Conclusion These results indicate that the local application of PPD, and the subsequent induction of IFN‐γ, inhibits the initiation, but not the exacerbation, of allergic rhinitis in mice. This suggests that pathogen‐derived antigens have potential for use in the prevention and prophylaxis of allergic rhinitis.
Polymorphism of the eosinophil cationic protein‐gene is related to the expression of allergic symptoms
Authors: Jönsson U. ‐B. , Byström J. , Stålenheim G. , Venge P.
Published: March 2003

Background We have found a polymorphism in the ECP (eosinophil cationic protein)‐gene at position 434 according to GenBank accession number NM 002935. This polymorphism would cause the change of the amino acid arginine (base at position 434 is G) at position 97 to threonine (base at position 434 is C).

Objective To investigate the prevalence of the ECP‐polymorphism and to screen for disease associations.

Methods DNA of 209 medical students and 76 asthmatic subjects was analysed. The 434 genotype in the ECP‐gene was detected by cleavage of the amplified DNA sequence with the restriction enzyme PstI and analysis of the cleaved product by agarose gel electrophoresis.

Results The prevalences of the polymorphism in the student population were 53%, 39% and 8% for the 434GG, the 434GC and the 434CC genotype, respectively, with allele frequencies of 72% (434G) and 28% (434C). Subjects reporting allergy had a higher prevalence of the 434G allele than non‐allergic subjects (P = 0.0056). Of the students who were Phadiatop‐positive and had allergic symptoms, 79% had the 434GG genotype, whereas the 434GC and 434CC genotypes were present in 82% of those who did not express allergic symptoms (P < 0.001). Among the 76 patients with asthma, patients with allergic asthma had a significantly higher proportion of 434GG compared with patients with non‐allergic asthma (P = 0.04). None of the 18 subjects of the two groups with the 434CC genotype had allergy.

Conclusion The 434(G > C) polymorphism in the ECP‐gene is related to the development of allergic symptoms, suggesting a central role for the ECP molecule in the process.
Identification of the soybean hull allergens involved in sensitization to soybean dust in a rural population from Argentina and N‐terminal sequence of a major 50 KD allergen
Authors: Codina R. , Ardusso L. , Lockey R. F. , Crisci C. D. , Jaén C. , Bertoya N. H.
Published: March 2003

Background Sensitization to soybean hull (SH) allergens occurs in subjects from Argentina, a soybean producer country. However, the causative allergens have not been identified. The purposes of this study are to: (i) identify the SH allergens using sera of 29 subjects with asthma and/or allergic rhinitis from Argentina exposed to soybean dust who have a positive (weal with SH/weal with histamine ≥ 0.5) skin prick test to SH; and (ii) determine the N‐terminal amino acid sequence of a major 50 K SH allergen that sensitizes this population.

Methods All sera were assayed for specific IgE (RIA), IgG4 (ELISA), and IgE and IgG4‐Western blots. A sera pool from 10 healthy subjects was a negative control. N‐terminal amino acid sequencing was performed by the Edman degradation method.

Results Positive specific IgE only was found in 12/29 (41.4%), IgG4 in 3/29 (10.3%), and both IgE and IgG4 in 14/29 (48.3%) sera. IgE‐Western blot demonstrates: (i) an allergen, MW 50 K (51.7% binding); (ii) one or two distinct allergens, MW < 20.2 K (72.4% binding), depending on the sera; and (iii) 1–5 additional IgE binding proteins, MW > 20.2 to < 46.9 K (41.4% binding), depending on the sera. IgG4‐Western blot demonstrates: (i) a band, MW 70K (31% binding); (ii) a band, MW 50 K (17.2% binding); (iii) one or two additional bands, MW < 20.2 K (51.7% binding), depending on the sera; and (iv) a band, MW > 20.2 to < 28.5 K (20.7% binding). The 50 K allergen N‐terminal amino acid sequence of the first 17 amino acids indicates a significant homology with chlorophyll A‐B binding protein precursors from tomato, spinach, and petunia.

Conclusions Specific IgE and IgG4 to SH are common in sera from allergic individuals living in rural areas in Argentina. SH contain an IgE binding protein, MW about 50 K, not previously described. Sensitization to this allergen is common in subjects who are repeatedly exposed to soybean dust inhalation.
Reproductive history and allergic rhinitis among 31145 Danish women
Authors: Westergaard T. , Begtrup K. , Rostgaard K. , Krause T. G. , Benn C. S. , Melbye M.
Published: March 2003

Background and objective A successful pregnancy is associated with a strong skewing of the immune system towards a Th2‐type immune response. Because such a deviation is also the hallmark of allergic disease, it was investigated whether allergic rhinitis in women was associated with an increased likelihood of becoming pregnant and having a successful outcome of pregnancies.

Material and methods Information on allergic rhinitis and reproductive history was obtained for 31145 pregnant women who participated in a national birth cohort study in Denmark during September 1997 to March 2000, and for whom complete information on siblings and place of residence and birth was available via the Civil Registration System. Data were analysed using logistic regression.

Results Women who had previously been pregnant (OR = 0.91, 95% CI 0.85–0.98) or who had given birth previously (OR = 0.91, 95% CI 0.85–0.98) were less likely to report allergic rhinitis than others. The number of previous spontaneous abortions, gestational week of the first spontaneous abortion and fertility treatment were not associated with allergic rhinitis. Women who had waited less than a year to become pregnant more often had allergic rhinitis (OR = 1.18, 95% CI 1.06–1.32, P = 0.002) than women who had waited for more than a year. Early age at menarche was associated with an increased likelihood of allergic rhinitis (Ptrend = 0.003).

Conclusions Our findings did not support the hypothesis that an atopic genotype overall should be associated with an increased likelihood of successful outcome of pregnancies, but it might be associated with a decreased waiting time to pregnancy.
Long‐lasting effect of sublingual immunotherapy in children with asthma due to house dust mite: a 10‐year prospective study
Authors: Di Rienzo V. , Marcucci F. , Puccinelli P. , Parmiani S. , Frati F. , Sensi L. , Canonica G. W. , Passalacqua G.
Published: March 2003

Background Subcutaneous immunotherapy for respiratory allergy has shown a long‐lasting efficacy after its discontinuation, whereas this evidence is still lacking for sublingual immunotherapy, despite the fact that it is widely used.

Objective We aimed to evaluate whether a long‐lasting effect of SLIT occurs, in a prospective parallel group controlled study.

Methods Sixty children (mean age 8.5 years) suffering from allergic asthma/rhinitis due to mites were subdivided into two matched groups: 35 underwent a 4‐ to 5‐year course of SLIT with standardized extract and 25 received only drug therapy. The patients were evaluated at three time points (baseline, end of SLIT and 4 to 5 years after SLIT discontinuation) regarding presence of asthma, use of anti‐asthma drugs, skin prick tests and specific IgE.

Results We found that in the SLIT group there was a significant difference vs. baseline for the presence of asthma (P ≤ 0.001) and the use of asthma medications (P ≤ 0.01), whereas no difference was observed in the control group. The mean peak expiratory flow result was significantly higher in the active group than in the control group after 10 years. No change was seen as far as new sensitizations were concerned. Specific IgE showed a near‐significant increase (baseline vs. 10 years, P = 0.06) only in the control group.

Conclusion Our study demonstrates that sublingual immunotherapy is effective in children and that it maintains the clinical efficacy for 4 to 5 years after discontinuation.
Epstein–Barr virus-associated peripheral T-cell lymphoma in adults with hydroa vacciniforme-like lesions
Authors: Cho Kwang-Hyun , Kim Chul-Woo , Heo Dae-Seog , Lee Dong-Soon , Choi Won-Woo , Rim Jong-Hyun , Han Won-Suk
Published: March 2003

We describe two Korean adult patients who had necrotizing papulovesicles mainly on their faces. Skin biopsy specimens showed perivascular and periadnexal infiltrate of atypical lymphoid cells with vasculitis in the dermis and subcutaneous tissue. In situ hybridization demonstrated a latent infection of Epstein–Barr virus in the majoriy of lymphoid cells in the dermis. These patients were diagnosed as having T-cell lymphoma. Interestingly, large granular lymphocytosis was found in the peripheral blood of Case 2.
Immunohistology of the early course of lentivirus-induced arthritis
Authors: Von Bodungen U , Lechner F , Pfister H , Vogt H R , Cheevers W P , Bertoni G , Jungi T W , Peterhans E
Published: March 2003

Caprine arthritis encephalitis (CAE) is a lentiviral infection of goats characterized by mononuclear cell infiltration of various tissues, most prominently the joints, mammary glands and, in young animals, the brain. We have investigated the early stages of arthritis induced by intracarpal and intravenous infection with molecularly cloned CAE virus. Analysis of the synovial membranes by immunohistological methods showed that the proportion of CD8+T cells peaked around day 12 post-infection. CD4+ T cells increased to a lesser degree. The relative proportion of B cells rose steadily post-infection. At 33 days post-infection, plasma cells accounted for over one third of all inflammatory cells in the inflamed synovium. Histopathologically, the arthritic lesions in the synovial membranes closely resembled those in membranes of animals with a 2-year history of chronic arthritis. Our observations indicate that this type of short-term experimental infection is particularly suitable for studying the pathogenesis of goat lentiviral infection. In addition, our observations support the view that a predominantly humoral (type 2) immune response may contribute to the pathogenesis of CAE.
Cultured basophils but not cultured mast cells induce human IgE synthesis in B cells after immunologic stimulation
Authors: Yanagihara Y , Kajiwara K , Basaki Y , Ikizawa K , Ebisawa M , RA C , Tachimoto H , Saito H
Published: March 2003

By generating human mast cells and basophils from umbilical cord blood mononuclear cells cultured in the presence of appropriate cytokines, we investigated whether these two cultured cells could provide the cytokine and cell contact signals that are required to induce IgE synthesis in B cells. To activate cultured mast cells and basophils, cross-linking of cell surface high-affinity IgE receptor (FcεRI) was performed with specific antigen after sensitization with murine IgE. Upon FcεRI stimulation, basophils, but not mast cells, secreted significant amounts of immunoreactive IL-4 and IL-13 and expressed detectable CD40 ligand (CD40L) and a very low level of Fas ligand (FasL). These observations at the protein level were consistent with the data obtained at the gene transcriptional level, except for the faint expression of only IL-13 mRNA in mast cells. When added to normal human B cells, activated basophils induced IgE and IgG4 synthesis as well as soluble CD23 release. In contrast, neither IgE nor IgG4 synthesis could be induced by the interaction of B cells with activated mast cells, even in the presence of recombinant IL-4. The induction of IgE synthesis by activated basophils was completely abrogated by two neutralizing MoAbs against IL-4 and IL-13 and by a soluble form of CD40. This abrogation was accompanied by abolished mature Cε transcription in both cases. Addition of anti-FasL MoAb, however, did not significantly affect IgE induction mediated by activated basophils. These results demonstrate that unlike cultured mast cells, cultured basophils produce biologically active IL-4 and IL-13 and express functional CD40L after FcεRI stimulation, thereby contributing to IgE production by B cells, and suggest that relatively weak expression of FasL by cultured basophils is not involved in IgE regulation.
Monocytes modulate enhancement of HIV-1 replication byMycobacterium tuberculosis
Authors: De Haas C J C , De Vos N M , Visser M R , Snippe H , Verhoef J
Published: March 2003

To investigate the effects of Mycobacterium tuberculosis on HIV-1 replication, peripheral blood mononuclear cells (PBMC) of bacille Calmette–Guérin (BCG)-vaccinated donors and non-BCG-vaccinated donors were infected in vitro with a lymphotropic isolate of HIV-1 and cultured in the presence of purified protein derivative (PPD). Addition of PPD resulted in enhanced HIV-1 replication and lymphoproliferation in BCG-vaccinated donor PBMC, while PPD had no such effects in control PBMC. HIV-1 replication increased even more when monocytes were removed from PBMC, while lymphoproliferation was decreased. High percentages of monocytes were associated with a decreased HIV-1 replication and proliferation that could not be reversed by addition of antibodies against the cytokines IL-1, transforming growth factor-beta (TGF-β) or indomethacin. PPD stimulates PBMC to release IL-10, a cytokine known to down-regulate proliferation and HIV-1 replication. PPD-induced effects on proliferation as well as HIV-1 replication could be partially blocked by adding a monoclonal antibody against MHC class II molecules, suggesting that part of the mechanism of PPD-induced enhancement is T memory cell activation.
Modulation of proinflammatory cytokine production in tumour necrosis factor-alpha (TNF-α)-transgenic mice by treatment with cells engineered to secrete IL-4, IL-10 or IL-13
Authors: Bessis N , Chiocchia G , Kollias G , Minty A , Fournier C , Fradelizi D , Boissier M-C
Published: March 2003

TNF-α is one of the major proinflammatory cytokines involved in the pathogenesis of chronic inflammatory joint disease, in human rheumatoid arthritis as well as in murine models of this disease. It was previously described that a highly destructive chronic spontaneous inflammatory arthritis develops in mice expressing a human TNF-α transgene modified with the 3′ untranslated region of β-globin. The present study investigates in this mouse model the effects of the anti-inflammatory cytokines IL-4, IL-10 and IL-13 administered in vivo on proinflammatory cytokine expression. Groups of TNF-α-transgenic mice were engrafted with xenogeneic transfected Chinese hamster ovary (CHO) fibroblasts secreting murine IL-4, IL-10 or IL-13. In vivo treatments consisted of 3 or 4 weekly engraftments, starting when the mice were 4 weeks old. Control groups of transgenic mice were engrafted with β-galactosidase gene-transfected CHO cells or injected with medium. A significant decreased expression of TNF-α transgene, endogenous mouse TNF-α and IL-1 mRNA was observed in splenocytes of mice treated for 3 or 4 weeks with CHO/IL-4 and CHO/IL-13, and, to a lesser extent, with CHO/IL-10, compared with controls. Finally, attenuation of histological scores of arthritides was statistically significant only in the group of CHO/IL-4-treated mice after 3 weeks of treatment (P < 0.05), and was not significant in any other group. These results show that IL-4, IL-10 or IL-13, administered by gene therapy, can decrease the mRNA steady state levels of both endogenous and transgenic cytokines in human TNF-α transgenic mice. In addition, IL-4 can slightly attenuate the development of arthritides in this model.
Modifications of general parameters of immune activation in the sera of Sicilian patients with Boutonneuse fever
Authors: Mansueto S , Vitale G , Mocciaro C , Gambino G , Colletti P , Mansueto P , Spinelli A , Affronti M , Chifari N , Arcoleo F , Milano S , Cillari E
Published: March 2003

The serum levels of β2-microglobulin (β2-M), soluble HLA class I antigen (sHLA-I), soluble CD4 (sCD4) and CD8 (sCD8) were studied in 98 Sicilian patients with Boutonneuse fever (BF). In different stages of infection all markers were significantly increased in sera from Sicilian patients with acute BF compared with healthy controls. sCD8 and sHLA-I reached the peak in the second week after the onset of symptoms, whereas sCD4 and β2-M reached the peak in the first week. Afterwards sCD8 decreased to the levels of controls within the third week, the other parameters decreased later and were unmodified until the third week of infection. Significant correlations were found between sCD4 and sCD8 and the sIL-2R, as well as between serum levels of β2-M and sCD8. The reduction of CD3+ and CD4+ and the increase of CD8+ T cells in the blood indicate that these cells are involved in the response to rickettsia, and their activation might be in part responsible for the release of sCD4 and sCD8. Our data suggest that these soluble markers, indexes of immune activation of T cells both in the circulation and the affected tissues, may be used in monitoring BF evolution.
Expression of Pax5 gene in human haematopoietic cells and tissues: comparison with immunodeficient donors
Authors: Kaneko H , Ariyasu T , Inoue R , Fukao T , Kasahara K , Teramoto T , Matsui E , Hayakawa S , Kondo N
Published: March 2003

In mice, Pax5 gene is indispensable for B cell development. Pax5-deficient mice fail to produce mature B cells owing to complete arrest of B cell development at a precursor stage. However, the lineage and stage of human Pax5 gene expression have remained elusive. In this investigation expression of the human Pax5 gene was studied. Pax5 gene expression was detected in B cell lines but not in myeloma cell lines. CD19 expression was correlated with Pax5 gene expression. Adult spleen and bone marrow and fetal spleen and liver showed strong Pax5 gene expression, as did the corresponding mouse tissues, as reported previously. In common variable immunodeficiency (CVID) peripheral blood lymphocytes (PBL) with a decreased number of B cells, no Pax5 gene expression was detected. Some CVID PBL stimulated with IL-2, IL-10 and anti-CD40 monoclonal antibody, expressed the Pax5 gene. Defect of Pax5 gene expression in CVID may be caused by regulatory T cell disorder.
Cytomegalovirus (CMV) infection in AIDS patients is associated with a CD3 receptor-mediated T cell hyporesponsiveness
Authors: Rowbottom A W , Lepper M W , Sharpstone D , Gazzard B
Published: March 2003

HIV+ individuals with human CMV (HCMV) reactivation have a CD3 receptor-mediated T cell hyporesponsiveness when compared with CD4-matched HIV+ and HCMV−control groups. The impairment of proliferation was not reversed by exogenous IL-2. A typical increase in NFκB expression was observed following cross-linking of the CD3 receptor, but did not lead to increased CD25 cell surface expression or cell proliferation. The HCMV-induced non-responsiveness was not observed when cells were stimulated with phorbol esters. Lymphocytes cultured with media collected from cell cultures infected with HCMV showed a dose-dependent inhibition in the total T cell population even though cells staining dually for CD8/57 increased in number. The altered growth factor requirements of CD8/57+ cells may therefore account for their presence in AIDS and patients following bone marrow transplantation.
Induction of nitric oxide (NO) synthesis in murine macrophages requires potassium channel activity
Authors: Lowry M A R , Goldberg J I , Belosevic M
Published: March 2003

The activation of macrophages for antimicrobial responses is a multistage event involving numerous intracellular signalling cascades that makes possible target cell destruction by these effector cells. This study examined the effects of different potassium channel inhibitors and activators on the NO production of murine macrophage-like cell lines P388D.1 and B10-4(S). We found that the potassium channel inhibitors tetraethylammonium, 4-aminopyridine, and quinine caused dose-dependent reductions in the NO production of macrophages, and that the potassium channel activator, minoxidol, caused a dose-dependent enhancement of NO production. The inhibition of NO production was due to involvement of potassium channels in the priming stage of macrophage activation, since pretreatment with the priming agent interferon-gamma partially restored the NO response of the macrophages. The results of this study demonstrate a link between potassium channel activity and the activation of anitimicrobial functions of murine macrophages.
Effector T lymphocyte subsets in human pancreatic cancer: detection of CD8+ CD18+ cells and CD8+ CD103+ cells by multi-epitope imaging
Authors: Ademmer K , Ebert M , Müller-Ostermeyer F , Friess H , Büchler M W , Schubert W , Malfertheiner P
Published: March 2003

Pancreatic cancer is characterized by an increasing incidence and an extremely poor prognosis. It is resistant to most of the conventional treatment modalities. Histomorphologically, it presents with a strong desmoplastic reaction around cancer cells, and lymphocytes are typically localized as aggregates in the fibrotic interstitial tissue. Using the method of multi-epitope imaging with fluorochrome-tagged specific MoAbs which allows the simultaneous localization and characterization of T cells in tissues, we studied phenotypes and distribution of tumour-infiltrating lymphocytes (TIL) in pancreatic cancer. CD3+ T cells comprised up to 90% of the tumour-infiltrating cells which were either CD4+ or CD8+, most of them being memory cells (CD45RO+). In decreasing order of frequency, T lymphocytes carried the markers for CD45RO, CD18, CD103 and TCR γδ. Very few natural killer cells (CD56+) were observed. Twenty percent of CD8+were labelled with CD103. These CD8+ CD103+T cells, analogous to the gut intraepithelial lymphocytes (IEL), were found in the fibrous interstitial tissue. Furthermore, an inverse correlation was found between the expression of CD18, the β2-integrin, which mediates adhesion of activated lymphocytes, and CD45RO in the CD8+subset of TIL (P = 0.046). In conclusion, phenotyping of T lymphocytes in pancreatic cancer raises the possibility that pancreatic cancer cells develop several strategies to escape the T cell-induced cytolysis by (i) the aggregation of cytotoxic CD8+ CD103+ T cells in the fibrous tissue distant from the tumour cells, and (ii) the presence of CD18-bearing cells which lack the expression of the activation marker CD45RO.
Different susceptibilities of yeasts and conidia ofPenicillium marneffeito nitric oxide (NO)-mediated fungicidal activity of murine macrophages
Authors: Kudeken N , Kawakami K , Saito A
Published: March 2003

Penicillium marneffei is an important opportunistic fungal pathogen. Host defence mechanisms against P. marneffei are not fully understood. We investigated the fungicidal activity of murine peritoneal macrophages against two forms of P. marneffei, conidia and yeast cells, and the involvement of the NO-mediated killing system. Peritoneal macrophages suppressed the intracellular growth of P. marneffei yeast cells and conidia. The number of live yeast cells within macrophages was significantly reduced by activation of macrophages by interferon-gamma (IFN-γ), while a similar response was not observed with conidia. IFN-γ-induced macrophage fungicidal activity against yeast cells was mediated by NO and was almost completely inhibited by NG-monomethyl-l-arginine (l-NMMA), a competitive inhibitor of NO synthesis, while NG-monomethyl-d-arginine (d-NMMA), an optical isomer of l-NMMA, did not show any influence. NO production by macrophages stimulated with IFN-γ was significantly enhanced when these macrophages were cultured with P. marneffei yeast cells, while conidia did not enhance macrophage NO production. Furthermore, yeast cells were more susceptible to the killing effect of chemically generated NO than conidia. Our results indicate that the yeast form of P. marneffei is more sensitive to the fungicidal activity of IFN-γ-stimulated macrophages than conidia, and suggest that the different effects of two forms of P. marneffei on macrophage NO production and their different susceptibilities to NO may be reasons for the present findings.
Prostatein (or rat prostatic steroid binding protein) is a major autoantigen in experimental autoimmune prostatitis
Authors: Maccioni M , Rivero V E , Riera C M
Published: March 2003

Experimental autoimmune prostatitis (EAP) is a disease that could be considered an experimental model of human non-bacterial prostatitis. In this experimental model, male rats are intradermally immunized with a saline extract of male sex accessory glands (RAG) in an adequate adjuvant. The prostatitis observed in the immunized animals develops as a consequence of the immune response against RAG antigens, and the histological lesion is strikingly similar to the pattern of prostatic inflammation observed in the human disease. In this study, we purified one of the prostatic autoantigens recognized by the autoantibodies in our model. Amino acid sequence analysis identified the purified protein as prostatein or rat prostatic steroid binding protein, a member of the uteroglobin superfamily. Prostatein was recognized not only by the humoral autoimmune response, but also by the cellular autoimmune response. Certainly, the DTH response and lymph node cell proliferative assays against prostatein in immunized animals yielded positive results. Prostatein is not only the target of the autoimmune response in animals immunized with the whole extract, but also an inducing antigen of the disease. Purified prostatein, when incorporated to an adequate adjuvant, elicited cellular and humoral autoimmune response and lesion in the prostate gland. The identification of one of the target antigens in autoimmune prostatitis has provided a further refinement and characterization of our model, which could serve for a better understanding of the aetiology, pathogenesis and pathophysiology of non-bacterial prostatitis.
Normal and clonal B lineage cells can be distinguished by their differential expression of B cell antigens and adhesion molecules in peripheral blood from multiple myeloma (MM) patients—diagnostic and clinical implications
Authors: Luque R , Brieva J A , Moreno A , Manzanal A , Escribano L , Villarrubia J , Velasco J L , López-Jiménez J , Cerveró C , Otero M J , Martínez J , Bellas C , Roldán E
Published: March 2003

Human MM is a haematologic disorder characterized by the accumulation of malignant plasma cells (PC), primarily in the bone marrow (BM). Although these cells characteristically home to the BM, in recent years several groups have detected the presence of related malignant B cells in the peripheral blood (PB) which could be implicated in the progression and spread of the disease. However, the proportion and origin of these clonotypic circulating B cells is still controversial. In this study, using a triple-staining flow cytometric procedure and a whole blood lysis method, PB B lineage cells could be divided into two populations according to their distinct repertoires of cell adhesion molecules and B cell antigens in untreated MM patients. The results show that: (i) the percentage and the absolute number of PB CD19+ B cells were decreased in MM patients compared with controls; (ii) the quantity and percentage of B cell antigens (CD20, CD22, CD24, DR, CD138) and adhesion molecules (β1- and β2-integrins, CD44, CD54, CD56, CD61 and CD62L) expressed by these PB CD19+ cells of MM patients and healthy subjects were similar and all of them were virtually polyclonal cells; (iii) a very minor circulating CD19− CD38++ CD45−/dim subset was also detected which expressed CD138 (B-B4) (high intensity), monoclonal cytoplasmic immunoglobulin (cIg), and was negative for pan-B antigens (CD19, CD20, CD24, DR), surface immunoglobulin (sIg) and several adhesion molecules such as CD62L, CD18 and CD11a; this CD19−CD38++ CD45−/dim CD138++ subset was not found in normal blood and exhibited a phenotypic profile which was closely related to that of malignant BM plasma cells, with the exception of the CD56 antigen. Polymerase chain reaction (PCR) analysis of IgH clonotypic rearrangements confirmed these results. We postulate that, in MM patients, circulating B lineage cells may be divided into two different categories: polyclonal CD19+ B cells and a very minor proportion of clonal CD138++ PC that escape from the BM.
Cytokine-enhanced mixed lymphocyte reaction (MLR) in cord blood
Authors: Umemoto M , Azuma E , Hirayama M , Nagai M , Hiratake S , QI J , Kumamoto T , Kobayashi M , Komada Y , Sakurai M
Published: March 2003

Although in cord blood (CB) transplantation graft-versus-host disease (GVHD) is reported to be less severe, GVHD may occur even in patients with HLA-identical sibling donors. This result shows that HLA typing can not entirely predict GVHD. The standard MLR with CB cells was either normal or slightly reduced compared with adult peripheral blood (PB) cells. We used two manipulations to increase the responses of CB cells to allo-antigens. The first was to treat the stimulator cells with cytokines, and the second to amplify weak proliferative responses by adding exogenous cytokines to MLR cultures (modified MLR). The stimulator cells were treated with both interferon-gamma (IFN-γ) and IL-4. The responder cells were treated with both IL-2 and tumour necrosis factor-alpha (TNF-α). It is still to be determined whether or not this cytokine-enhanced MLR could be a possible predictor of GVHD. However, using these cytokines, 90% of CB could recognize allo-antigens, even if the standard MLR was negative.
Circulating antibodies to the 60-kD heat shock protein (hsp) family in patients with Helicobacter pylori infection
Authors: Barton S G R G , Winrow V R , Rampton D S , Crabtree J E , Beales I L P , Calam J
Published: March 2003

Whilst the mechanism by which Helicobacter pylori causes different gastroduodenal diseases is uncertain, strains producing the cytotoxin-associated protein (CagA) have greater pathogenicity. Hsps are immunogenic molecules induced by inflammatory mediators. The aim of this study was to assess pathogenicity of hsp antibodies in H. pylori-infected patients. ELISA techniques were used to assay sera of H. pylori-positive patients with gastritis, gastric atrophy, duodenal or gastric ulcer, and H. pylori-negative controls, for antibodies to CagA and to human, mycobacterial, and in 20 sera, H. pylori (hspB) 60-kD hsp. IgA antibodies to mycobacterial hsp60 in atrophy patients were elevated compared with patients with gastritis (P < 0.05) and with H. pylori-negative controls (P < 0.0005). IgA antibodies to human hsp60 in gastric atrophy patients were elevated compared with H. pylori-negative controls (P < 0.05). Patients with atrophy (P < 0.0005) and gastritis (P < 0.05) who were CagA-positive had raised titres of anti-mycobacterial hsp60 IgA antibodies compared with controls. IgA antibody levels to hspB were positively correlated with those to mycobacterial hsp60 (mhsp60) (P < 0.05) and human hsp60 (hhsp60) (P < 0.005). IgA antibodies to hsp60 are associated with gastroduodenal disease, particularly gastric atrophy, in H. pylori-infected patients. Increased humoral responses to hsp60 could either contribute to gastric atrophy or result from greater gastric mucosal damage induced by CagA-positive strains of H. pylori.