A组乙型链球菌脂磷壁酸的提取及其交叉反应性研究

目的 探讨A组乙型链球菌(iAS)脂磷壁酸(LTA)提取方法及其在风湿热发病过程中的意义。方法 用曲通114(TX—114)从GAS中提取LTA并用阴离子交换树脂二乙基氨基纤维素(DEAE—Sephace1)层析纯化；做LTA与人心瓣膜抗原吸附试验；分析LTA抗体的滴度与风湿性心脏病瓣膜病理结果的关系。结果 用TX—114提取并用阴离子交换树脂DEAE-Sephaccl层析纯化的LTA纯度较高，且保持了LTA的生物学活性；吸附抗体后的血清标本LTA抗体由阳性转为阴性，而末吸附抗体的血清标本LTA抗体仍为阳性，提示LTA与人心瓣膜之间存在交叉抗原性：血清LTA—IgG抗体阳性的4例患者中有3例病理切片发现心瓣膜或心肌间质有灶性炎症细胞浸润；血清LTA-IgG抗体阴性的6例病人均未见灶性炎症细胞浸润(Fiher精确概率=0．033)。结论 TX—114法是提取GAS的LTA的有效新方法；GAS的UFA与人心瓣膜之间存在交叉抗原性，LTA可能参与了风湿热的发病过程。     

Immunological IgE Cross-Reactions of Bovine and Human α-Lactalbumins in Cow's Milk Allergic Patients

Despite great homology with the equivalent human protein, bovine α-lactalbumin (B α-La), a major component of whey, has been identified as a major milk allergen. The aim of this study was to investigate the relationship between structure and IgE binding capacity in α-Las: (1) the importance of three-dimensional structure using native vs disulfide bridgereduced B α-La; and (2) the incidence of amino acid sequence divergence on specific IgE cross-reactivity to human vs bovine α-La. Purified native, reduced and S-carboxymethylated B α-La and human α-La (H alpha-LA) were prepared. Specific IgE of 20 sera from patients with clinically recognized cow's milk protein allergy and positive RAST tests to B α-La were measured in original direct and competitive ELISA inhibition tests. All sera containing specific anti-native B α-La IgE also reacted with denatured protein, but the IgE levels were generally lower, showing that three-dimensional structure is an important feature in B α-La allergenicity but that sequential epitopes are also exposed after protein denaturation. Despite lower IgE levels, all sera also gave significant IgE responses to H α-La. Competitive ELISA inhibition confirmed results obtained by direct ELISA. The demonstrated IgE cross-reactivity between B α-LA and H α-La could be related to the high degree of sequence homology between the two proteins but did not prove to have a clinical significance. However, it is of great interest for a study of the relationship between structure, IgE binding capacity and allergenicity in alpha-Las.     

Allogeneic recognition of class I molecules: Anti-H-2Ld repertoire of H-2b mice includes T cells recognizing mutant class II H-2b (Abm12) molecules

Two major histocompatibility complex (MHC) class I-reactive T cell clones derived from H-2^b mice, generated against the allogeneic L^d molecule, were found to recognize the H-2^b class II mutant A^bm12 molecule as well. In addition, these clones also recognize the class II A^s molecule, and display a class II-dependent reactivity to staphylococcal enterotoxin B. Neither the class I nor the class II alloreactivities of the clones were found to be dependent on other MHC molecules. Both clones express CD4+CD8^? phenotypes. The CD4 molecule appears to be involved in their class II reactivity, while little or no role for CD4 could be detected in the class I reactivity. This is the first report of a class I/class II cross-reactivity being mediated by CD4⁺ T cells. The structural basis for this cross-reactivity is discussed.     

Determination of new cutoff values for indirect immunofluorescence antibody test for Q fever diagnosis in Denmark

Q fever is a ubiquitous zoonosis caused by Coxiella burnetii. The disease is emerging in many parts of the world, likely because of increased awareness and availability of better diagnostics. The diagnosis is primarily based on serology. Because the prevalence of the disease varies worldwide, the establishment of local cutoff values is needed. A baseline for antibodies against C. burnetii in Denmark was defined by testing sera from healthy Danish volunteers using a commercially available immunofluorescence antibody test. Cross-reactivity was studied on sera obtained from patients experiencing clinically related diseases. The cutoff titers suggested by the manufacturer were found to result in very low specificity of the test. The specificity was, however, effectively increased by using cutoff titers based on the local baseline and equal to immunoglobulin M (IgM) phase I ≥128, IgM phase II ≥256, IgG phase I ≥512, and IgG phase II ≥1024.     

Limitations of diagnostic tests for bacterial infections

Lyme disease diagnosis is currently based on serology ? an indirect diagnostic method ? as laboratory cultures are fastidious. The only direct diagnostic method that can be useful with some specimens (cutaneous biopsies or aspiration fluid) is PCR. We aimed to detail the main limitations of serology and PCR testing in the diagnosis of bacterial infections. Limitations are supported by examples from the recent history of microbiology. The main limitation of bacterial serology is the presence of numerous cross-reactions due to many genes that are common to various bacterial species. Some serological techniques, such as those used for the diagnosis of rickettsioses mainly, have even been based on the existence of cross-reactions. The main limitation of PCR testing is the potential presence of laboratory contaminations. PCR-performing laboratories must therefore be certified for the use of this technique. PCR testing also does not inform on the viability of the identified bacterium and should therefore be interpreted in light of the clinical presentation. These limitations highlight that all diagnostic test results should not be interpreted on their own; the clinical and epidemiological contexts should always be taken into consideration.     

Cross-reactivity among evolutionarily distant major histocompatibility complex class I molecules (HLA-B27 and H-2Kk) revealed by xenoreactive T lymphocytes

A set of mouse HLA-B27-reactive cytotoxic T lymphocyte clones were found to recognize the HLA-B27 molecule in an H-2-unrestricted manner, i.e. independently of any mouse major histocompatibility complex (MHC) molecule. The reactivity patterns of these clones on HLA-B27 variants (positive only on HLA-B*2702 and HLA-B*2701) allowed the identification of residues N77 and A81 of the HLA-B27 molecule as important for their reactivity. The location of these residues in the peptide-binding groove (specificity pocket F) suggested that the reactivity of the clones is dependent on HLA-B27-bound peptide(s). However, several other class I molecules sharing these residues (N77 and A81) were not recognized, indicating that other residues might also be involved. One of the clones was found to display an interesting cross-reactivity with allogeneic H-2K^k molecules, sharing N77 and A81 with HLA-B*2702. Sequence comparison suggested the involvement of residue H9, located in specificity pocket B of the peptide-binding groove, and revealed some similarity of pockets B in HLA-B27 and H-2K^k. The structural basis of such T cell-mediated MHC cross-reactions across species barriers is discussed.     

Cross-reactivity between Anisakis simplex sensitization and visceral larva migrans by Toxocara canis

The aim of this work was to study cross-reactivity in the diagnosis of two related ascaridosis. Nineteen patients diagnosed with recidivous acute urticaria (RAU) caused by Anisakis simplex and 26 patients diagnosed with visceral larva migrans (VLM) caused by Toxocara canis were studied employing commercial diagnostic kits and "in house" assay kits. Cross-reactivity observed was greater when using "in house" assay kits, suggesting that T. canis excretory-secretory antigens were not only recognized by antibodies from patients with RAU but with greater intensity compared to the A. simplex excretory-secretory antigens.     

Prevalence of oral allergy syndrome in children with allergic diseases

IntroductionThe oral allergy syndrome (OAS) is a particular type of food allergy rarely explored in the paediatric population that is already considered an adult problem.ObjectiveIdentify the prevalence of OAS, symptoms and pollen species associated with its presence in children affected by allergic diseases.MethodsA cross-sectional study was conducted. Consecutive sampling included children from 6 to 14 years who needed allergy treatment for the first time. A structured questionnaire was carried out to collect demographic and clinical data and history of OAS. Besides sensitisation to various allergens, the skin prick-by-prick test was performed to corroborate sensitisation to food related to OAS. Prevalence of OAS and its association with pollens was established following the covariate adjusted logistic regression.Results267 subjects were included. Overall prevalence of OAS was 8.9% (95%CI 6.1–13.1%). Prevalence of OAS for allergic rhinitis and asthma were 8.8% and 9.1%, respectively. In patients sensitised to pollen, the prevalence ranged from 9.6% to 12.2% depending on the type of pollen. 62.5% of children with OAS were sensitive to pineapple. After adjusting for gender and family history of atopic disease, trees from the Quercus species showed an association with OAS (OR = 2.7, 95%CI 1.2–6.2).ConclusionsOAS is not uncommon in our environment. Pineapple, a typical fruit from the region, was the main food related. Quercus sp., but not birch nor olive, was the pollen associated with this syndrome.     

Clinical and in vitro cross-reactivity of cereal grains in children with IgE-mediated wheat allergy

Introduction and ObjectivesWheat and cereal grains have a broad range of cross-reactivity, but the clinical relevance of this cross-reactivity is uncertain. This study aimed to evaluate clinical and in vitro cross-reactivity with barley, oat, and Job’s tears among wheat-allergic patients.Materials and MethodsPatients aged 5 to 15 years with IgE-mediated wheat allergy were enrolled. Skin prick test (SPT) and specific IgE (sIgE) to wheat, barley, and oat, and SPT to Job’s tears were performed. Oral food challenge (OFC) was conducted if the SPT was ≤5 mm in size and there was no history of anaphylaxis to each grain. Profiles of sIgE bound allergens of wheat, barley, and oat, and inhibition ELISA of IgE binding to barley and oat with wheat were performed.ResultsTen patients with a median age of 8 years were enrolled. Nine of those patients had a history of wheat anaphylaxis. The median SPT size and sIgE level to wheat was 7.3 mm and 146.5 kU_A/l, respectively. The cross-reactivity rate for barley, oat, and Job’s tears was 60.0%, 33.3%, and 20.0%, respectively. Significantly larger SPT size and higher sIgE level were observed in patients with positive cross-reactivity to barley and oat when compared to patients without cross-reactivity. Barley and oat extracts inhibited 59% and 16% of sIgE bound to wheat gliadins and glutenins, respectively.ConclusionThe cross-reactivity rate was quite low for oat and Job’s tears compared to that of barley; therefore, avoidance of all cereal grains may be unnecessary in patients with severe wheat allergy.     

The sensitizing capacity of Compositae plants

Summary Experimental studies in guinea pigs using ether extracts of 20 different species of the Compositae plant family were carried out with the open epicutaneous method (OET) and the guinea pig maximization test (GPMT). The results demonstrate that Cnicus benedictus (blessed thistle), Chrysanthemum leucanthemum (marguerite, ox-eye daisy) and Helianthus debilis (dwarf sunflower) are strong sensitizers while Helenium amarum (bitterweed), Gaillardia amblyodon (blanket flower), Artemisia ludoviciana (prairie sage), Ambrosia trifida (giant ragweed) and Solidago virgaurea (goldenrod) are medium sensitizers. Twelfe species revealed only a weak or no sensitizing capacity; among those were corn flower, wormwood, mugwort, coltsfoot and dandelion. Cross-reactivities were observed in a considerable number of the investigated plant species. The sensitizing power as well as the observed cross-reactions depend on the occurrence of sesquiterpene lactones which have an -methylene group exocyclic to the lactone in common (immunologic requisite). As a practical consequence, patients suffering from allergic contact dermatitis due to Compositae species are strictly requested to avoid contact with the offending species and all related species to prevent recurrences of their skin lesions.