Occupational asthma caused by fish inhalation

Occupational asthma (OA) due to fish inhalation, confirmed by specific bronchial challenge (SBC), has not been described as yet in medical literature, as far as we know. We describe two patients whose asthma was induced by occupational exposure to fish and confirmed by serial measurements of PEFR and SBC. Two fish-processing workers reported asthma symptoms related to their workplace. They were skin tested with fish extracts and their sera assayed for IgE antibodies to various fish species. Nonspecific bronchial reactivity was assessed by methacholine challenge. The occupational relationship was confirmed by PEFR monitoring in working and off-work periods. SBC with fish extracts was carried out to confirm the diagnosis of OA. Skin tests with raw and cooked plaice, salmon, hake, and tuna in patient 1 and anchovy, sardine, trout, salmon, Atlantic pomfret, and sole in patient 2 were positive. Specific IgE serum antibodies were found to salmon in patient 1 and to trout, anchovy, and salmon in patient 2. PEFR measurements differed significantly (P<0.001) between work and off-work periods for both patients. A bronchial challenge with methacholine was positive in patient 1. SBC with raw hake, salmon, plaice, and tuna extracts in patient 1 and raw salmon extract in patient 2 were all positive with an immediate response. SBC with Dermatophagoides pteronyssinus extract was entirely negative in both patients. In three asthmatic, non-fish-allergic controls, SBC with tuna, hake, salmon, and plaice were all negative. These results suggest that fish inhalation can elicit IgE-mediated occupational asthma.     

Exposure to the fish parasite Anisakis causes allergic airway hyperreactivity and dermatitis

BACKGROUND: Several case reports show allergy and anaphylactic reactions to the fish parasite Anisakis in the domestic and occupational setting. Further research is needed on the prevalence and mechanisms of disease. OBJECTIVE: To determine the prevalence of Anisakis sensitization and related symptoms among workers in 2 fish-processing factories, and to use gene-deficient mice to determine the working mechanisms of Anisakis allergy. METHODS: A modified version of the European Community Respiratory Health Survey was used to interview 578 South African fish-processing workers. Sensitization to Anisakis, seafood, and common aeroallergens was determined by skin prick test. Lung function was measured by spirometry and methacholine challenge. Serum eicosapentaenoic acid levels were used as an index of seafood consumption. Sensitized wild-type, IL-4, or IL-4 receptor alpha-deficient mice were challenged orally with Anisakis extract. Allergic reactions, lung pathology, antibodies, cytokines, mast cell proteases, and histamine were evaluated. RESULTS: The prevalence of sensitization to Anisakis was higher than the prevalence of sensitization to fish (8% vs 6%). Anisakis-specific IgE reactivity was associated with bronchial hyperreactivity and dermatitis, and significantly increased with fish consumption. In mice, Anisakis infective larvae (L3) induced a striking T(H)2/type 2 response. Food-allergic-type reactions induced by oral challenge with Anisakis extract were absent in IL-4 receptor alpha knockout mice. CONCLUSION: Anisakis sensitization in fish-processing workers is associated with allergic symptoms and correlates with high levels of fish consumption. Anisakis proteins induce allergic reactions in sensitized mice by IL-4/IL-13-mediated mechanisms. CLINICAL IMPLICATIONS: Anisakis allergy should be considered in fish-processing workers with allergic symptoms.     

Differential responses to natural and recombinant allergens in a murine model of fish allergy

Aerosolized fish proteins are an important cause of allergic airway reactions in both the domestic and the occupational environment. The aim of this study was to investigate inhalant fish-induced allergy in a mouse model and compare immune responses generated by raw and heat-treated fish extracts as well as natural and recombinant forms of the major fish allergen parvalbumin. Mice were sensitized with raw or cooked pilchard extract and challenged intranasally with cooked pilchard extract, purified natural pilchard parvalbumin or recombinant carp parvalbumin (rCyp c1.01). Cooked pilchard extract predominantly sensitized mice to parvalbumin and induced specific IgG1 and IgE antibodies against both pilchard parvalbumin and rCyp c1.01, whereas additional allergens were recognized by mice sensitized with raw extract, including a 36 kDa allergen that was also recognized by fish processing workers and was identified as glyceraldehyde-3-phosphate dehydrogenase. Mice challenged with cooked extract and purified pilchard parvalbumin had increased Th2 cytokine production in mediastinal lymph node cells and splenocytes, whereas mice challenged with rCyp c1.01 did not. This study identifies a new IgE-binding protein that may be important in occupational allergy to fish and demonstrates the feasibility of testing recombinant allergens for immunotherapeutic potential in vivo.     

An immunoassay for verifying the identity of canned sardines

An immunoassay has been developed which facilitates discrimination between canned whole sardine (Sardina pilchardus Walbaum) and other fish species (Pacific pilchard, mackerel, herring, sild and anchovy) that may be substituted for it in the canned product. The non‐competitive indirect ELISA utilizes an antiserum raised against a crude water‐soluble extract of canned sardine. Appropriate antiserum specificity was ensured by using extracts of those fish with which cross‐reactivity was undesirable, to achieve post‐production ‘blocking’ of unwanted cross reactions. This assay will be of value in upholding EC and UK law on the composition and labelling requirements of canned sardine products and provides a basis for the development of a rapid test format.     

Allergy to fish parvalbumins: studies on the cross-reactivity of allergens from 9 commonly consumed fish

BACKGROUND: Fish-hypersensitive patients can probably tolerate some fish species while being allergic to others. OBJECTIVE: To determine the allergenic cross-reactivity between 9 commonly edible fish: cod, salmon, pollack, mackerel, tuna, herring, wolffish, halibut, and flounder. METHODS: Sera from 10 patients allergic to fish and rabbit antisera against 3 parvalbumins (Gad c 1, Sal s 1, and The c 1) were used. Cross-reactivity was investigated by SDS/PAGE and IgE immunoblotting, IgG ELISA, IgE ELISA inhibition, and skin prick test (SPT). RESULTS: Cod (Gad c 1), salmon (Sal s 1), pollack (The c 1), herring, and wolffish share antigenic and allergenic determinants as shown by immunoblots and IgE ELISA, whereas halibut, flounder, tuna, and mackerel displayed lowest cross-reactivities. The highest mean IgE ELISA inhibition percent of 10 sera was obtained by Gad c 1, followed by The c 1, herring, Sal s 1, wolffish, halibut, flounder, tuna, and mackerel with the least inhibition. Nine of the 10 patients showed positive SPT to cod, salmon, and pollack; 8 patients reacted to recombinant (r) Sal s 1. Positive SPTs to rGad c 1 and rThe c 1 were demonstrated in 1 patient. CONCLUSION: Gad c 1, Sal s 1, The c 1, herring, and wolffish contained the most potent cross-reacting allergens, whereas halibut, flounder, tuna, and mackerel were the least allergenic in the current study. The latter could probably be tolerated by some of the tested patients.     

Serologic analysis of dogs, horses, and cottontail rabbits for antibodies to an antigenic flagellar epitope of Borrelia burgdorferi.

Enzyme-linked immunosorbent assays (ELISA) and immunoblots using either whole-cell lysates of Borrelia burgdorferi or an antigenic region of flagellin (41-G) as the antigen were performed, and the abilities of the two assays to detect antibodies to this spirochete in dog, cottontail rabbit, and horse sera were compared. Assays using whole-cell B. burgdorferi lysates as the antigen were more sensitive for detecting antibodies. ELISA with 41-G as the antigen were specific for Borrelia antibodies but were not as sensitive as the assays with whole-cell lysates coated to the solid phase. Use of recombinant full-length flagellin, rather than 41-G, as the antigen in immunoblots increased the sensitivity of each assay. However, antibodies to other bacterial antigens cross-react with whole flagellin and may account for false-positive results. Antibodies to B. burgdorferi outer surface protein A or B were usually undetected when the sera were tested by immunoblotting methods. Borrelia lysates or the 41-G antigen may be used in ELISA or immunoblots to document host exposure to this spirochete. The use of 41-G as the antigen may increase the specificity of an assay or help confirm the serologic diagnosis of Lyme borreliosis in dogs, horses, and cottontail rabbits.     

Characterisation of purified parvalbumin from five fish species and nucleotide sequencing of this major allergen from Pacific pilchard, Sardinops sagax

IgE-mediated allergic reaction to seafood is a common cause of food allergy including anaphylactic reactions. Parvalbumin, the major fish allergen, has been shown to display IgE cross-reactivity among fish species consumed predominantly in Europe and the Far East. However, cross-reactivity studies of parvalbumin from fish species widely consumed in the Southern hemisphere are limited as is data relating to immunological and molecular characterisation. In this study, antigenic cross-reactivity and the presence of oligomers and isomers of parvalbumin from five highly consumed fish species in Southern Africa were assessed by immunoblotting using purified parvalbumin and crude fish extracts. Pilchard (Sardinops sagax) parvalbumin was found to display the strongest IgE reactivity among 10 fish-allergic consumers. The cDNA sequence of the β-form of pilchard parvalbumin was determined and designated Sar sa 1.0101 (accession number FM177701 EMBL/GenBank/DDBJ databases). Oligomeric forms of parvalbumin were observed in all fish species using a monoclonal anti-parvalbumin antibody and subject's sera. Isoforms varied between approximately 10–13 kDa. A highly cross-reactive allergenic isoform of parvalbumin was identified and sequenced, providing a successful primary step towards the generation of a recombinant form that could be used for diagnostic and potential therapeutic use in allergic individuals.     

Airborne dust antigen exposure and specific IgG response in the potato processing industry

Background High prevalences of work-related respiratory symptoms in relation to organic dust exposure have been reported in the potato processing industry, but the responsible effect mechanisms are not known. Objective To study the possible role of a type III allergy in aetiology. Methods Specific immunoglobulin G (IgG) and IgG₄ subclass antibodies against occupational airborne antigens were determined in sera from 131 potato processing workers and 36 non-exposed controls. Personal exposure to airborne antigens was measured, and a preliminary biochemical characterization was carried out. Results Specific IgG was detectable in almost all sera, but levels were significantly (P<0.01) higher in potato processing workers compared with controls. Specific IgG₄ was detectable in half of the workers' sera, but in none of the control sera. The antigens involved appeared to be heat-labile potato proteins. Antibody levels increased during the processing campaign in most workers, and this increase was dependent on the level of antigen exposure. Both the difference in IgG titres between the occupationally exposed group and the non-exposed group, and the exposure-related increase in specific IgG titres seemed to be mainly due to specific antibodies of the IgG₄ subclass. Specific antibodies showed a non-significant tendency to lower levels in workers with work-related respiratory symptoms. Conclusion Occupational respiratory exposure in the potato processing industry leads to a strong humoral immune response, most pronounced for lgG₄ subclass antibodies. Type III allergy is, however, unlikely to play a predominant role in the aetiology of respiratory effects.     

Serologic diagnosis of Lyme borreliosis by using enzyme-linked immunosorbent assays with recombinant antigens

Class-specific enzyme-linked immunosorbent assays (ELISAs) with purified recombinant antigens of Borrelia burgdorferi sensu stricto and Western blot analyses with whole cells of this spirochete were used to test human sera to determine which antigens were diagnostically important. In analyses for immunoglobulin M (IgM) antibodies, 14 (82%) of 17 serum samples from persons who had erythema migrans reacted positively by an ELISA with one or more recombinant antigens. There was frequent antibody reactivity to protein 41-G (p41-G), outer surface protein C (OspC), and OspF antigens. In an ELISA for IgG antibodies, 13 (87%) of 15 serum samples had antibodies to recombinant antigens; reactivity to p22, p39, p41-G, OspC, and OspF antigens was frequent. By both ELISAs, serum specimens positive for OspB, OspE, and p37 were uncommon. Analyses of sera obtained from persons who were suspected of having human granulocytic ehrlichiosis (HGE) but who lacked antibodies to ehrlichiae revealed IgM antibodies to all recombinant antigens of B. burgdorferi except OspB and IgG antibodies to all antigens except OspE. Immunoblotting of sera from the study group of individuals suspected of having HGE reaffirmed antibody reactivity to multiple antigens of B. burgdorferi. There was minor cross-reactivity when sera from healthy subjects or persons who had syphilis, oral infections, or rheumatoid arthritis were tested by ELISAs with p37, p41-G, OspB, OspC, OspE, and OspF antigens. Although the results of class-specific ELISAs with recombinant antigens were comparable to those recorded for assays with whole-cell antigen and for individuals with confirmed clinical diagnoses of Lyme borreliosis, immunoblotting is still advised as an adjunct procedure, particularly when there are low antibody titers by an ELISA.     

Antigenicity and immunocrossreactivity of orange tree pollen and orange fruit allergenic extracts

BACKGROUND: It is important to study the crossreactivity between orange tree pollen (OTPE) and orange fruit (OFE) due to the high incidence of pollen/food-related allergies worldwide. The aim of the present study was to determine the antigenic relationship between OTPE and OFE. METHODS: OTPE and OFErabbit antisera as well as sera from patients allergic to OTPE and OFE were comparatively applied in IgG- and/or IgE-specific ELISA inhibition, crossover or inhibition immunoblotting assays using OTPE and OFE allergenic extracts as solid phase. Periodate and proteinase K treatments were used for carbohydrate and protein depletion, respectively. RESULTS: The antigenicity of OTPE and the presence of common structures between OTPE and OFE extracts were demonstrated by rabbit IgG-specific ELISA inhibition and crossover immunoblotting assays. A 30-kDa protein, common target of the IgE response on OTPE, OFE and mandarin extract, but absent in lemon extract, was identified by ELISA and immunoblot inhibition assays in patients suffering from primary sensitization to OTPE in the context of occupational exposure. Moreover, biochemical treatments showed that antigenic epitopes on the 30-kDa protein contain polypeptidic but no carbohydrate moieties. CONCLUSIONS: The antigenicity of OTPE, the presence of common antigenic determinants between pollen and citrus fruits and an IgE-specific crossreactive protein band of 30 kDa sharing carbohydrate-free epitopes were described. After isolation and purification, this common antigen might be useful for allergen immunotherapy in pollen/fruit-related allergic patients.     

Wheat antigen exposure assessment for epidemiological studies in bakeries using personal dust sampling and inhibition ELISA

Background Asthma in bakery workers caused by exposure to wheat flour proteins is an important occupational health problem. Until recently, gravimetric dust measurements were the only available technique for quantitative exposure assessment in bakeries. However, it is questionable whether dust levels are a good exposure parameter or only give a crude approximation of the actual flour allergen concentration. Objective In the present study we have investigated a method to measure wheat flour antigens with immunochemical methods. Methods Wheat flour antigens were measured in 449 personal dust samples taken in bakeries, using enzyme-linked immunosorbent assay (ELISA) inhibition and an anti-wheat lgG4 serum pool. Western-blotting was performed to compare the wheat flour proteins detected by IgE and IgG4. Results Electrophoresis and immunoblotting showed that many wheat flour proteins can bind lgG4 and IgE, but also a reasonable similarity in major allergens detected by our lgG4-serum pool and IgE-positive sera. Inhibition tests showed some cross-reactivity with some cereal species, but not with other ingredients used in bakeries. In bakeries, large differences in personal airborne flour levels were found between occupational titles. Eor several groups clear differences in wheat antigen exposure levels existed, where no difierences in dust exposure levels could be found. The relationship between dust and wheat antigen exposure varied considerably, depending on the specific bakery occupation, the size of the bakery, and the type of product produced by the bakery. This study also shows that personal sampling of wheat antigens is possible on a large scale and can be used for epidemiological field studies. Conclusion Measurement of airborne wheat antigens in bakeries is a more specific and sensitive measurement tool than measuring dust samples, and will probably be essential for epidemiologic field studies focusing on exposure-response relationships.     

Respiratory and immunological reactions among Shiitake (Lentinus edodes) mushroom workers

Four workers, the total work force employed at a Shiitake farm, developed cough and sputum production following a variable period of exposure to Shiitake mushrooms. All four had abnormal diffusing capacity and three had abnormal spirometry values. Chest roentgenograms demonstrated an interstitial pattern in one worker. Pulmonary function tests performed before and during several days of work demonstrated a significant decrease (greater than 20%) in forced vital capacity (FVC) and/or maximal mid-expiratory flow (MMEF) in three workers. Although specific antibodies to an extract of Shiitake spores were detected in sera from three workers none were IgE. High levels of Shiitake spores were detected in growing rooms (greater than 10(6)/m3) as well as other locations at the farm. Shiitake spore airborne antigen, detected by an immunochemical assay, was present in dust collected with a volumetric sampler from different locations at the farm. Antigenic determinants of Shiitake spore antigens, in common with antigens from other cultivated mushrooms (Agaricus and Pleurotus) were demonstrated by ELISA inhibition assay. This study demonstrates that workers exposed to high levels of Shiitake spores develop symptoms and laboratory findings suggestive of hypersensitivity pneumonitis (HP). Strict environmental control and the wearing of a face mask is probably needed to reduce the high risk of sensitization and possible development of immunological lung disease. Shiitake spores must be considered as an aetiological agent of mushroom workers' lung.     

Allergenic components of Indian jujube (Zizyphus mauritiana) show IgE cross-reactivity with latex allergen

BACKGROUND: 'Latex-fruit syndrome' has been well documented. A prevalence of latex allergy among medical workers of 6.8-8.6% had been reported in Taiwan. However, there has been no study to determine the importance and type of fruit hypersensitivity in latex-allergic patients in Taiwan. This study aimed to identify the allergenic components of Indian jujube (Zizyphus mauritiana) and characterize the cross-reactivity of specific IgE antibodies to latex allergen. METHODS: Crude extracts were prepared from Indian jujube and from ammoniated natural rubber latex, and six medical workers and one patient with a history of fruit allergy underwent skin testing with routine allergens, latex, Indian jujube and other fruits. Sera from two Indian jujube skin test-positive latex-allergic subjects were used for allergen-specific IgE, immunoblotting, immunoblot inhibition and enzyme-linked immunosorbent assay (ELISA) inhibition studies. RESULTS: Both patients had positive skin test responses and specific IgE assays to Indian jujube and latex extracts. Immunoblotting revealed that IgE from both subjects bound to a 42-kD latex protein and a 42-kD Indian jujube protein. In addition, IgE from one subject bound to a 30-kD Indian jujube protein. Preincubation of atopic sera with Indian jujube or latex extract demonstrated absent and/or marked inhibition of IgE binding. Moreover, anti-Indian jujube protein antibody-based ELISA was able to detect latex extracts. CONCLUSIONS: Our results add to findings regarding the 'latex-fruit syndrome' described in the literature, and further study of the cross-reacting allergens identified in Indian jujube may help to elucidate the mechanisms underlying this syndrome.     

Changes in the allergenicity during different preparations of Pomfret, Hilsa, Bhetki and mackerel fish as illustrated by enzyme-linked immunosorbent assay and immunoblotting

BACKGROUND: Although the identification and characterization of several fish allergens have already been reported, there is almost no data on Indian fish allergens and the effect of thermal processing on their allergenicity. This study aimed at the evaluation of the changes in the level of allergenicity of 4 highly consumed Indian fishes, i.e. pomfret, hilsa, bhetki and mackerel, that occurred after boiling and frying. METHODS: In this study 110 patients with fish hypersensitivity as evidenced by clinical history and symptoms were recruited based on their positive skin prick test results. The raw, boiled and fried muscle extracts of the 4 fishes were prepared, and each extract was tested by ELISA and immunoblotting with patients' sera. RESULTS: ELISA and immunoblotting studies demonstrated that the raw muscle extracts of pomfret, hilsa, bhetki and mackerel were allergenic. While the allergenicity of boiled and fried extracts of pomfret and hilsa was considerably reduced, maximum allergenicity of bhetki was demonstrated in the fried extract. The degree of allergenicity of bhetki was demonstrated in the order fried>boiled>raw while that of mackerel followed the order raw>boiled approximately fried. CONCLUSION: The specific IgE-binding activity and immunoblot profile clearly showed that pomfret and hilsa fish allergens are heat-labile, while allergens of bhetki and mackerel maintained strong reactivity even after thermal treatment.     

Food allergy: Specific binding of IgE antibodies from plant food sensitized individuals to carbohydrate epitopes

The presence of anti‐carbohydrate immunoglobulin E (IgE) antibodies was investigated in a group of 54 patients who were sensitive to at least two pollen species and at least five vegetable foods. Enzyme‐linked immunosorbent assays (ELISAs) and ELISA inhibition assays were performed with immobilized food‐related oligosaccharides (e.g. stachyose from legumes) and extracts of plant gums (e.g. gum arabic), as well as with allergen extracts from apples, celery and other fruits and vegetables. Furthermore, a combined procedure of immunoblotting and periodate treatment was applied to identify the epitope nature of glycans from vegetable food glycoproteins. The results suggested that 12/54 of the sera contained IgE antibodies against carbohydrate structures. The antibodies were also found to be highly cross‐reactive. For six patients, galactose may be an important epitope. The clinical relevance of the phenomenon requires further investigation.     

A cell surface ELISA in the mouse using only poly-L-lysine as cell fixative

An ELISA using plates coated with mouse spleen cells has been developed for analysis of antibodies to cell surface antigens. Such assays have been used extensively with human cells or with tumor cells in various species, but application to normal mouse lymphocytes has been limited. Use of normal spleen cells allows access to the genetic resources offered by recombinant, congenic, and mutant mouse strains, in the preparation of cell-coated ELISA plates, the use of glutaraldehyde was found to be unnecessary and it was eliminated, thereby avoiding the destruction of some cell surface determinants. Poly-L-lysine, which was used to treat plates, was found to provide sufficient adherence and preservation of the cells. Binding of biotinylated monoclonal antibodies to cells could be detected at approximately 10 ng/well. In inhibition assays, unlabeled antibodies could be detected at approximately 10 ng/well. Cell-coated plates are stable once prepared, and can be stored for months before use. The assay described can be used to quantitate levels of antibody to a particular epitope, and can also be adapted for screening of fusions for monoclonal antibodies to cell surface antigens.     

Coffee worker's asthma: A clinical appraisal using the radioallergosorbent test

Eight coffee workers with job-related respiratory symptoms were studied with water-soluble green coffee bean (GCB), castor bean (CaB), and factory dust (FD) antigens. Six workers described occupationally related asthma, rhinitis, conjunctivitis, and urticaria or pruritis and demonstrated positive wheal and flare skin tests with GCB and FD antigens. Serum radioallergosorbent test (RAST) indices ranged from 3 to 15 for GCB and 28 to 60 for CaB specificities. The other 2 coffee workers, who denied allergic symptoms, and 8 atopic and sex-matched control subjects demonstrated negative skin tests and RAST indices <2 with these same antigens. Provocative inhalation challenge (PIC) with GCB in 2 skin test-and RAST-positive subjects resulted in significant immediate asthmatic reactions, while PIC in a skin-and RAST-negative asthmatic subject failed to produce an airway response. GCB and CaB antigen characteristics and industrial sources were studied by RAST inhibition analysis. Lack of crossed RAST inhibition with GCB and CaB preparations showed these antigens to be distinct. Several industrial dust and sack samples produced significant RAST inhibition for GCB or CaB determinants. Chlorogenic acid produced no RAST inhibition for either determinant. The results indicated that coffee workers with occupational allergic disease demonstrate serum IgE antibodies specific for etiologic GCB and CaB antigens and that these antigens are distinct, unrelated to chlorogenic acid, present in certain industrial dust and sack samples, and capable of producing asthma in sensitized subjects.     

IgE antibodies of fish allergic patients cross-react with frog parvalbumin

BACKGROUND: The major allergens in fish are parvalbumins. Important immunoglobulin (Ig)E cross-recognition of parvalbumins from different fish species has been shown. Recently frog parvalbumin alpha has been found to be responsible for a case of IgE-mediated anaphylaxis triggered by the ingestion of frog meat. The aim of this study was to investigate whether IgE antibodies of fish allergic persons cross-react with frog parvalbumin and to appreciate its clinical relevance. METHODS: The sera of 15 fish allergic patients and one fish and frog allergic patient were tested by IgE-immunoblotting against frog muscle extract. Sera were tested against recombinant parvalbumin alpha and beta from Rana esculenta. Skin prick tests were performed in selected patients with recombinant frog parvalbumin. Ca(2+) depletion experiments and inhibition studies with purified cod and frog recombinant parvalbumin were done to characterize the cross-reactive pattern. RESULTS: Fourteen of the sera tested had IgE antibodies recognizing low molecular weight components in frog muscle extract. Calcium depletion experiments or inhibition of patient sera with purified cod parvalbumin led to a significant or complete decrease in IgE binding. When tested against recombinant parvalbumins, three of 13 sera reacted with alpha parvalbumin and 11 of 12 reacted with beta parvalbumin from R. esculenta. Skin prick tests performed with recombinant frog parvalbumin were positive in fish allergic patients. Inhibition studies showed that a fish and frog allergic patient was primarily sensitized to fish parvalbumin. CONCLUSION: Cod parvalbumin, a major cross-reactive allergen among different fish species, shares IgE binding epitopes with frog parvalbumin. This in vitro cross-reactivity seems to be also clinically relevant. Parvalbumins probably represent a new family of cross-reactive allergens.     

Human herpesvirus 8 seroconversion in Kenyan women by enzyme-linked immunosorbent assay and immunofluorescence assay.

BACKGROUND: Human herpesvirus 8 (HHV-8) antibody tests vary in reported sensitivity and specificity, depending on the population tested and the assay. OBJECTIVE: The purpose of this study was to compare the ability to detect seroconversion to HHV-8 in a cohort of HHV-8 seronegative female commercial sex workers in Kenya using three tests: HHV-8 viral lysate-based enzyme-linked immunosorbent assay (ELISA), an immunofluorescence assay for HHV-8 lytic antigens (IFA-lytic) and IFA for latent nuclear antigens (IFA-LANA). STUDY DESIGN: By ELISA, 16 women from a prospective cohort of commercial sex workers were identified as seroconverting to HHV-8. A total of 124 post-enrollment samples from these 16 women as well as the enrollment samples were tested for HHV-8 antibodies by all three assays to monitor seroconversion. RESULTS: Of 16 women with apparent seroconversion by ELISA, 8 had a rise in IFA-lytic titers either concomitant with or prior to the first positive ELISA sample and no initial LANA by IFA. Five of the 16 women were IFA-LANA positive at entry, indicating prior infection with HHV-8. Three women had no evidence of seroconversion by either IFA-lytic or IFA-LANA and two of these three had increased ELISA reactivity concomitant with HIV-1 infection. CONCLUSIONS: Conversion from a negative to a positive ELISA result for HHV-8 antibody indicated seroconversion in only half of the study cohort of 16 women when IFA-lytic and IFA-LANA results were considered. The IFA-lytic assay was more sensitive than ELISA for early antibody responses. The IFA-LANA was positive in some women who had neither IFA-lytic nor ELISA antibodies suggesting it may be a marker for latent infections. Presumptive identification of incident HHV-8 infection by ELISA screening followed by IFA-lytic testing to confirm the positive test and IFA-LANA to rule out prior infection provides the most accurate documentation of HHV-8 seroconversion.     

The expert diagnosis of HIV infection by radioimmunoprecipitation]

Using a panel of sera from HIV-infected persons and donors, the authors showed that radioimmunoprecipitation assays compare favourably with immunoblotting assays. With radioimmunoprecipitation, cross reactions were observed between HIV-2 antigens and HIV-2 antibodies, and that the nature of cross reactivity differs from that observed with immunoblotting. Potentials of radioimmunoprecipitation assays as a confirmatory test for use with sera that have given indeterminate results in immunoblotting assays and contradictory results in enzyme immunoassays are examined.