Two monoclonal antibody probes were produced against PK'X' cells. The parasites were partially purified by filtration and centrifugation of kidney tissue from rainbow trout with proliferative kidney disease and used as antigen for immunization of mice. The resulting monoclonal antibodies reaeted with PK'X' eell antigens in enzyme-linked immunosorbent assay and immunohistoehemistry tests. One antibody (Mab 12) appeared to be specific for PK'X' in kidney tissue, while the other {Mab 18) cross-reacted with host eell antigens in the kidney tubules. These probes are invaluable tools for the investigation of parasite surface antigens and life cyele studies.
The lectin GS-I (Griffonia siniplicifol~a agglut~nin-I) and a recently developed monoclonal antibody (Mabl2) both recognized extrasporogonic (interstitial) and sporogonic (intraluminal) stages of the PKX myxosporean causing proliferative kidney disease (PKD) in salmonid fish. In histochemical and immunostains the lectin and antibody recognized PKX from North American and European salmonids including Atlantic salmon Salmo salar, chinook salmon Oncorliynchus tshawytscha, coho salmon Oncorhynchus kisutch, brown trout Salmo trutta, and grayling Thymallus thymallus. Although the specific structures recognized by the lectin and Mabl2 were not determined, double fluorescent staining techniques showed similar staining patterns with extrasporogonic or interstitial stages of the PKX myxosporean. An apparent inhibition between the lectin and Mabl2 in these double stains suggests that they recognize similar structures of the parasite. Sporogonic stages of PKX, some containing polar capsules, were detected with the lectin and Mabl2 stains in fish with typical extrasporogonic (interstitial) P M providing further confirmation of the relatedness of these stages. Both the lectin and 1Mabl2 provide useful tools for further study of the epidemiology of the PKX myxosporean, for future development of tests to detect anti-parasite antibodies in fish serum and for the identification of potential imrnunogens for vaccination of trout.
The results of comparative immunohisto-chemical staining studies with two lectin probes, GS-I and SBA, and five monoclonal antibodies raised against the PKX myxosporean are presented. In order to investigate the postulated link between PKX and Sphaerospora spp., the lectins and monoclonal antibodies were tested against PKX, Sphaerospora renicola, Sphaerospora oncorhynchi, Sphaerospora eiegans, Sphaerospora truttae and Sphaero-spora sp. from Atlantic salmon, Salmo salar L., and Ceratomyxa shasta. SBA appears a useful tool for the diagnosis of all Sphaerospora spp. infections and was also positive for C. shasta. GS-I and monoclonal antibodies (12, C3, D4 and A3) appear to be specific for both the extrasporogonic and sporogonie stages of PKX. Although MabB4 was raised to PKX, it appears to stain some of the sporogonie stages of Sphaerospora spp. but not C shasta. The results of the lectin and monoclonal staining of PKX and Sphaerospora oncorhynchi indicated that further work was required to prove or disprove the hypothesis that PKX and S. oncorhynchi are the same organism.
BackgroundThe specific active immunotherapy, employing vaccine of allergen of mite is a treatment considered as effective for the respiratory allergy and asthma. The sublingual route has minor risk of systematises reactions. The objective of this study was to determine the therapeutic effect and security of sublingual immunotherapy (ITSL) employing the standard vaccine VALERGEN-DP (BIOCEN, CUBA) in a population of asthmatic Cuban patients.MethodsA phase II Clinical Trials double blind, placebo controlled in a total of 40 adult patients with mild or moderate asthma and specific sensibility preponderant to this mite. Half of patients received drops by sublingual route with growing doses up to 2000 UB.ResultsThe treatment was effective in the reduction of clinical symptoms and medication intake as compared to conventional treatment in control group. The cutaneous sensibility to this mite was significant reduced, increasing in 1.9 log; the amount of necessary allergen to provoke a positive Prick Test. An improvement of the lung function was observed with a significant reduction (P < 0.05) of expiratory pick flow variability. The frequency of local reactions were only 0.58% of administration.ConclusionsThe VALERGEN-DP vaccine is an effective treatment and profitable against asthma in our population and guarantee its generalization in the Allergy Services of our health system.
BackgroundAllergen immunotherapy (AIT) induces IgG4 antibodies with blocking effect, and long-term reduction of IgE. The problem of finding suitable surrogate paraclinical markers during AIT is currently very relevant. The aim was to evaluate the allergen-specific IgE/IgG4 ratio as an immunological correlate to clinical efficacy during AIT with House Dust Mite (HDM) allergen vaccines in Cuban asthmatic patients.MethodsThree separate Double-Blind Placebo-Controlled clinical trials of AIT were performed using standardized registered allergen extracts of 3 HDM species: Dermatophagoides pteronyssinus, and the tropical species D. siboney and Blomia tropicalis. Each clinical trial included 40 asthmatic patients, totalling 120 individuals. Half of them received placebo. Allergen-specific IgE and IgG4 antibodies were measured by ELISA. Antibody titres were normalized and averaged between the 3 trials. Size effect was calculated as the Standard Mean Difference (SMD) between the active and placebo groups, using meta-analysis tools.ResultsThere was a significant increase of IgG4 antibodies (P < 0.05) after 6 months of treatment. At 12 months, the IgG4 increase was even greater and the IgE decrease achieved also significance (P < 0.05). Thus, IgG4 induction appeared to precede IgE changes, in agreement with the possible role of the Treg/IL-10 response induction at the initial AIT phase. Moreover, 83% of patients showed a decrease of the IgE/IgG4 ratio in the active groups, whereas only 32% showed reduction in placebo groups. The IgE/IgG4 ratio was the immunological variable with the greatest size effect value (SMD = 0.81 95% CI, 0.71-0.91), as compared to changes in IgE or IgG4 levels, alone. The size effect value was close to the clinical effect (symptom-medication score SMD = 1.2 95% CI, 0.7-1.7). Per-patient changes in IgG4 levels, as well as, in the IgE/IgG4 ratio, were significantly correlated to the symptom-medication variable (r = 0.23, P = 0.04).ConclusionsThese results support the use of the IgE/IgG4 ratio as an easily measurable marker for monitoring the allergen-specific immunity during AIT with HDM in asthmatic patients, and possibly, for predicting patient's clinical improvement after 1 year of treatment.
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