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1.
Transfus Clin Biol ; 29(2): 153-160, 2022 May.
Article in English | MEDLINE | ID: mdl-34856399

ABSTRACT

OBJECTIVES: We aimed to determine a threshold cutoff for hepcidin, ferritin, and the hepcidin-to-ferritin ratio in the diagnosis of liver fibrosis caused by iron overload in chronic hepatitis C virus (HCV)-free ß-thalassemia major patients . METHODS: This 1:1-matched case-control study included 102 individuals (3-30 yr.); 51 ß-thalassemia major patients with iron overload , and 51 apparently healthy individuals. RESULTS: The highest areas under the receiver operating characteristic curves (AUC-ROCs) for the diagnosis of patients vs. controls had overlapping 95% confidence intervals (CIs): serum hepcidin (0.758; 0.64-0.87; P Ë‚ 0.001), serum ferritin (1.000; 1.00-1.00; P˂0.001), and the hepcidin/ferritin ratio (1.000; 1.00-1.00; P˂0.001). For differentiation of patients with liver fibrosis stages of F0-F1 vs. F2-F4 and F0-F1 vs. F3-F4, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) with P-values˂0.001 were the only statistically significant parameters, while the AUC-ROCs of the hepcidin/ferritin ratio (0.631, P=0.188 and 0.684, P=0.098) exhibited 90% and 89.5% sensitivity, respectively, in staging liver fibrosis. CONCLUSION: Our results showed that the hepcidin/ferritin ratio is as effective as the APRI and maybe a better predictor for the diagnosis of liver fibrosis and discriminating its stages, with excellent sensitivity and specificity compared to its components.


Subject(s)
Hepatitis C, Chronic , Iron Overload , beta-Thalassemia , Biomarkers , Case-Control Studies , Ferritins , Hepatitis C, Chronic/complications , Hepatitis C, Chronic/diagnosis , Hepcidins , Humans , Iron Overload/diagnosis , Iron Overload/etiology , Liver Cirrhosis/diagnosis , Liver Cirrhosis/etiology , beta-Thalassemia/complications , beta-Thalassemia/diagnosis
3.
Saudi Med J ; 23(11): 1386-9, 2002 Nov.
Article in English | MEDLINE | ID: mdl-12506301

ABSTRACT

OBJECTIVES: Considering the hazards of exposure to cement dust, this study incorporated basic hematological parameters, erythrocyte sedimentation rate and the total leukocyte count. The idea was to identify a simple, readily available and cost effective screening test that could help in identifying the presence of disease, its severity, or both in cement mill workers potentially related to their work place. METHODS: This study was carried out in the Department of Physiology, Faculty of Health and Medical Sciences, Hamdard University, Karachi and the University of Karachi, Pakistan, during the time frame 1999-2000. In this study a group of 50, apparently healthy volunteers male cement mill workers were randomly selected with ages ranging from 20-60 years. They were matched with another group of 50, control healthy male subjects in terms of age, height, weight and socioeconomic status. Both groups met with exclusion criteria as per standard. The total leukocyte count was performed on an auto analyzer, (Symex-K-1000 CP-Analyzer, Japan) and the erythrocyte sedimentation rate was measured by Westergren tube method. Results were compared in a mean, and on the basis of, period of exposure in a cement mill. RESULTS: In the present study, the mean values of erythrocyte sedimentation rate (p<0.05) and total leukocyte count (p<0.02) significantly increased, but the parameters do not revealing statistically significant differences between 2 groups on the basis of duration of exposure in a cement mill. CONCLUSION: This study has shown that exposure to cement dust causes increased mean values of total leukocyte count and erythrocyte sedimentation rate. However, this change is not related to the duration of exposure in a cement mill.


Subject(s)
Blood Sedimentation , Construction Materials , Leukocyte Count , Occupational Exposure , Adult , Dust , Humans , Male , Middle Aged , Time Factors
4.
J Vet Diagn Invest ; 13(5): 389-93, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11580059

ABSTRACT

A blocking enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies to equine herpesvirus 2 in serum samples of horses. By measuring the binding to a single epitope, this blocking ELISA gives a good picture of the antibody status in the animal. The test is based on a monoclonal antibody with neutralizing activity and had a sensitivity of 94% and a specificity of 100%. Antibodies due to newly acquired infection in foals were successfully detected with this blocking ELISA.


Subject(s)
Antibodies, Viral/analysis , Enzyme-Linked Immunosorbent Assay/veterinary , Herpesviridae Infections/veterinary , Rhadinovirus/immunology , Tumor Virus Infections/veterinary , Animals , Antibodies, Monoclonal , Cell Culture Techniques , Enzyme-Linked Immunosorbent Assay/methods , Female , Herpesviridae Infections/diagnosis , Herpesviridae Infections/immunology , Horses , Male , Rabbits , Sensitivity and Specificity , Tumor Virus Infections/diagnosis , Tumor Virus Infections/immunology
5.
Am J Trop Med Hyg ; 62(1): 106-11, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10761733

ABSTRACT

Puumala (PUU) virus is the causative agent of nephropathia epidemica, the Scandinavian form of hemorrhagic fever with renal syndrome. The infection is acquired by airborne transmission of PUU virus from its rodent reservoir, the bank vole. Besides serologic data indicating that the virus may spread also to heterologous rodents, there is little information on the susceptibility of wild living animals to PUU virus. We studied the occurrence of antibodies to PUU virus in serum samples from 427 wild-living moose, of which 260 originated from the PUU virus-endemic northern and central parts of Sweden and 167 originated from the southern, nonendemic part of Sweden. Samples from 5 animals showed reactivity in an ELISA for recombinant PUU virus nucleocapsid protein, an immunofluorescent assay, and a neutralization test. These 5 animals all originated from the PUU virus-endemic northern part of Sweden. In conclusion, 5 of 260 moose from the endemic region showed convincing serologic evidence of past PUU virus infection. The seroprevalence was low, suggesting that the moose is subjected to endstage infection rather than being part of an enzootic transmission cycle.


Subject(s)
Antibodies, Viral/blood , Deer/virology , Hantavirus Infections/veterinary , Orthohantavirus/immunology , Age Distribution , Animals , Antibodies, Monoclonal , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Fluorescent Antibody Technique, Indirect/veterinary , Hantavirus Infections/epidemiology , Immunoblotting/veterinary , Male , Mice , Mice, Inbred BALB C , Neutralization Tests/veterinary , Nucleocapsid Proteins/immunology , Recombinant Proteins/immunology , Seroepidemiologic Studies , Sweden/epidemiology
6.
Acta Vet Scand ; 40(3): 253-62, 1999.
Article in English | MEDLINE | ID: mdl-10605142

ABSTRACT

An investigation based on 2 studies was carried out to assess the involvement of bovine virus diarrhoea virus (BVDV), bovine herpesvirus type 1 (BHV-1), and bovine respiratory syncytial virus (BRSV) in calf respiratory disease in dairy farms in Venezuela. In the first study, 8 farms were selected and paired serum samples from 42 calves with respiratory disease were tested by ELISA for antibodies to the 3 viruses. Seroconversion to BVDV, BHV-1, and BRSV was found to 5, 2, and 6 farms out of the 8, respectively. The proportion of calves that showed seroconversion to BVDV, BHV-1, and BRSV were 19%, 14%, and 26%, respectively. In the second study, another farm having previous serological evidence of BVDV infection was selected. The decline of maternal antibodies against BVDV was monitored in 20 calves and the half-life of maternal antibodies was 34 +/- 12 days presumably indicating an early natural infection with BVDV. Furthermore, sera free of BVDV antibodies that were collected in studies 1 and 2 and were assayed for the presence of BVDV by nested RT-PCR. Two BVDV strains were detected and compared to those of ruminant and porcine pestiviruses. Both strains were assigned to subgroup Ib of type I BVDV. This investigation provides information on BVDV genotypes circulating in Venezuela and may contribute to the establishment of official control programmes against the viruses studied.


Subject(s)
Bovine Virus Diarrhea-Mucosal Disease/epidemiology , Cattle Diseases/epidemiology , Herpesvirus 1, Bovine/isolation & purification , Respiratory Syncytial Virus Infections/veterinary , Respiratory Syncytial Virus, Bovine/isolation & purification , Spumavirus/isolation & purification , Animals , Antibodies, Viral/isolation & purification , Bovine Virus Diarrhea-Mucosal Disease/blood , Bovine Virus Diarrhea-Mucosal Disease/immunology , Cattle , Cattle Diseases/blood , Cattle Diseases/virology , Dairying , Enzyme-Linked Immunosorbent Assay , Genotype , Herpesvirus 1, Bovine/genetics , Herpesvirus 1, Bovine/immunology , Immunity, Maternally-Acquired , Polymerase Chain Reaction , RNA, Viral/isolation & purification , Respiratory Syncytial Virus Infections/epidemiology , Respiratory Syncytial Virus Infections/immunology , Respiratory Syncytial Virus, Bovine/immunology , Seroepidemiologic Studies , Spumavirus/immunology , Venezuela/epidemiology
7.
Vet Immunol Immunopathol ; 71(1): 41-51, 1999 Oct 01.
Article in English | MEDLINE | ID: mdl-10522785

ABSTRACT

The IgM responses in three panels of sera generated by infection and reinfection of calves with bovine respiratory syncytial virus (BRSV) were measured by indirect ELISA (I-ELISA). The effect of depleting serum IgG by pre-treatment with protein G agarose (PGA) was evaluated. Following primary infection a weak IgM response was detected in the untreated sera of 3 out of 4 calves with maternally derived antibody (MDA). Both the magnitude and duration of the specific IgM responses in these calves were increased by pre-treatment with PGA. In addition, the fourth infected calf tested gave a single positive IgM result following PGA treatment. Transient or persistent IgM responses which were abolished by pre-treatment of sera with PGA were detected in 4/8 calves following reinfection. These were considered to be false positive results, consistent with the influence of IgM rheumatoid factor (IgM-RF). One of these calves and two additional calves showed transient increases in IgM which were resistant to PGA treatment. These were considered to represent specific IgM responses to reinfection. The results indicate the ability of PGA treatment to eliminate both false positive and false negative results and emphasise the necessity for controlling the influence of IgM-RF in IgM-specific indirect ELISAs.


Subject(s)
Antibodies, Viral/analysis , Bacterial Proteins , Cattle Diseases/diagnosis , Enzyme-Linked Immunosorbent Assay/veterinary , Immunoglobulin M/analysis , Respiratory Syncytial Virus Infections/veterinary , Respiratory Syncytial Virus, Bovine/immunology , Sepharose , Animals , Cattle , Chromatography, Affinity/veterinary , False Positive Reactions , Female , Immunoglobulin G/analysis , Immunoglobulin G/isolation & purification , Predictive Value of Tests , Recurrence , Respiratory Syncytial Virus Infections/diagnosis , Sensitivity and Specificity , Streptococcus
8.
Prev Vet Med ; 41(4): 271-8, 1999 Aug 23.
Article in English | MEDLINE | ID: mdl-10530426

ABSTRACT

Six hundred and fifteen serum samples obtained from cows in five districts of Apure State, Venezuela, were tested by ELISA for antibodies to bovine virus diarrhoea virus (BVDV). The same samples were also ELISA-tested for antibodies to bovine herpesvirus type 1 (BHV-1) and bovine respiratory syncytial virus (BRSV). Additionally, the haemagglutination-inhibition (HI) test was used for detecting antibodies to parainfluenza virus type 3 (PIV-3). Overall, seroprevalence to BVDV was 36+/-7% (SE); seroprevalence varied by district (19-42%). BHV-1 seroprevalence was 67+/-4%; variation by district was similar to that of BVDV. However, the first 80 serum samples tested by BHV-1 ELISA all had a strong background reaction with the control antigen. Therefore, these sera were adsorbed to a homogenate of non-infected bovine kidney cell line (MDBK) and retested by ELISA. The non-specific reactivity was significantly reduced (p<0.001 by Wilcoxon's signed-rank test). Compared to the virus-neutralisation (VN) test, the adsorbed BHV-1 ELISA showed 94% agreement and gave a kappa value of 0.84, indicating that the adsorption did not interfere with test accuracy. Seroprevalence against BRSV was 85+/-3%, and showed differences across districts. Most of the cows (94+/-2%) were seropositive to PIV-3, and there were no significant differences among districts.


Subject(s)
Bovine Virus Diarrhea-Mucosal Disease/epidemiology , Diarrhea Viruses, Bovine Viral/immunology , Herpesviridae Infections/epidemiology , Herpesvirus 1, Bovine/immunology , Respiratory Syncytial Virus Infections/veterinary , Respiratory Syncytial Virus, Bovine/immunology , Respirovirus Infections/veterinary , Respirovirus/immunology , Animals , Antibodies, Viral/blood , Bovine Virus Diarrhea-Mucosal Disease/prevention & control , Cattle , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Hemagglutination Inhibition Tests/veterinary , Herpesviridae Infections/prevention & control , Neutralization Tests/veterinary , Respiratory Syncytial Virus Infections/epidemiology , Respiratory Syncytial Virus Infections/prevention & control , Respirovirus Infections/epidemiology , Respirovirus Infections/prevention & control , Seroepidemiologic Studies , Venezuela/epidemiology
9.
Vaccine ; 17(23-24): 2969-73, 1999 Aug 06.
Article in English | MEDLINE | ID: mdl-10462231

ABSTRACT

The potency of 27 different inactivated Newcastle disease (ND) vaccines was determined by immunization and challenge tests. Three- and four-week-old SPF chickens were immunized with 1/50 dose of vaccines. The chickens were bled and challenged three weeks later. The protected/challenged ratio was determined. Serum samples were tested by the haemagglutination inhibition (HI) test and by a monoclonal antibody (MAB) blocking enzyme-linked immunosorbent assay (B-ELISA). Ninety-four percent of the vaccinated chickens (353 birds) with a B-ELISA value above 60% were protected. A strong positive correlation (0.934) was found between 'actual protection' (protection against challenge) and 'estimated protection' (protection calculated from the B-ELISA results). Based upon the results obtained, this B-ELISA seems to be suitable for replacing the challenge test in the potency control of inactivated ND vaccines in the future.


Subject(s)
Antibodies, Blocking/pharmacology , Antibodies, Monoclonal/pharmacology , Newcastle Disease/prevention & control , Newcastle disease virus/immunology , Viral Vaccines/immunology , Animals , Antibodies, Viral/blood , Chickens , Enzyme-Linked Immunosorbent Assay/methods , Hemagglutination Inhibition Tests , Hemagglutination, Viral , Vaccination/veterinary , Vaccines, Inactivated/immunology
10.
J Vet Diagn Invest ; 11(4): 319-23, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10424646

ABSTRACT

A blocking enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies to porcine rubulavirus (La Piedad Michoacan Virus [LPMV]) in serum samples from pigs. The test, based on a monoclonal antibody against the LPMV hemagglutinin-neuraminidase glycoprotein, had a sensitivity of 99% and a specificity of 97%. The results of this test were in agreement with those obtained by an indirect ELISA and hemagglutination inhibition, indirect immunofluorescence, and virus neutralization tests. The blocking ELISA is considered the most suitable test for routine screening for antibodies against LPMV.


Subject(s)
Antibodies, Viral/blood , Rubulavirus Infections/veterinary , Rubulavirus/immunology , Swine Diseases/diagnosis , Animals , Antibodies, Monoclonal , Cell Line , Enzyme-Linked Immunosorbent Assay/methods , Enzyme-Linked Immunosorbent Assay/veterinary , Fluorescent Antibody Technique, Indirect , HN Protein/immunology , Kidney , Mice , Mice, Inbred BALB C , Rubulavirus Infections/diagnosis , Rubulavirus Infections/immunology , Sensitivity and Specificity , Swine , Swine Diseases/blood , Swine Diseases/immunology
11.
J Vet Diagn Invest ; 11(4): 324-9, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10424647

ABSTRACT

An indirect immunofluorescent antibody test (IIFAT) was developed to detect bovine herpesvirus 1 (BHV-1)-specific IgM. All sera were treated with protein-G agarose prior to testing to eliminate the possibility of false-positive results due to IgM-isotype rheumatoid factor (IgM-RF). Specific IgM was first detected 8 days after experimental infection of 3 calves free of maternally derived antibody, with peak responses occurring 2-7 days later. Seroconversion was detected in all 3 calves using a single-dilution enzyme-linked immunosorbent assay. Following reinfection at 30 days postinfection, a low-level IgM response was detected in only 1 calf. Seroconversion was detected in 2 calves. There was no evidence of activation of IgM-RF by infection or reinfection with BHV-1. When 87 acute and convalescent serum pairs collected from 21 outbreaks of respiratory disease were tested, specific IgM was detected in 58 animals (66.6%) from 19 (90.5%) outbreaks. Seroconversion was detected in 44 of these animals (50.6%) from 17 outbreaks (81.0%). The correlations between these 2 assays on a calf and outbreak basis were 79.3% and 90.5%, respectively. Specific IgM was detected in 17/20 sera (85.0%) collected from an additional outbreak. No virus was detected by virus isolation or immunofluorescent staining in nasal mucus samples collected at the same time. Detection of specific IgM by IIFAT is a useful technique for the serodiagnosis of BHV-1 infection.


Subject(s)
Antibodies, Viral/blood , Cattle Diseases/diagnosis , Herpesviridae Infections/veterinary , Herpesvirus 1, Bovine , Immunoglobulin M/blood , Acute Disease , Animals , Antibody Specificity , Cattle , Cattle Diseases/blood , Cattle Diseases/immunology , Convalescence , Enzyme-Linked Immunosorbent Assay , False Positive Reactions , Fluorescent Antibody Technique, Indirect/veterinary , Herpesviridae Infections/diagnosis , Herpesviridae Infections/immunology , Reproducibility of Results
12.
Med Microbiol Immunol ; 187(4): 191-8, 1999 May.
Article in English | MEDLINE | ID: mdl-10363675

ABSTRACT

Immunostimulating complexes (ISCOMs) containing envelope proteins of respiratory syncytial virus (RSV) were explored as a mucosal delivery system for the capacity of inducing a common mucosal antibody response. Two intranasal (i.n.) administrations of BALB/c mice with ISCOMs induced potent serum IgG, and strong IgA responses to RSV locally in the lungs and the upper respiratory, and remotely in the genital and the intestinal tracts. Virtually no measurable IgA response was found in these mucosal organs after two subcutaneous (s.c.) immunizations. Virus neutralizing (VN) antibodies were detected in serum and in all of the mucosal organ extracts after both s.c. and i.n. immunizations indicating that the neutralizing epitopes were preserved after both mucosal and parenteral modes of administration. While the mucosal IgA response appears to be of mucosal origin, the IgG antibodies to RSV detected in the mucosal organs were likely of serum origin. However, the mucosal VN antibodies correlated with the IgG rather than the IgA levels. An enhanced IgA response to gp120 in various mucosal organs was recorded after i.n. immunization with gp120 incorporated in RSV ISCOMs, indicating a role of RSV envelope proteins in enhancing and targeting mucosal responses to passenger antigens.


Subject(s)
Antibodies, Viral/biosynthesis , ISCOMs/immunology , Immunity, Mucosal , Respiratory Syncytial Viruses/immunology , Viral Vaccines/immunology , Animals , Female , ISCOMs/administration & dosage , Immunization , Immunoglobulin A/analysis , Immunoglobulin A/blood , Immunoglobulin G/analysis , Immunoglobulin G/blood , Mice , Mice, Inbred BALB C , Mucous Membrane/immunology , Neutralization Tests , Viral Envelope Proteins/immunology , Viral Vaccines/administration & dosage
13.
J Vet Diagn Invest ; 11(2): 127-33, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10098683

ABSTRACT

Isotype- and subclass-specific indirect enzyme-linked immunosorbent assays were developed to detect parainfluenza-3 virus-specific IgG1, IgG2, IgM, and IgA responses. Sera were treated with protein G-agarose prior to testing for specific IgM and IgA to eliminate the possibility of false-positive results due to IgM-rheumatoid factor and to remove interisotypic competition due to specific IgG. IgM and IgA absorbance values were expressed as a percentage of the absorbance values of positive reference sera included on each plate (S/P%), and respective positive/negative threshold values of 15.0% and 28.0% were determined. The mean interval between experimental infection of 3 calves and initial detection of specific IgG1 and IgG2 responses was 8.0 and 9.3 days respectively, rising rapidly to an initial plateau 13.7 and 11.0 days postinfection (dpi). Reinfection of these calves at 30 dpi resulted in further rapid increases, with higher plateau values reached 13.0 (IgG1) and 13.7 (IgG2) days later. The mean interval between infection and the first positive IgM and IgA responses was 6.7 and 12.3 days, respectively. IgM S/P% values peaked at 13.0 dpi, with all 3 calves showing a secondary anamnestic response to reinfection, peaking 4.7 days later. The IgA response to initial infection was weak, with only 2 calves showing an obvious peak response at 15.0 dpi. A strong anamnestic IgA response to reinfection occurred in 2 calves, with a peak response 9.5 days later. Apparent biphasic and triphasic IgM and IgA responses were evident in some calves. Acute and convalescent serum samples from 80 calves involved in 17 outbreaks of respiratory disease were tested for specific IgM and IgA. Positive IgM results were detected in 15 outbreaks, with 71 sera from 44 calves testing positive. Although IgA-positive results were detected in the same 15 outbreaks, only 42 sera from 31 calves were positive. In a previous study, seroconversion was detected in 21 of these calves from 10 outbreaks. Thus the diagnostic potential of the assays was in the order IgM > IgA > seroconversion. The correlations between IgM and IgA, IgM and seroconversion, and IgA and seroconversion results for each calf were 73.8%, 58.8% and 62.5%, respectively.


Subject(s)
Cattle Diseases/diagnosis , Enzyme-Linked Immunosorbent Assay/veterinary , Immunoglobulin A/analysis , Immunoglobulin M/analysis , Respirovirus Infections/diagnosis , Respirovirus/immunology , Animals , Cattle , Cattle Diseases/immunology , Cattle Diseases/virology , Diagnosis, Differential , Enzyme-Linked Immunosorbent Assay/methods , Immunoglobulin G/analysis , Respirovirus Infections/immunology , Sensitivity and Specificity , Serologic Tests/methods
14.
J Vet Diagn Invest ; 10(4): 331-7, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9786520

ABSTRACT

A commercially available indirect enzyme-linked immunosorbent assay for measuring bovine respiratory syncytial virus (BRSV)-specific IgG was adapted to measure virus-specific IgM. Using this assay, the development of rapid IgM responses in experimentally infected calves was observed 7-9 days postinfection, with peak absorbance values ranging from 1.698 to 2.873. When absorbance values were expressed as a percentage of a positive reference serum, a positive/negative threshold of 22% was determined by testing serum samples from 59 healthy 3-5-month-old calves. Acute and convalescent serum samples collected from 151 calves during 38 outbreaks of respiratory disease were tested, and 130 sera were positive. To determine the number of false-positive results due to the presence of IgM rheumatoid factor, a method for depleting serum IgG by pretreatment of sera with a suspension of protein-G-agarose was developed. All sera that initially tested IgM positive were retested following depletion of serum IgG. False-positive IgM reactions were detected in 23 sera (17.7%). Specific IgM responses were confirmed in 107 sera from 84 calves. Evidence of BRSV infection was detected in 34 of 38 outbreaks. In contrast, seroconversion was detected in 69 calves from 24 outbreaks, confirming the diagnostic potential of the IgM assay. Overall correlation between IgM and seroconversion results was 74.2%. Intra- and interassay reproducibility were 12.50% and 17.48%, respectively (mean coefficients of variation).


Subject(s)
Cattle Diseases/immunology , Enzyme-Linked Immunosorbent Assay/veterinary , Immunoglobulin M/analysis , Respiratory Syncytial Virus Infections/veterinary , Respiratory Syncytial Virus, Bovine/immunology , Rheumatoid Factor/analysis , Animals , Cattle , False Positive Reactions , Reproducibility of Results , Respiratory Syncytial Virus Infections/immunology , Serologic Tests
16.
Clin Exp Immunol ; 113(2): 235-43, 1998 Aug.
Article in English | MEDLINE | ID: mdl-9717973

ABSTRACT

ISCOM is an efficient mucosal delivery system for RSV envelope proteins as measured by antibody responses in respiratory tract secretions and in sera of mice following two intranasal (i.n.) administrations. Intranasally administered RSV ISCOMs induced high levels of IgA antibodies both in the upper respiratory tract and in the lungs. In the lungs, a prominent and long-lasting IgA response was recorded, which still persisted 22 weeks after the second i.n. immunization when the experiment ended. Subcutaneous (s.c.) immunization only induced low IgA titres in the upper respiratory tract and no measurable response to RSV was found in the lungs. Differences were also noticed in serum between the i.n. and s.c. modes of immunization. ISCOMs given intranasally induced earlier, higher and longer lasting IgM and IgG1 serum anti-RSV antibody responses than those induced by the s.c. mode of administration. A low serum IgE response was only detectable at 2 weeks after i.n. immunization with ISCOMs and after s.c. immunization with an inactivated virus, but no IgE response was detectable after s.c. injection of ISCOMs. The serum IgA response was more pronounced following s.c. injection of inactivated virus than after i.n. application of ISCOMs, and a clear-cut booster effect was obtained with a second immunization. Virtually no serum IgA response was detected after the s.c. administration of ISCOMs. In conclusion, the high immune responses induced by RSV ISCOMs in the respiratory tract and serum after i.n. administration indicate prominent mucosal delivery and adjuvant properties of the ISCOMs, warranting further studies.


Subject(s)
Drug Delivery Systems , HN Protein , ISCOMs/administration & dosage , Respiratory Syncytial Viruses/immunology , Respiratory System/immunology , Viral Vaccines/administration & dosage , Administration, Intranasal , Animals , Antibodies, Viral/blood , Antigens, Viral/immunology , Female , ISCOMs/immunology , Immunoglobulin A/analysis , Immunoglobulin G/blood , Immunoglobulin M/blood , Injections, Subcutaneous , Lung/immunology , Mice , Mice, Inbred BALB C , Mucous Membrane/immunology , Nasal Mucosa/immunology , Neutralization Tests , Viral Envelope Proteins/immunology , Viral Proteins/immunology , Viral Vaccines/immunology
17.
J Immunoassay ; 19(2-3): 209-22, 1998.
Article in English | MEDLINE | ID: mdl-9682132

ABSTRACT

Virus neutralization (VN) is an important functional test for evaluating RSV vaccines, also encompassing in mucosal secretion of the respiratory tract considering the infection route. In our previous study, an immunoglobin extraction method described by Bergquist et al. was adopted for RSV ELISA, but it was not suitable for virus neutralization test due to the cell toxicity of the 2% saponin solution used for the antibody extraction. In order to overcome this problem, several solvents including distilled water were tested in the present study for the capacity to extract immunogloblins. Antibodies in the extracts were evaluated and compared by ELISA. Distilled water was as efficient as the 2% saponin solution for extraction of total IgA, RSV specific IgA and IgG. More importantly, the organ extracts obtained subsequently could be used for virus neutralization test without causing adverse effect on the cell culture. Therefore, distilled water was finally chosen as the solvent for immunoglobulin extraction from mucosal organs when both ELISA and virus neutralization test are required.


Subject(s)
Antibodies, Viral/isolation & purification , Immunoglobulin A/isolation & purification , Lung/immunology , Nasal Mucosa/immunology , Neutralization Tests/methods , Respiratory Syncytial Viruses/immunology , Water , Animals , Cell Line , Enzyme-Linked Immunosorbent Assay , Female , ISCOMs/administration & dosage , ISCOMs/immunology , Immunoglobulin G/isolation & purification , Mice , Mice, Inbred BALB C , Mucous Membrane/immunology , Saponins/pharmacology , Statistics, Nonparametric
18.
Vet Immunol Immunopathol ; 61(2-4): 239-50, 1998 Feb 27.
Article in English | MEDLINE | ID: mdl-9613438

ABSTRACT

Sera from 19 colostrum-deprived calves less than 1 week old, 24 colostrum-supplemented calves less than 1 week old, 36 3-5-month-old calves and 200 females greater than 9 months of age were tested by ELISA for the presence of IgM, IgG and IgA rheumatoid factors (RF). An increasing level of IgM- and IgG-RF with age was found. IgG-RF levels in the colostrum-supplemented calves were significantly higher than in the non-supplemented calves (p < 0.001). Individual IgG-RF values correlated with serum IgG levels, as determined by zinc sulphate turbidity testing (r=0.59, p < 0.01). No IgA-RF was detected. The cross-reactivity of IgM-RF with heterologous IgG was found to be greatest with rabbit IgG, followed by mouse and chicken IgG. The significance of rheumatoid factors in relation to diagnostic testing is discussed.


Subject(s)
Cattle/immunology , Enzyme-Linked Immunosorbent Assay/veterinary , Rheumatoid Factor/blood , Animals , Chickens , Colostrum/immunology , Cross Reactions , Enzyme-Linked Immunosorbent Assay/methods , Female , Immunity, Maternally-Acquired , Immunoglobulin A/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , Mercaptoethanol , Mice , Nephelometry and Turbidimetry , Rabbits , Species Specificity , Zinc Sulfate
19.
Acta Vet Scand ; 39(1): 25-33, 1998.
Article in English | MEDLINE | ID: mdl-9592943

ABSTRACT

Eighty Standardbred horses, originating from 5 training campuses, with decreased athletic performance in association with symptoms such as intermittent fever and mild pharyngitis were examined. As control animals, 10 horses from a stable with normally performing horses were used. Virus isolation and clinico-chemical and serological tests were performed. Lymphocyte proliferation tests were carried out to evaluate the capacity of the cell-mediated immunity. In addition, a bioassay for equine type I interferon, as a marker for early viral infections, was established. No specific microbe could be linked to these symptoms, but there was a temporary suppression of the cell-mediated immunity, which might be explained by the serological evidence of an EHV-2 and/or rhinovirus infection.


Subject(s)
Horse Diseases , Horses/immunology , Lymphocytes/immunology , Physical Conditioning, Animal , Picornaviridae Infections/veterinary , Picornaviridae , Animals , Antibodies, Viral/blood , Fever/immunology , Fever/veterinary , Immunity, Cellular , Interferon-alpha/biosynthesis , Lymphocyte Activation , Pharyngitis/immunology , Pharyngitis/veterinary , Picornaviridae Infections/blood , Picornaviridae Infections/immunology , Running
20.
J Vet Diagn Invest ; 10(1): 43-8, 1998 Jan.
Article in English | MEDLINE | ID: mdl-9526859

ABSTRACT

A single-dilution quantitative enzyme-linked immunosorbent assay (ELISA) system, based on commercial ELISA kits, for the simultaneous detection of seroconversion to bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), parainfluenza-3 virus (PI3V), and infectious bovine rhinotracheitis virus (IBRV) was evaluated by testing acute and convalescent serum pairs from 564 cattle in 145 outbreaks of respiratory disease. Seroconversion to BVDV, BRSV, PI3V and IBRV was detected in 8.0%, 19.0%, 13.7%, and 7.4%, respectively, of serum pairs tested. Seroconversion was detected in 60.7% of herds and 34.6% of animals tested. Infection with 2 or more viruses was found in 46.6% of these herds and in 27.2% of these animals. The majority of BVDV infections (62%) were associated with other virus infections, suggesting that BVDV may potentiate infection with other agents rather than being a primary pathogen of the respiratory tract. The results were compared with those obtained by virus neutralization and hemagglutination inhibition testing, and the sensitivity, specificity, and overall correlation were calculated. Sensitivities of 92%, 95%, 100%, and 100% were obtained for BVDV, BRSV, PI3V, and IBRV, respectively. The corresponding specificity values were 89%, 92%, 86%, and 91%. The overall correlation for each virus was 90%, 93%, 90%, and 93%, respectively. These results demonstrate that this ELISA system may be used successfully to detect seroconversion in serum pairs, highlight the frequency of multiple viral infections in outbreaks of respiratory disease, and provide further evidence of an immunosuppressive role for BVDV infections.


Subject(s)
Bovine Virus Diarrhea-Mucosal Disease/diagnosis , Cattle Diseases , Infectious Bovine Rhinotracheitis/diagnosis , Respiratory Syncytial Virus Infections/veterinary , Respiratory Syncytial Virus, Bovine , Respirovirus Infections/veterinary , Animals , Bovine Virus Diarrhea-Mucosal Disease/epidemiology , Cattle , Diagnosis, Differential , Disease Outbreaks/veterinary , Enzyme-Linked Immunosorbent Assay/methods , Hemagglutination Inhibition Tests/methods , Infectious Bovine Rhinotracheitis/epidemiology , Neutralization Tests/methods , Respiratory Syncytial Virus Infections/diagnosis , Respiratory Syncytial Virus Infections/epidemiology , Respirovirus Infections/diagnosis , Respirovirus Infections/epidemiology
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