scholarly journals Antibody Responses in Reindeer (Rangifer tarandus) Infected with Mycobacterium bovis

2005 ◽  
Vol 12 (6) ◽  
pp. 727-735 ◽  
Author(s):  
W. R. Waters ◽  
M. V. Palmer ◽  
J. P. Bannantine ◽  
R. Greenwald ◽  
J. Esfandiari ◽  
...  

ABSTRACT Despite having a very low incidence of disease, reindeer (Rangifer tarandus) are subject to tuberculosis (TB) testing requirements for interstate shipment and herd accreditation in the United States. Improved TB tests are desperately needed, as many reindeer are falsely classified as reactors by current testing procedures. Sera collected sequentially from 11 (experimentally) Mycobacterium bovis-infected reindeer and 4 noninfected reindeer were evaluated by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and multiantigen print immunoassay (MAPIA) for antibody specific to M. bovis antigens. Specific antibody was detected as early as 4 weeks after challenge with M. bovis. By MAPIA, sera were tested with 12 native and recombinant antigens, which were used to coat nitrocellulose. All M. bovis-infected reindeer developed responses to MPB83 and a fusion protein, Acr1/MPB83, and 9/11 had responses to MPB70. Other antigens less commonly recognized included MPB59, ESAT-6, and CFP10. Administration of purified protein derivatives for skin testing boosted serum antibody responses, as detected by each of the assays. Of the noninfected reindeer, 2/4 had responses that were detectable immediately following skin testing, which correlated with pathological findings (i.e., presence of granulomatous lesions yet the absence of acid-fast bacteria). The levels of specific antibody produced by infected reindeer appeared to be associated with disease progression but not with cell-mediated immunity. These findings indicate that M. bovis infection of reindeer elicits an antibody response to multiple antigens that can be boosted by skin testing. Serological tests using carefully selected specific antigens have potential for early detection of infections in reindeer.

2013 ◽  
Vol 20 (6) ◽  
pp. 907-911 ◽  
Author(s):  
Konstantin P. Lyashchenko ◽  
Rena Greenwald ◽  
Javan Esfandiari ◽  
Daniel J. O'Brien ◽  
Stephen M. Schmitt ◽  
...  

ABSTRACTBovine tuberculosis (TB) in cervids remains a significant problem affecting farmed herds and wild populations. Traditional skin testing has serious limitations in certain species, whereas emerging serological assays showed promising diagnostic performance. The recently developed immunochromatographic dual-path platform (DPP) VetTB assay has two antigen bands, T1 (MPB83 protein) and T2 (CFP10/ESAT-6 fusion protein), for antibody detection. We evaluated the diagnostic accuracy of this test by using serum samples collected from groups of white-tailed deer experimentally inoculated withMycobacterium bovis,M. aviumsubsp.paratuberculosis, orM. bovisBCG Pasteur. In addition, we used serum samples from farmed white-tailed deer in herds with no history of TB, as well as from free-ranging white-tailed deer culled during field surveillance studies performed in Michigan known to have bovine TB in the wild deer population. The DPP VetTB assay detected antibody responses in 58.1% of experimentally infected animals within 8 to 16 weeks postinoculation and in 71.9% of naturally infected deer, resulting in an estimated test sensitivity of 65.1% and a specificity of 97.8%. The higher seroreactivity found in deer with naturally acquiredM. bovisinfection was associated with an increased frequency of antibody responses to the ESAT-6 and CFP10 proteins, resulting in a greater contribution of these antigens, in addition to MPB83, to the detection of seropositive animals, compared with experimentalM. bovisinfection. Deer experimentally inoculated with eitherM. aviumsubsp.paratuberculosisorM. bovisBCG Pasteur did not produce cross-reactive antibodies that could be detected by the DPP VetTB assay. The present findings demonstrate the relatively high diagnostic accuracy of the DPP VetTB test for white-tailed deer, especially in the detection of naturally infected animals.


2015 ◽  
Vol 22 (6) ◽  
pp. 641-649 ◽  
Author(s):  
W. Ray Waters ◽  
Mitchell V. Palmer ◽  
Molly R. Stafne ◽  
Kristin E. Bass ◽  
Mayara F. Maggioli ◽  
...  

ABSTRACTSeveral serological tests designed to detect antibodies to immunodominantMycobacterium bovisantigens have recently emerged as ancillary tests for the detection of bovine tuberculosis in cattle, particularly when used after the injection of purified protein derivative (PPD) for skin testing, which significantly boostsM. bovis-specific antibody responses. The present findings demonstrate the onset and duration of boosted antibody responses after the injection ofM. bovisPPD for the caudal fold test (CFT) andMycobacterium aviumandM. bovisPPDs for the comparative cervical test (CCT), administered in series in cattle experimentally infected withM. bovis. While skin tests boosted the responses to certain antigens (i.e., MPB83 and MPB70), they did not affect the responses to other antigens (e.g., ESAT-6, CFP10, MPB59, and MPB64). Administration of the CCT 105 days after the CFT resulted in an even greater secondary boost in antibody responses to MPB83 and MPB70 and to a proteinase K-digested whole-cell sonicate (WCS-PK) ofM. bovis. Both IgM and IgG contributed to the initial boost in the MPB83/MPB70-specific antibody response after the CFT. The secondary boost after the CCT was primarily due to increased IgG levels. Also, the avidity of antibodies to MPB83 and MPB70 increased after the CCT inM. bovis-infected cattle. The avidity of antibodies to the WCS-PK antigens increased in the interval between the CFT and the CCT but did not increase further after the CCT. Together, these findings demonstrate that the administration of PPDs for skin tests results in additive enhancement (i.e., when the CFT and CCT are performed in series), both qualitative and quantitative, of MPB83/MPB70-specific antibody responses.


2011 ◽  
Vol 18 (12) ◽  
pp. 2154-2160 ◽  
Author(s):  
Fangui Min ◽  
Yu Zhang ◽  
Ren Huang ◽  
Wende Li ◽  
Yu'e Wu ◽  
...  

ABSTRACTOld tuberculin (OT) and purified protein derivative (PPD) are widely used for tuberculin skin testing (TST) in diagnosis of tuberculosis (TB) but often yield poor specificity and anergy in reaction. Therefore, it is necessary to develop new serological methods as a possible auxiliary diagnostic method for TB. In this study, we characterized the dynamic antibody responses of 10 purified recombinant antigens, PPD, and OT in rhesus monkeys experimentally infected withMycobacterium tuberculosisand analyzed the time to antibody detection, antibody levels, and their association with the infectious doses. The antibodies were detected as early as 4 weeks after infection in response to 5 antigens (CFP10, CFP10-ESAT-6, U1, MPT64, and Ag85b). Antibodies against most of the other antigens were detected between 4 and 12 weeks after infection. The levels of antibodies were dose dependant. We further evaluated the serodiagnostic potential of these antigens by using indirect enzyme-linked immunosorbent assay in 71 TST-positive and 90 TST-negative serum samples from monkeys. For all 12 antigens, the median optical density values of TST-positive monkeys were statistically significantly higher than those of TST-negative monkeys (P< 0.001). Among those antigens, Ag85b and CFP10 showed higher diagnostic potential than others. A combination of results from Ag85b, the 38-kDa antigen (Ag38kDa), and Ag14kDa reaches a sensitivity of 95.77%, indicating that these antigens may be ideal cocktails in TB diagnosis.


2008 ◽  
Vol 16 (2) ◽  
pp. 209-217 ◽  
Author(s):  
David J. Wilson ◽  
Bonnie A. Mallard ◽  
Jeanne L. Burton ◽  
Ynte H. Schukken ◽  
Yrjo T. Grohn

ABSTRACT Dairy cattle in two commercial Holstein herds were randomly selected to be vaccinated twice with J5, at approximately 60 days and 28 days before the expected calving date, or to be untreated controls. Based on whether milk production changed following clinical mastitis or whether cows were culled or died within 30 days after onset, 51 mastitis cases were classified as severe or mild. J5-specific antibody responses were evaluated by enzyme-linked immunosorbent assay of all 32 severe and 19 mild cases. The amounts of J5-specific immunoglobulin M (IgM), IgG1, and IgG2 antibodies in sera from the 27 J5 vaccinates were compared with those of the 24 controls. At drying off (before J5 vaccination), all cows had similar amounts of J5-specific antibody. Immediately after calving (approximately 28 days after the second vaccination), J5 vaccinates had significantly higher production of J5-specific IgG1 and IgG2 than controls. When cows were tested following clinical mastitis, none of the three antibody classes differed significantly between the controls and the vaccinates. Vaccinates that contracted Escherichia coli mastitis had 75% less milk loss than controls. The cows that contracted clinical mastitis later in lactation, the unvaccinated controls, and those infected with E. coli had more milk loss following mastitis. The hazards of being culled for all reasons and of being culled for mastitis were significantly lower for J5 vaccinates. Vaccination with J5 was associated with protection against milk production loss and culling following clinical mastitis, and it was also significantly associated with changes in J5-specific IgM, IgG1, and IgG2 antibodies in sera of vaccinated cows.


2004 ◽  
Vol 72 (5) ◽  
pp. 2462-2467 ◽  
Author(s):  
Konstantin Lyashchenko ◽  
Adam O. Whelan ◽  
Rena Greenwald ◽  
John M. Pollock ◽  
Peter Andersen ◽  
...  

ABSTRACT Vaccine development and our understanding of the pathology of bovine tuberculosis in cattle would be greatly facilitated by definition of the immunological correlates of protection and/or pathology. In this study we analyzed humoral immune responses in Mycobacterium bovis BCG-vaccinated and control cattle (in particular, the relationship between the intradermal comparative tuberculin skin test and serum immunoglobulin G [IgG] responses) against a range of mycobacterial antigens (MPB59, MPB64, MPB70, MPB83, ESAT-6, CFP-10, Acr1, and PstS-1) by multiantigen print immunoassay and conventional enzyme-linked immunosorbent assay. Following M. bovis infection, the comparative tuberculin skin test strongly boosted IgG, IgG1, and IgG2 antibody responses, particularly against MPB83 and MPB70, in unvaccinated cattle but failed to boost these responses, or did so only weakly, in BCG-vaccinated calves. In addition, the skin test-induced increases in MPB83-specific IgG responses correlated positively with bacterial loads and ESAT-6-induced in vitro gamma interferon responses. In conclusion, both the negative correlation of skin test-enhanced MPB83-specific antibody responses with BCG-induced protection and their positive correlation with bacterial loads can serve as useful markers for vaccine efficacy after challenge.


2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S206-S206
Author(s):  
Kasturi Banerjee ◽  
Michael Motley ◽  
Elizabeth Diago-Navarro ◽  
Bettina C Fries

Abstract Background Capsular polysaccharide (CPS) of Carbapenem-resistant K. pneumoniae ST258 (CR-Kp) is a potential vaccine target. CPS of these isolates generally falls within 2 homology groups named clade 1 and clade 2. We and others have made antibodies (Abs) that act against clade2 CR-Kp but failed to make therapeutic Abs against clade1 CR-Kp. Previous studies had shown that studying patient’s antibody responses could help in identifying suitable candidates for developing immunotherapies. Thus, we sought to identify potential vaccine candidates by investigating the humoral response CPS in CR-Kp-infected patients. Methods 24 CR-Kp isolates and corresponding serums were collected from inpatients at Stony Brook Hospital. CPS was isolated and purified by size-exclusion column chromatography from CR-Kp strains 34 (clade 2), 36 (clade 1), and 38 (clade-Other). Anti-CPS Abs in patient’s serum were detected by enzyme-linked immunosorbent assay (ELISA) and bulk Abs from positive serum were purified using an affinity column. These Abs were tested for activity against CR-Kp by serum bactericidal and agglutination assays. Results 50% of clade2 CR-Kp-infected patients had humoral responses against CPS34. 77% of clade 1-infected patients sera cross-reacted wtih CPS34, but none of them developed Abs against CPS36. Interestingly, 90% of clade1 and 60% of clade 2-infected patients, respectively, showed Abs binding to CPS38. Thus, we selectively purified Anti-CPS Abs from two clade-Other-infected patients and observed that they were cross-reactive with all three CPS. Further, these Anti-CPS Abs agglutinated all tested CR-Kp isolates (34, 36, and 38) when compared with control human IgG (P < 0.005). Additionally, these Anti-CPS Abs promoted killing of clade2 bacteria and inhibited the growth of clade1 bacteria in Ab-mediated serum bactericidal assay. These data elucidate that humoral responses developed in clade-Other CR-Kp-infected patients have therapeutic potential. Conclusion With the unavailability of effective antimicrobials for CR-Kp, approaches like developing novel anti-CPS vaccine could serve as an alternate therapy. Our data suggest that developing immunotherapies targeting CPS38 could potentially provide protection across both clade1 and clade2 bacteria in clinical settings. Disclosures All authors: No reported disclosures.


2012 ◽  
Vol 19 (4) ◽  
pp. 527-535 ◽  
Author(s):  
Bettina Wagner ◽  
Heather Freer ◽  
Alicia Rollins ◽  
David Garcia-Tapia ◽  
Hollis N. Erb ◽  
...  

ABSTRACTLyme disease in the United States is caused byBorrelia burgdorferisensu stricto, which is transmitted to mammals by infected ticks.Borreliaspirochetes differentially express immunogenic outer surface proteins (Osp). Our aim was to evaluate antibody responses to Osp antigens to aid the diagnosis of early infection and the management of Lyme disease. We analyzed antibody responses during the first 3 months after the experimental infection of dogs using a novel multiplex assay. Results were compared to those obtained with two commercial assays detecting C6 antigen. Multiplex analysis identified antibodies to OspC and C6 as early as 3 weeks postinfection (p.i.) and those to OspF by 5 weeks p.i. Antibodies to C6 and OspF increased throughout the study, while antibodies to OspC peaked between 7 and 11 weeks p.i. and declined thereafter. A short-term antibody response to OspA was observed in 3/8 experimentally infected dogs on day 21 p.i. Quant C6 enzyme-linked immunosorbent assay (ELISA) results matched multiplex results during the first 7 weeks p.i.; however, antibody levels subsequently declined by up to 29%. Immune responses then were analyzed in sera from 125 client-owned dogs and revealed high agreement between antibodies to OspF and C6 as robust markers for infection. Results from canine patient sera supported that OspC is an early infection marker and antibodies to OspC decline over time. The onset and decline of antibody responses toB. burgdorferiOsp antigens and C6 reflect their differential expression during infection. They provide valuable tools to determine the stage of infection, treatment outcomes, and vaccination status in dogs.


Author(s):  
ZM Huo ◽  
J Miles ◽  
PG Riches ◽  
T Harris

Background: Measurement of antibody responses to polysaccharide antigens is regarded as an important assessment of an individual's ability to respond to carbohydrate antigens. The currently used assays for the measurement of pneumococcal-specific antibody use the multi-serotype vaccine Pneumovax® as the detection antigen. Methods: An equal potency enzyme-linked immunosorbent assay (ELISA) system was used to compare the measurement of serotype-specific antibody with the multi-serotype assay. Results: Our results show that the concentration of specific antibody to Pneumovax is not related to the concentration of antibody to the individual serotypes. Neither is any correlation found between the antibody concentrations to any of the three single serotypes investigated, to the mixture of the three serotypes or to Pneumovax. Conclusion: We conclude that the measurement of the concentration of the specific antibody to the mixed serotypes present in Pneumovax has serious limitations when used to evaluate the protection acquired from Pneumovax immunization against any specific serotype.


1992 ◽  
Vol 4 (2) ◽  
pp. 164-169 ◽  
Author(s):  
M. J. McGinley ◽  
D. L. Todd ◽  
H. T. Hill ◽  
K. B. Platt

The potential of a pseudorabies virus (PRV) nucleocapsid protein (NC)-based enzyme-linked immunosorbent assay (ELISA) as a screening assay for PRV infection in subunit-vaccinated and nonvaccinated pigs was studied. The NC-ELISA compared favorably to a commercial ELISA for detecting PRV infection in nonvaccinated pigs. Virus-specific antibody was first detected by the NC-ELISA between days 14 and 21 in 5 pigs challenged intranasally with 104PFU of virus. Antibody continued to be detected in these pigs through day 42, when the experiment was terminated. The NC-ELISA also detected antibody in 23 of 24 pigs from PRV-infected herds. In contrast, the commercial ELISA detected antibody 1 week earlier than the NC-ELISA in experimentally infected pigs but failed to detect antibody in 3 naturally exposed pigs that were identified by the NC-ELISA. Infection in these animals was confirmed by radioimmunoprecipitation analysis. The potential usefulness of the NC-ELISA for detecting infection in vaccinated pigs was also evaluated. The nucleocapsid-specific antibody responses of 10 PRV envelope glycoprotein subunit-vaccinated pigs were monitored prior to and following nasal exposure to a low dose (1023PFU) of PRV. Sera were collected periodically for 113 days after infection. Nucleocapsid-specific antibody responses measured by the NC-ELISA remained below the positive threshold before challenge but increased dramatically following virus exposure. Maximum ELISA responses were obtained on day 32 postchallenge (p.c.). Mean ELISA responses decreased thereafter but remained well above the positive threshold on day 113 p.c. PRV nucleocapsid protein can be used effectively as antigen in the ELISA for detecting PRV infection in both nonvaccinated and subunit-vaccinated pigs.


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