- Email: [email protected]
Relationship of T cell proliferation and the udder transcriptome with mastitis resistance to Staphlococcus aureus
Abstracts / Veterinary Immunology and Immunopathology 128 (2009) 211–347
261
Relationship of T cell proliferation and the udder transcriptome with mastitis resistance to Staphlococcus aureus
way targets as candidates for new genetic tests of mastitis resistance.
Nicola Hastings 1 , Fiona Young 2 , John Williams 3 , Julie Fitzpatrick 4 , Elizabeth J. Glass 1
doi:10.1016/j.vetimm.2008.10.109
1
Roslin Institute, Roslin, Midlothian EH25 9PS, UK 2 Agriculture Branch, Agri-Food and Biosciences Institute, Hillsborough, County Down, Northern Ireland, BT26 6DR, UK 3 Parco Tecnologico Padano,Via Einstein, Polo Universitario, Lodi 26900, Italy 4 Moredun Reseach Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ, UK Keywords: Mastitis; Resistance; Cattle; Microarray Species: Ruminants Mastitis remains one of the most important diseases in dairy cattle particularly in the western world. It costs the EU alone D 100–200 million/year with economic losses due to reduced milk production, lower milk quality, veterinary treatment and also culling. In addition, as mastitis is an extremely painful disease, there are serious welfare implications. Current control measures are not always effective and developing vaccines targeted at mucosal surfaces such as the udder are proving difficult. Breeding for resistance to mastitis could be an alternative means of control but is not straightforward for a number of reasons. Although it is clear that there is a genetic component accounting for variation in mastitis resistance, it is likely to be multigenic and the trait has low heritability, possibly because mastitis scoring for genetic analysis usually does not take into consideration that mastitis is caused by different pathogens which elicit different host responses. Additionally mastitis is becoming more of an issue for dairy farmers because it has a negative correlation with genetic selection for increased productivity. We propose to take a different approach by taking advantage of the genomic resources now available for cattle to identify key genes and pathways that could ultimately lead to new genetic tests and selectable markers for breeding for mastitis resistance. Earlier studies indicated that measurement of the proliferative response of bovine peripheral blood T cells to formalin-fixed Staphylococcus aureus might be used as a potential mastitis-resistance predictor with high responders potentially predicting greater resistance to S. aureus mastitis than those with a low proliferative response. Our studies in a cattle cross-population showed that the level of proliferation to S. aureus was at least partially genetically determined (h2 = 0.2). We now aim to directly investigate whether young heifers selected on the basis of their T cell response to S. aureus into two groups of high and low responders, do differ in their response to experimental S. aureus challenge in vivo at 6-weeks of lactation. In addition to measuring clinical parameters, we will also transcriptionally profile the cells entering the udder and the udder tissue itself following infection. This project could provide more direct evidence of the predictive ability of the T cell test, and has the potential to reveal relevant gene and path-
Evaluation of Brucella abortus hydrophobic and hydrophilic fractions by avidity-immunoblot for serodiagnosis of bovine brucellosis Ana C.A.M. Pajuaba, Deise A.O. Silva, Dâmaso P. Ribeiro, José R. Mineo ∗ Laboratory of Immunoparasitology, Institute of Biomedical Sciences, Federal University of Uberlândia, Av. Pará 1720, 38400-902 Uberlândia, MG, Brazil Keywords: Brucella abortus; Avidity-immunoblot; Triton X-114; Cattle; LPS E-mail address: [email protected] (J.R. Mineo). Specie: Ruminants Brucellosis is a major zoonosis and has been considered an emerging or re-emerging disease worldwide, particularly leading to abortion and infertility in domestic herds. Diagnosis of brucellosis in cattle is mainly based on serological methods that have used whole cell preparations, sonicated cell extracts or lipopolysaccharide (LPS) enriched fractions. As specificity of these tests is low, alternative antigens have been characterized as potentially useful tools in diagnostic tests for brucellosis. In this context, the Triton X-114 nonionic detergent has been successfully used for extraction of membrane-associated proteins, but no data are available on its use for Brucella abortus antigen extraction. Smooth lipopolysaccharide (S-LPS) and Triton X-114 fractions from B. abortus were analyzed in immunoblot and avidity-immunoblot to identify differences between antigenic markers from S19 vaccinated from non-vaccinated seropositive cows. Four groups with 15 cattle sera each were analyzed: (I) non-vaccinated seropositive cows from Brucella-endemic areas, (II) non-vaccinated seropositive cows from Brucella non-endemic areas, (III) S19 vaccinated heifers, and (IV) non-vaccinated seronegative cows. Classical agglutination tests were used to select the groups of cattle sera. S-LPS immunoblot showed a wide cluster of bands (22–105 kDa) recognized by sera from non-vaccinated cows (groups I and II) while a more restrict cluster (43–58 kDa) was seen in sera from vaccinated heifers (group III). In avidity-immunoblot using S-LPS, no distinct reactivity profile could be identified. Triton X-114 hydrophobic fraction revealed a wide cluster of antigenic bands in contrast to hydrophilic fraction that showed a more clearly defined reactivity profile. Immunoblot using the hydrophilic fraction revealed some immunodominant antigens (30, 35, 43, 57 and 73 kDa) in all three seropositive groups, whereas a significantly lower reactivity was seen for the 69 and 65 kDa bands only in vaccinated heifers (group III). Avidity-immunoblot with the hydrophilic fraction showed significant reactivity impairment for the immunodominant antigenic bands (57, 43 and 35 kDa) recognized by sera of group III, reflecting in
261
Relationship of T cell proliferation and the udder transcriptome with mastitis resistance to Staphlococcus aureus
way targets as candidates for new genetic tests of mastitis resistance.
Nicola Hastings 1 , Fiona Young 2 , John Williams 3 , Julie Fitzpatrick 4 , Elizabeth J. Glass 1
doi:10.1016/j.vetimm.2008.10.109
1
Roslin Institute, Roslin, Midlothian EH25 9PS, UK 2 Agriculture Branch, Agri-Food and Biosciences Institute, Hillsborough, County Down, Northern Ireland, BT26 6DR, UK 3 Parco Tecnologico Padano,Via Einstein, Polo Universitario, Lodi 26900, Italy 4 Moredun Reseach Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ, UK Keywords: Mastitis; Resistance; Cattle; Microarray Species: Ruminants Mastitis remains one of the most important diseases in dairy cattle particularly in the western world. It costs the EU alone D 100–200 million/year with economic losses due to reduced milk production, lower milk quality, veterinary treatment and also culling. In addition, as mastitis is an extremely painful disease, there are serious welfare implications. Current control measures are not always effective and developing vaccines targeted at mucosal surfaces such as the udder are proving difficult. Breeding for resistance to mastitis could be an alternative means of control but is not straightforward for a number of reasons. Although it is clear that there is a genetic component accounting for variation in mastitis resistance, it is likely to be multigenic and the trait has low heritability, possibly because mastitis scoring for genetic analysis usually does not take into consideration that mastitis is caused by different pathogens which elicit different host responses. Additionally mastitis is becoming more of an issue for dairy farmers because it has a negative correlation with genetic selection for increased productivity. We propose to take a different approach by taking advantage of the genomic resources now available for cattle to identify key genes and pathways that could ultimately lead to new genetic tests and selectable markers for breeding for mastitis resistance. Earlier studies indicated that measurement of the proliferative response of bovine peripheral blood T cells to formalin-fixed Staphylococcus aureus might be used as a potential mastitis-resistance predictor with high responders potentially predicting greater resistance to S. aureus mastitis than those with a low proliferative response. Our studies in a cattle cross-population showed that the level of proliferation to S. aureus was at least partially genetically determined (h2 = 0.2). We now aim to directly investigate whether young heifers selected on the basis of their T cell response to S. aureus into two groups of high and low responders, do differ in their response to experimental S. aureus challenge in vivo at 6-weeks of lactation. In addition to measuring clinical parameters, we will also transcriptionally profile the cells entering the udder and the udder tissue itself following infection. This project could provide more direct evidence of the predictive ability of the T cell test, and has the potential to reveal relevant gene and path-
Evaluation of Brucella abortus hydrophobic and hydrophilic fractions by avidity-immunoblot for serodiagnosis of bovine brucellosis Ana C.A.M. Pajuaba, Deise A.O. Silva, Dâmaso P. Ribeiro, José R. Mineo ∗ Laboratory of Immunoparasitology, Institute of Biomedical Sciences, Federal University of Uberlândia, Av. Pará 1720, 38400-902 Uberlândia, MG, Brazil Keywords: Brucella abortus; Avidity-immunoblot; Triton X-114; Cattle; LPS E-mail address: [email protected] (J.R. Mineo). Specie: Ruminants Brucellosis is a major zoonosis and has been considered an emerging or re-emerging disease worldwide, particularly leading to abortion and infertility in domestic herds. Diagnosis of brucellosis in cattle is mainly based on serological methods that have used whole cell preparations, sonicated cell extracts or lipopolysaccharide (LPS) enriched fractions. As specificity of these tests is low, alternative antigens have been characterized as potentially useful tools in diagnostic tests for brucellosis. In this context, the Triton X-114 nonionic detergent has been successfully used for extraction of membrane-associated proteins, but no data are available on its use for Brucella abortus antigen extraction. Smooth lipopolysaccharide (S-LPS) and Triton X-114 fractions from B. abortus were analyzed in immunoblot and avidity-immunoblot to identify differences between antigenic markers from S19 vaccinated from non-vaccinated seropositive cows. Four groups with 15 cattle sera each were analyzed: (I) non-vaccinated seropositive cows from Brucella-endemic areas, (II) non-vaccinated seropositive cows from Brucella non-endemic areas, (III) S19 vaccinated heifers, and (IV) non-vaccinated seronegative cows. Classical agglutination tests were used to select the groups of cattle sera. S-LPS immunoblot showed a wide cluster of bands (22–105 kDa) recognized by sera from non-vaccinated cows (groups I and II) while a more restrict cluster (43–58 kDa) was seen in sera from vaccinated heifers (group III). In avidity-immunoblot using S-LPS, no distinct reactivity profile could be identified. Triton X-114 hydrophobic fraction revealed a wide cluster of antigenic bands in contrast to hydrophilic fraction that showed a more clearly defined reactivity profile. Immunoblot using the hydrophilic fraction revealed some immunodominant antigens (30, 35, 43, 57 and 73 kDa) in all three seropositive groups, whereas a significantly lower reactivity was seen for the 69 and 65 kDa bands only in vaccinated heifers (group III). Avidity-immunoblot with the hydrophilic fraction showed significant reactivity impairment for the immunodominant antigenic bands (57, 43 and 35 kDa) recognized by sera of group III, reflecting in