Assessment of Multicolor Flow Cytometry Panels to Study Leukocyte Subset Alterations in Water Buffalo (Bubalus bubalis) During BVDV Acute Infection
The identification of cross-reactive monoclonal antibodies (mAbs) that recognize orthologous leukocyte differentiation molecules (LDM) in buffaloes has overcome a major impediment limiting research on the immune response to pathogens and development of vaccines. As reported, two pilot trials were co...
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Format: | Article |
Language: | English |
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Frontiers Media S.A.
2020-10-01
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Series: | Frontiers in Veterinary Science |
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Online Access: | https://www.frontiersin.org/article/10.3389/fvets.2020.574434/full |
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doaj-117c4b6f84d745ad9602241c4e560364 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Francesco Grandoni Alessandra Martucciello Stefano Petrini Roberto Steri Anna Donniacuo Cristina Casciari Maria Carmela Scatà Carlo Grassi Domenico Vecchio Francesco Feliziani Giovanna De Matteis William C. Davis Esterina De Carlo |
spellingShingle |
Francesco Grandoni Alessandra Martucciello Stefano Petrini Roberto Steri Anna Donniacuo Cristina Casciari Maria Carmela Scatà Carlo Grassi Domenico Vecchio Francesco Feliziani Giovanna De Matteis William C. Davis Esterina De Carlo Assessment of Multicolor Flow Cytometry Panels to Study Leukocyte Subset Alterations in Water Buffalo (Bubalus bubalis) During BVDV Acute Infection Frontiers in Veterinary Science flow cytometry monoclonal antibody water buffalo (Bubalus bubalis) bovine viral diarrhoea virus (BVDV) experimental infection |
author_facet |
Francesco Grandoni Alessandra Martucciello Stefano Petrini Roberto Steri Anna Donniacuo Cristina Casciari Maria Carmela Scatà Carlo Grassi Domenico Vecchio Francesco Feliziani Giovanna De Matteis William C. Davis Esterina De Carlo |
author_sort |
Francesco Grandoni |
title |
Assessment of Multicolor Flow Cytometry Panels to Study Leukocyte Subset Alterations in Water Buffalo (Bubalus bubalis) During BVDV Acute Infection |
title_short |
Assessment of Multicolor Flow Cytometry Panels to Study Leukocyte Subset Alterations in Water Buffalo (Bubalus bubalis) During BVDV Acute Infection |
title_full |
Assessment of Multicolor Flow Cytometry Panels to Study Leukocyte Subset Alterations in Water Buffalo (Bubalus bubalis) During BVDV Acute Infection |
title_fullStr |
Assessment of Multicolor Flow Cytometry Panels to Study Leukocyte Subset Alterations in Water Buffalo (Bubalus bubalis) During BVDV Acute Infection |
title_full_unstemmed |
Assessment of Multicolor Flow Cytometry Panels to Study Leukocyte Subset Alterations in Water Buffalo (Bubalus bubalis) During BVDV Acute Infection |
title_sort |
assessment of multicolor flow cytometry panels to study leukocyte subset alterations in water buffalo (bubalus bubalis) during bvdv acute infection |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Veterinary Science |
issn |
2297-1769 |
publishDate |
2020-10-01 |
description |
The identification of cross-reactive monoclonal antibodies (mAbs) that recognize orthologous leukocyte differentiation molecules (LDM) in buffaloes has overcome a major impediment limiting research on the immune response to pathogens and development of vaccines. As reported, two pilot trials were conducted to accomplish two objectives: (1) demonstrate that multiparameter flow cytometry can be conducted equally well in buffalo with mAbs directly and indirectly labeled with fluorochromes in research and (2) flow cytometry can be used to compare and extend studies on diseases of economic importance to buffalo using bovine viral diarrhea virus (BVDV) as a model pathogen. Pregnant buffalo cows were infected with BVDV-1 at 81 (trial 1) and 203 (trial 2) days post artificial insemination and flow cytometric evaluations were performed at 0, 3, 4, and 14 days after infection (dpi). Fluorochrome conjugated mAbs were used in trial 1, and fluorochrome conjugated goat isotype specific anti-mouse antibodies were used to label mAbs in trial 2. Flow cytometric analysis revealed a transient lymphopenia occurs during the 1st days following infection similar to lymphopenia reported in cattle. In particular, significant differences were observed between pre- and post-infection absolute values of T lymphocytes (−56%, P < 0.01). CD21+ B lymphocytes (−65%, P = 0.04), and Natural Killer cells (−72%, P < 0.001). No significant differences were observed in monocytes and neutrophil absolute values, or the CD4:CD8 ratio. Animal health status was followed until 15 days after calving. No clinical signs of infection were observed during the evaluation period, however, animals in trial 1 developed complications later the infection. One cow aborted at 57 days post-infection, the second cow developed a prolapse a day after calving and died. These two animals also showed a more pronounced lymphopenia in comparison with animals infected at 203 days of pregnancy (e.g., −77 vs. −22% T lymphocytes at 3 dpi, respectively). The pilot studies have demonstrated that it is possible to use multicolour multiparameter flow cytometry to study the immune response to pathogens affecting the health of buffalo. |
topic |
flow cytometry monoclonal antibody water buffalo (Bubalus bubalis) bovine viral diarrhoea virus (BVDV) experimental infection |
url |
https://www.frontiersin.org/article/10.3389/fvets.2020.574434/full |
work_keys_str_mv |
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doaj-117c4b6f84d745ad9602241c4e5603642020-11-25T03:37:46ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692020-10-01710.3389/fvets.2020.574434574434Assessment of Multicolor Flow Cytometry Panels to Study Leukocyte Subset Alterations in Water Buffalo (Bubalus bubalis) During BVDV Acute InfectionFrancesco Grandoni0Alessandra Martucciello1Stefano Petrini2Roberto Steri3Anna Donniacuo4Cristina Casciari5Maria Carmela Scatà6Carlo Grassi7Domenico Vecchio8Francesco Feliziani9Giovanna De Matteis10William C. Davis11Esterina De Carlo12CREA—Consiglio per la Ricerca in Agricoltura e l'Analisi dell'Economia Agraria, Centro di ricerca Zootecnia e Acquacoltura (Research Centre for Animal Production and Aquaculture), Monterotondo, ItalyIstituto Zooprofilattico Sperimentale del Mezzogiorno, National Reference Centre for Hygiene and Technologies of Water Buffalo Farming and Productions, Salerno, ItalyIstituto Zooprofilattico Sperimentale Dell'Umbria e delle Marche, Perugia, ItalyCREA—Consiglio per la Ricerca in Agricoltura e l'Analisi dell'Economia Agraria, Centro di ricerca Zootecnia e Acquacoltura (Research Centre for Animal Production and Aquaculture), Monterotondo, ItalyIstituto Zooprofilattico Sperimentale del Mezzogiorno, National Reference Centre for Hygiene and Technologies of Water Buffalo Farming and Productions, Salerno, ItalyIstituto Zooprofilattico Sperimentale Dell'Umbria e delle Marche, Perugia, ItalyCREA—Consiglio per la Ricerca in Agricoltura e l'Analisi dell'Economia Agraria, Centro di ricerca Zootecnia e Acquacoltura (Research Centre for Animal Production and Aquaculture), Monterotondo, ItalyIstituto Zooprofilattico Sperimentale del Mezzogiorno, National Reference Centre for Hygiene and Technologies of Water Buffalo Farming and Productions, Salerno, ItalyIstituto Zooprofilattico Sperimentale del Mezzogiorno, National Reference Centre for Hygiene and Technologies of Water Buffalo Farming and Productions, Salerno, ItalyIstituto Zooprofilattico Sperimentale Dell'Umbria e delle Marche, Perugia, ItalyCREA—Consiglio per la Ricerca in Agricoltura e l'Analisi dell'Economia Agraria, Centro di ricerca Zootecnia e Acquacoltura (Research Centre for Animal Production and Aquaculture), Monterotondo, ItalyDepartment of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, United StatesIstituto Zooprofilattico Sperimentale del Mezzogiorno, National Reference Centre for Hygiene and Technologies of Water Buffalo Farming and Productions, Salerno, ItalyThe identification of cross-reactive monoclonal antibodies (mAbs) that recognize orthologous leukocyte differentiation molecules (LDM) in buffaloes has overcome a major impediment limiting research on the immune response to pathogens and development of vaccines. As reported, two pilot trials were conducted to accomplish two objectives: (1) demonstrate that multiparameter flow cytometry can be conducted equally well in buffalo with mAbs directly and indirectly labeled with fluorochromes in research and (2) flow cytometry can be used to compare and extend studies on diseases of economic importance to buffalo using bovine viral diarrhea virus (BVDV) as a model pathogen. Pregnant buffalo cows were infected with BVDV-1 at 81 (trial 1) and 203 (trial 2) days post artificial insemination and flow cytometric evaluations were performed at 0, 3, 4, and 14 days after infection (dpi). Fluorochrome conjugated mAbs were used in trial 1, and fluorochrome conjugated goat isotype specific anti-mouse antibodies were used to label mAbs in trial 2. Flow cytometric analysis revealed a transient lymphopenia occurs during the 1st days following infection similar to lymphopenia reported in cattle. In particular, significant differences were observed between pre- and post-infection absolute values of T lymphocytes (−56%, P < 0.01). CD21+ B lymphocytes (−65%, P = 0.04), and Natural Killer cells (−72%, P < 0.001). No significant differences were observed in monocytes and neutrophil absolute values, or the CD4:CD8 ratio. Animal health status was followed until 15 days after calving. No clinical signs of infection were observed during the evaluation period, however, animals in trial 1 developed complications later the infection. One cow aborted at 57 days post-infection, the second cow developed a prolapse a day after calving and died. These two animals also showed a more pronounced lymphopenia in comparison with animals infected at 203 days of pregnancy (e.g., −77 vs. −22% T lymphocytes at 3 dpi, respectively). The pilot studies have demonstrated that it is possible to use multicolour multiparameter flow cytometry to study the immune response to pathogens affecting the health of buffalo.https://www.frontiersin.org/article/10.3389/fvets.2020.574434/fullflow cytometrymonoclonal antibodywater buffalo (Bubalus bubalis)bovine viral diarrhoea virus (BVDV)experimental infection |