Flow cytometry panels for immunophenotyping dairy cattle peripheral blood leukocytes

Casaro, Segundo; Marrero, Marcela G.; Madrid, Darling Melany C; Prim, Jessica G.; Nelson, Corwin D.; Galvão, Klibs N.; Laporta, Jimena; Driver, John P.

doi:10.1016/j.vetimm.2022.110417

Cited by 3 publications

(1 citation statement)

References 19 publications

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“…In a study of Norwegian Red calves, substantial changes in leukocyte cell type absolute numbers, lymphocyte subpopulation proportions, and neutrophil functions occurred in the first 5–8 weeks of life with ongoing changes up to 6 months of age [ 36 ]. More recently, 30-day old Holstein calves were shown to have distinctly different lymphocyte subpopulation proportions as compared to adults including fewer B cells and comparatively more γδ T cells [ 37 ]. Furthermore, monocyte subpopulations have been found to be phenotypically and functionally distinct between 10-day old and 18–24 month old Holstein-Friesian cattle both at baseline and following an in vitro Neospora caninum challenge [ 38 ].…”

Section: Discussionmentioning

confidence: 99%

Differential gene expression in peripheral leukocytes of pre-weaned Holstein heifer calves with respiratory disease

Elder,

Hinnant,

Mandella

et al. 2023

PLoS ONE

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Bovine respiratory disease (BRD) is a leading cause of calf morbidity and mortality, and prevalence remains high despite current management practices. Differential gene expression (DGE) provides detailed insight into individual immune responses and can illuminate enriched pathways and biomarkers that contribute to disease susceptibility and outcomes. The aims of this study were to investigate differences in peripheral leukocyte gene expression in Holstein preweaned heifer calves 1) with and without BRD, and 2) across weeks of age. Calves were enrolled for this short-term longitudinal study on two commercial dairies in Washington State. Calves were assessed every two weeks throughout the pre-weaning period using clinical respiratory scoring (CRS) and thoracic ultrasonography (TUS), and blood samples were collected. Calves were selected that were either healthy (n = 10) or had BRD diagnosed by CRS (n = 7), TUS (n = 6), or both (n = 6) in weeks 5 or 7 of life). Three consecutive time point samples were analyzed for each BRD calf consisting of PRE, ONSET, and POST samples. Nineteen genes of interest were selected based on previous gene expression studies in cattle: ALOX15, BPI, CATHL6, CXCL8, DHX58, GZMB, HPGD, IFNG, IL17D, IL1R2, ISG15, LCN2, LIF, MX1, OAS2, PGLYRP1, S100A8, SELP, and TNF. Comparisons were made between age and disease time point matched BRD and healthy calves as well as between calf weeks of age. No DGE was observed between diseased and healthy calves; however, DGE was observed between calf weeks of age regardless of disease state. Developmental differences in leukocyte gene expression, phenotype, and functionality make pre-weaned calves immunologically distinct from mature cattle, and early life shifts in calf leukocyte populations likely contribute to the age-related gene expression differences we observed. Age overshadows disease impacts to influence gene expression in young calves, and immune development progresses upon a common trajectory regardless of disease during the preweaning period.

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Section: Discussionmentioning

confidence: 99%

Differential gene expression in peripheral leukocytes of pre-weaned Holstein heifer calves with respiratory disease

Elder,

Hinnant,

Mandella

et al. 2023

PLoS ONE

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Unraveling the immune and metabolic changes associated with metritis in dairy cows

Casaro,

Prim,

Gonzalez

et al. 2023

Journal of Dairy Science

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Investigating the IgM and IgG B Cell Receptor Repertoires and Expression of Ultralong Complementarity Determining Region 3 in Colostrum and Blood from Holstein-Friesian Cows at Calving

Altvater-Hughes,

Hodgins,

Hodgins

et al. 2024

Animals

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In cattle, colostral maternal immunoglobulins and lymphocytes transfer across the neonate’s intestinal epithelium to provide protection against pathogens. This study aimed to compare repertoires of B cell populations in blood and colostrum in cows for the first time, with an emphasis on ultralong complementarity determining region 3 (CDR3, ≥40 amino acids). Blood mononuclear cells (BMCs, n= 7) and colostral cells (n = 7) were isolated from Holstein-Friesian dairy cows. Magnetic-activated cell sorting was used to capture IgM and IgG B cells from BMCs. Colostral cells were harvested by centrifugation. RNA was extracted and cDNA was produced; IgM and IgG transcripts were amplified using polymerase chain reactions. Amplicons were sequenced using the Nanopore Native barcoding kit 24 V14 and MinION with R10.4 flow cells. In colostrum, there was a significantly greater percentage of IgM B cells with ultralong CDR3s (8.09% ± 1.73 standard error of the mean) compared to blood (4.22% ± 0.70, p = 0.05). There was a significantly greater percentage of IgG B cells in colostrum with ultralong CDR3s (12.98% ± 1.98) compared to blood (6.61% ± 1.11, p = 0.05). A higher percentage of IgM and IgG B cells with ultralong CDR3s in colostrum may be indicative of a potential role in protecting the neonate.

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Flow cytometry panels for immunophenotyping dairy cattle peripheral blood leukocytes

Cited by 3 publications

References 19 publications

Differential gene expression in peripheral leukocytes of pre-weaned Holstein heifer calves with respiratory disease

Differential gene expression in peripheral leukocytes of pre-weaned Holstein heifer calves with respiratory disease

Unraveling the immune and metabolic changes associated with metritis in dairy cows

Investigating the IgM and IgG B Cell Receptor Repertoires and Expression of Ultralong Complementarity Determining Region 3 in Colostrum and Blood from Holstein-Friesian Cows at Calving

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