PSVIII-9 Longitudinal characterization of lymphocyte subsets and humoral immune markers in growing Labrador retrievers.

IF 2.9 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Journal of animal science Pub Date : 2025-10-05 DOI:10.1093/jas/skaf300.529

Fiona B Mccracken, Claire L Timlin, Sarah M Dickerson, Jason W Fowler, Patrick M Skaggs, Craig N Coon

{"title":"PSVIII-9 Longitudinal characterization of lymphocyte subsets and humoral immune markers in growing Labrador retrievers.","authors":"Fiona B Mccracken, Claire L Timlin, Sarah M Dickerson, Jason W Fowler, Patrick M Skaggs, Craig N Coon","doi":"10.1093/jas/skaf300.529","DOIUrl":null,"url":null,"abstract":"This study aimed to characterize the development of the immune system from puppyhood to adulthood in Labrador retrievers. Monthly blood samples were collected from 21 puppies (11M/10F) from 1 month to 1 year of age. For flow cytometry, EDTA whole blood was stained with a multi-color panel including LIVE/DEAD lime viability stain, anti-canine CD4 antibody conjugated to super bright 600, anti-canine CD8 antibody conjugated to super bright 702, anti-canine CD3 antibody conjugated to FITC, anti-canine CD21 antibody conjugated to r-PE, anti-canine CD5 antibody conjugated to PerCP-eFluor 710, and super bright staining buffer. Samples were then lysed and analyzed on an Attune NxT flow cytometer. Gates were determined according to unstained samples, and data was recorded as the percentage of lymphocytes, with the exception of CD4+ and CD8+ cells which were recorded as the percent of CD3+ cells. Plasma samples were utilized for D2Dx analysis at 12, 24, 36, and 50 weeks of age. Data was analyzed in SAS using a repeated measures mixed model with fixed effects of age, sex, and age*sex with dog as the repeated subject, and visualized in JMP. As expected, there were significant effects of age, sex, and age*sex on weight (P ≤ 0.003), with puppies growing over the source of the study, and males on average weighing more than females. There were significant age and sex effects (P ≤ 0.024) for the percentage of CD3+ T cells, which increased throughout the study and which was higher in females. There was an age effect (P &lt; 0.001) for the percentage of CD21+ B cells which decreased as the subjects aged. There were age and sex effects (P ≤ 0.043) for CD5dim cells, tentatively identified as natural killer cells, with levels decreasing over time, and males on average having lower levels than females. The percentage of CD3+CD4+ T cells had significant effects of age and age*sex (P ≤ 0.044), with percentages increasing over time. The percentage of CD3+CD8+ T cells had significant age and sex effects (P ≤ 0.022) with percentages decreasing as the subjects aged, and males having overall lower percentages compared to females. The ratio between CD3+CD4+ and CD3+CD8+ cells had a significant age effect (P &lt; 0.001), but unexpectedly increased considerably over the course of the study. D2Dx scores, a measure of non-cellular humoral immunity, increased significantly between 12, 24, and 36 weeks (P &lt; 0.001), but remained stable between week 36 and the end of the study. Overall, these findings provide valuable insights for researchers and clinicians regarding immune system maturation in Labrador retrievers.","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"31 1","pages":""},"PeriodicalIF":2.9000,"publicationDate":"2025-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of animal science","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1093/jas/skaf300.529","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"AGRICULTURE, DAIRY & ANIMAL SCIENCE","Score":null,"Total":0}

引用次数: 0

Abstract

This study aimed to characterize the development of the immune system from puppyhood to adulthood in Labrador retrievers. Monthly blood samples were collected from 21 puppies (11M/10F) from 1 month to 1 year of age. For flow cytometry, EDTA whole blood was stained with a multi-color panel including LIVE/DEAD lime viability stain, anti-canine CD4 antibody conjugated to super bright 600, anti-canine CD8 antibody conjugated to super bright 702, anti-canine CD3 antibody conjugated to FITC, anti-canine CD21 antibody conjugated to r-PE, anti-canine CD5 antibody conjugated to PerCP-eFluor 710, and super bright staining buffer. Samples were then lysed and analyzed on an Attune NxT flow cytometer. Gates were determined according to unstained samples, and data was recorded as the percentage of lymphocytes, with the exception of CD4+ and CD8+ cells which were recorded as the percent of CD3+ cells. Plasma samples were utilized for D2Dx analysis at 12, 24, 36, and 50 weeks of age. Data was analyzed in SAS using a repeated measures mixed model with fixed effects of age, sex, and age*sex with dog as the repeated subject, and visualized in JMP. As expected, there were significant effects of age, sex, and age*sex on weight (P ≤ 0.003), with puppies growing over the source of the study, and males on average weighing more than females. There were significant age and sex effects (P ≤ 0.024) for the percentage of CD3+ T cells, which increased throughout the study and which was higher in females. There was an age effect (P < 0.001) for the percentage of CD21+ B cells which decreased as the subjects aged. There were age and sex effects (P ≤ 0.043) for CD5dim cells, tentatively identified as natural killer cells, with levels decreasing over time, and males on average having lower levels than females. The percentage of CD3+CD4+ T cells had significant effects of age and age*sex (P ≤ 0.044), with percentages increasing over time. The percentage of CD3+CD8+ T cells had significant age and sex effects (P ≤ 0.022) with percentages decreasing as the subjects aged, and males having overall lower percentages compared to females. The ratio between CD3+CD4+ and CD3+CD8+ cells had a significant age effect (P < 0.001), but unexpectedly increased considerably over the course of the study. D2Dx scores, a measure of non-cellular humoral immunity, increased significantly between 12, 24, and 36 weeks (P < 0.001), but remained stable between week 36 and the end of the study. Overall, these findings provide valuable insights for researchers and clinicians regarding immune system maturation in Labrador retrievers.

查看原文本刊更多论文

生长中的拉布拉多猎犬淋巴细胞亚群和体液免疫标记物的PSVIII-9纵向表征。

本研究旨在描述拉布拉多猎犬从幼犬期到成年期免疫系统的发育。从1个月到1岁，每月采集21只幼犬（11M/10F）的血液样本。流式细胞术中，EDTA全血用多色染色板进行染色，包括LIVE/DEAD lime活力染色，抗犬CD4抗体偶联超亮600，抗犬CD8抗体偶联超亮702，抗犬CD3抗体偶联FITC，抗犬CD21抗体偶联r-PE，抗犬CD5抗体偶联PerCP-eFluor 710，以及超亮染色缓冲液。然后在tune NxT流式细胞仪上对样品进行裂解和分析。根据未染色的样品确定gate，数据记录为淋巴细胞的百分比，CD4+和CD8+细胞记录为CD3+细胞的百分比。在12、24、36和50周龄时，血浆样本用于D2Dx分析。SAS采用年龄、性别、年龄*性别固定效应的重复测量混合模型，以狗为重复被试进行数据分析，并在JMP中进行可视化。正如预期的那样，年龄、性别和年龄*性别对体重有显著影响（P≤0.003），幼犬的生长超过了研究来源，雄性的平均体重高于雌性。CD3+ T细胞的百分比存在显著的年龄和性别效应（P≤0.024），在整个研究过程中，CD3+ T细胞的百分比都在增加，其中女性更高。CD21+ B细胞的百分比随着受试者年龄的增长而下降，存在年龄效应（P < 0.001）。CD5dim细胞（初步确定为自然杀伤细胞）存在年龄和性别效应（P≤0.043），随着时间的推移，CD5dim细胞的水平逐渐降低，男性的平均水平低于女性。CD3+CD4+ T细胞百分比受年龄和年龄*性别的影响显著（P≤0.044），随时间增加百分比增加。CD3+CD8+ T细胞的百分比存在显著的年龄和性别效应（P≤0.022），随着受试者年龄的增长，百分比逐渐降低，男性的百分比总体低于女性。CD3+CD4+和CD3+CD8+细胞之间的比率具有显著的年龄效应（P < 0.001），但出乎意料地在研究过程中显著增加。测量非细胞体液免疫的D2Dx评分在12周、24周和36周之间显著增加（P < 0.001），但在第36周和研究结束之间保持稳定。总的来说，这些发现为研究人员和临床医生提供了关于拉布拉多猎犬免疫系统成熟的宝贵见解。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Journal of animal science 农林科学-奶制品与动物科学

CiteScore

4.80

自引率

12.10%

发文量

1589

审稿时长

3 months

期刊介绍： The Journal of Animal Science (JAS) is the premier journal for animal science and serves as the leading source of new knowledge and perspective in this area. JAS publishes more than 500 fully reviewed research articles, invited reviews, technical notes, and letters to the editor each year. Articles published in JAS encompass a broad range of research topics in animal production and fundamental aspects of genetics, nutrition, physiology, and preparation and utilization of animal products. Articles typically report research with beef cattle, companion animals, goats, horses, pigs, and sheep; however, studies involving other farm animals, aquatic and wildlife species, and laboratory animal species that address fundamental questions related to livestock and companion animal biology will be considered for publication.