CD47 Regulates Red Blood Cell Alloimmunization in Mice

CD47型 免疫学 红细胞 抗体 男科 卵清蛋白 抗原 流式细胞术 医学 生物
作者
Ryan Philip Jajosky,Connie M. Arthur,Jerry William Lynn Allen,Megan Fuller,Patricia E. Zerra,Cheryl L. Maier,Sean R. Stowell
出处
期刊:Blood [Elsevier BV]
卷期号:134 (Supplement_1): 100-100 被引量:1
标识
DOI:10.1182/blood-2019-131598
摘要

Background: Exposure to red blood cell (RBC) alloantigens during pregnancy or transfusion can lead to the development of alloantibodies and result in transfusion-related complications, including hemolytic transfusion reactions and hemolytic disease of the fetus and newborn. However, the factors that regulate RBC alloimmunization remain incompletely understood. Several studies suggest that alterations in factors that regulate RBC clearance may impact RBC uptake and antigen presentation, directly influencing the likelihood of RBC alloimmunization. To test this, we directly examined the potential role of CD47, a master regulator of RBC removal previously shown to be altered during RBC senescence and cold storage. To accomplish this, we crossed transgenic mice that express the model HOD antigen (a fusion protein consisting of hen egg lysozyme fused to ovalbumin and human Duffy b) with CD47 knock out (KO) mice to generate HOD RBC donors with wild type, heterozygous or homozygous KO levels of CD47 and used these donors to define the impact of CD47 on antibody formation following RBC transfusion. Methods: HOD transgenic mice expressing the HOD antigen exclusively on RBCs were crossed with CD47-/- mice to produce HOD CD47+/- or HOD CD47-/- mice. HOD and CD47 levels were assessed by flow cytometric analysis using anti-HOD and anti-CD47 antibodies. HOD CD47+/+, HOD CD47+/- or HOD CD47-/- RBCs were transfused into C57BL6 recipients, followed by serum collection on days 14 and 28 post transfusion and evaluation of anti-HOD antibodies by flow cytometry crossmatch. To determine the CD4 T cell response to transfusion, TCR transgenics specific to ovalbumin (OTII) were labeled with CFSE, followed by adoptive transfer, transfusion of HOD CD47+/+, HOD CD47+/- or HOD CD47-/- RBCs and evaluation of T cell proliferation, activation and cytokine secretion. Cellular removal of HOD RBCs was determined by flow cytometric examination of CFSE-labeled HOD RBCs. Finally, antigen levels on HOD RBCs was determined by staining cells with anti-HEL antibodies followed by flow cytometric examination. All three groups were subjected to one-way ANOVA analysis with a p value <0.05 considered significant. Results: While HOD CD47+/+ RBCs expressed levels of CD47 comparable to WT RBCs, HOD CD47+/- RBCs exhibited significantly reduced CD47 levels (nearly half that observed on WT HOD RBCs) and HOD CD47-/-RBCs failed to express any detectable CD47 (p < 0.0001). Following transfusion, HOD CD47+/- and HOD CD47-/- RBCs produced significantly higher levels of IgG anti-HOD antibodies than HOD CD47+/+ RBCs on days 14 and 28 post-transfusion (p < 0.001). However, while HOD CD47-/- RBCs displayed increased clearance consistent with the possible enhancement of a CD4 T cell response (p < 0.0001), HOD CD47+/- RBCs failed to exhibit any different in clearance when compared to HOD CD47+/+ RBCs overtime. To examine the potential impact of differences in HOD RBC clearance on CD4 T cell activation, OTII T cell proliferation was evaluated. While HOD CD47-/- RBC transfusion resulted in significantly increased 33D1+ dendritic cell uptake, OTII proliferation, activation and cytokine secretion (p < 0.05), no difference in 33D1+ dendritic cell uptake or T cell response was observed following HOD CD47+/+ RBC or HOD CD47+/- RBC transfusion. Instead, HOD CD47+/- RBCs exhibited enhanced antigen removal, while also displaying an increased ability to activate HEL specific B cells when compared to HOD CD47+/+ or HOD CD47-/- RBC transfusion. Conclusions: These results demonstrate that alterations in CD47, which occur during normal RBC senescence and cold storage, directly influence RBC alloimmunization through different mechanisms depending on the extent of CD47 loss. While complete loss of CD47 results in enhanced RBC clearance, antigen presentation and CD4 T cell activation, reductions in CD47 to half WT levels failed to impact RBC clearance or T cell activation, but instead enhanced antigen specific B cell activation. These results demonstrate that even partial loss of CD47 is capable of significantly enhancing alloantibody formation completely independent of its role in regulating RBC clearance. In doing so, these findings provide novel insight into the role of CD47 as a key regulator of RBC alloimmunization. Disclosures Stowell: Grifols: Honoraria.

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
wu先生完成签到,获得积分10
刚刚
jmn发布了新的文献求助10
刚刚
刚刚
4秒前
4秒前
5秒前
大米完成签到,获得积分10
5秒前
6秒前
6秒前
ty发布了新的文献求助10
8秒前
田様应助不安的小刺猬采纳,获得10
10秒前
damian完成签到,获得积分10
10秒前
JamesPei应助Fengh采纳,获得10
10秒前
WD发布了新的文献求助10
10秒前
善良天抒发布了新的文献求助10
11秒前
量子星尘发布了新的文献求助30
12秒前
12秒前
xiaojinyu发布了新的文献求助10
13秒前
zxh完成签到,获得积分10
13秒前
15秒前
天天快乐应助obsidian_virgo采纳,获得10
15秒前
16秒前
16秒前
大白完成签到,获得积分10
16秒前
Qiancheng完成签到,获得积分10
16秒前
Owen应助刘YF采纳,获得10
17秒前
ccmaxp发布了新的文献求助10
18秒前
18秒前
ZhaoPeng发布了新的文献求助10
19秒前
一袋薯片发布了新的文献求助10
19秒前
19秒前
gcl应助zwk采纳,获得30
19秒前
20秒前
Owen应助Elhsin_Karte采纳,获得10
21秒前
星辰大海应助善良天抒采纳,获得10
22秒前
jmn完成签到,获得积分10
23秒前
苳苳完成签到 ,获得积分10
23秒前
123发布了新的文献求助10
23秒前
涂山苏苏发布了新的文献求助10
23秒前
Fengh发布了新的文献求助10
24秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
Picture Books with Same-sex Parented Families: Unintentional Censorship 700
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
Nucleophilic substitution in azasydnone-modified dinitroanisoles 500
不知道标题是什么 500
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3975543
求助须知:如何正确求助?哪些是违规求助? 3519971
关于积分的说明 11200248
捐赠科研通 3256311
什么是DOI,文献DOI怎么找? 1798213
邀请新用户注册赠送积分活动 877446
科研通“疑难数据库(出版商)”最低求助积分说明 806338