Immune Development: Role of Milk Cell Transfer and Recipient Gender

免疫发育：乳细胞移植和受体性别的作用

• 批准号: 8206468
• 负责人: AMEAE M WALKER
• 金额: $ 32万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8206468
• 关键词: Adult; Adult Children; Affect; Antibodies; Antigens; B-Lymphocytes; Biological; CD4 Positive T Lymphocytes; CD8B1 gene; Candida albicans; Cells; Child; Complex; Contact hypersensitivity; Delayed Hypersensitivity; Development; Discipline; Discipline of Nursing; Disease; Endocrine; Endocrinology; Epithelial; Epithelial Cells; Equilibrium; Female; Fluorescein; Fostering; Gender; Gene Expression; Genes; Genotype; Green Fluorescent Proteins; Growth Factor; Helper-Inducer T-Lymphocyte; Home environment; Hormones; Human Milk; IL17 gene; Immune; Immune System Diseases; Immune response; Immune system; Immunity; Immunization; Immunohistochemistry; Immunology; Indium; Infant; Interleukin-10; Isothiocyanates; Knowledge; Lactation; Lead; Life; Liquid substance; Male Nurses; Mammary gland; Mediating; Milk; Modeling; Mothers; Mus; Neonatal; Newborn Infant; Nurses; Nutrient; Physiological; Ploidies; Predisposition; Procedures; Process; Production; Prolactin; Public Health; Rag1 Mouse; Regulation; Regulatory T-Lymphocyte; Relative (related person); Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Risk; Role; Sex Characteristics; Sex Chromosomes; Small Intestines; Spleen; System; T-Cell Development; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Testis; Thymus Gland; Time; Tissues; Transgenic Organisms; Tuberculin; Work; World Health; base; cell type; central tolerance; congenic; cytokine; immunopathology; macrophage; male; member; migration; mouse model; novel; novel strategies; offspring; protein B; public health relevance; pup; receptor; response; sex; sexual dimorphism; sry Genes; thymocyte

DESCRIPTION (provided by applicant): We have investigated the trans-epithelial transfer of milk immune cells to nursing pups and examined the result of transfer once the recipients were adult. Our novel approach used foster-nursing of non-transgenic mouse pups by green fluorescent protein (GFP) transgenic dams. The fate of GFP+ immune cells in the pups was followed at 1, 2, 3, and 4 weeks by FACS analysis, immunohistochemistry, and RT-PCR. Despite high numbers of GFP+ B cells and macrophages in milk, these cells were not observed undergoing trans-epithelial migration. Eighty percent of the GFP+ cells observed crossing the small intestine of the pup at weeks 1 and 2 could be positively identified as CD8+ or CD4+ T cells. These T cells homed to the spleen and thymus, with maximal accumulation at 3-4 weeks, and disappearance thereafter. Because T cells were transferred, we examined whether sensitizing dams with an antigen that elicits a T cell-mediated delayed-type-hypersensitivity (DTH) response, would result in sensitization of adult offspring. This was not apparent. Instead, nursing by a sensitized dam amplified a subsequent DTH response in adult female offspring (compared to females nursed by a non-sensitized dam) and yet suppressed a subsequent DTH response in males (compared to males nursed by a non-sensitized dam). In other words, there was a long-lived effect of cell transfer on the DTH response that was different depending on the sex of the recipient. From analysis at later time points, it seems unlikely that the transferred cells became long-lived components of the recipient's immune system, but rather that information was transferred from dam to pup. We therefore hypothesize that transferred maternal cells affect T cell development, selection and expansion during neonatal T cell production. The long term objectives of this project are to determine the importance of maternal milk cell transfer to immune responses in offspring, especially as such transfer might relate a) to T cell development in general, b) to sexual dimorphism both in susceptibility to immune disease and the efficacy of immunization, and c) to the possibility of safely enhancing immunity in young children. The specific aims are 1) to further characterize the transfer of immune information from dam to pup relevant to the adult DTH response. In the process to a) test a working model of the mechanism and b) expand the study to include tuberculin as a disease-related antigen of greater significance to world health, 2) to determine whether the modulation of DTH responsivity is the result of a change in the balance of T cell subsets, and 3) using the novel four core genotype mouse model, to determine whether the sex effect is caused by sex chromosome complement or by the testis-determining gene, Sry. A similar physiological nursing approach is proposed to achieve these specific aims, with mechanistic interrogation enhanced by the addition of Rag1-/-, IL-10 null, IL17 receptor A null, or four core genotype mice as foster recipients of the milk. Congenic CD45.1 and CD45.2 dam and pup pairs will also be used. A team of senior investigators with expertise in prolactin endocrinology and endocrine regulation of immune responses (PI), immunopathology (Muller), mucosal immunology and central tolerance (Lo), and the biological basis of sex differences (Arnold) has been assembled to accomplish these aims. All members of the team are highly productive and respected members of their disciplines. PUBLIC HEALTH RELEVANCE: We have shown that cells transferred from mother to offspring during nursing have opposite modulatory effects on the immune systems in adult male and female recipients. It is anticipated that information garnered from the proposed study will be used to the benefit of infants likely to be at risk for T cell-mediated immune diseases, particularly those showing sexual dimorphism. Results may also lead to the development of procedures for safely enhancing immunity in children.

描述（由申请人提供）： 我们研究了乳汁免疫细胞跨上皮转移至哺乳幼犬的情况，并检查了受体成年后的转移结果。我们的新方法使用绿色荧光蛋白（GFP）转基因母鼠寄养非转基因小鼠幼崽。在第 1、2、3 和 4 周时，通过 FACS 分析、免疫组织化学和 RT-PCR 跟踪幼犬中 GFP+ 免疫细胞的命运。尽管牛奶中存在大量 GFP+ B 细胞和巨噬细胞，但未观察到这些细胞进行跨上皮迁移。在第 1 周和第 2 周观察到穿过幼犬小肠的 GFP+ 细胞中 80% 可以被肯定地鉴定为 CD8+ 或 CD4+ T 细胞。这些 T 细胞归巢于脾脏和胸腺，在 3-4 周时达到最大积累，此后消失。由于 T 细胞被转移，我们检查了用引发 T 细胞介导的迟发型超敏反应 (DTH) 反应的抗原致敏母鼠是否会导致成年后代致敏。这并不明显。相反，由致敏的母鼠哺乳增强了成年雌性后代（与由非致敏母鼠喂养的雌性相比）随后的 DTH 反应，但抑制了雄性随后的 DTH 反应（与由非致敏的母鼠喂养的雄性相比）。换句话说，细胞移植对 DTH 反应的长期影响因接受者的性别而异。从后来的分析来看，转移的细胞似乎不太可能成为受体免疫系统的长寿组成部分，而是信息从母鼠转移到了幼鼠身上。因此，我们假设转移的母体细胞会影响新生儿 T 细胞生产过程中 T 细胞的发育、选择和扩增。 该项目的长期目标是确定母体乳细胞转移对后代免疫反应的重要性，特别是因为这种转移可能与a）一般T细胞发育有关，b）与免疫疾病易感性和免疫效果方面的性别二态性有关，c）与安全增强幼儿免疫力的可能性有关。 具体目标是 1) 进一步表征与成年 DTH 反应相关的免疫信息从母鼠到幼鼠的传递。在此过程中，a) 测试该机制的工作模型，b) 扩大研究范围，将结核菌素作为对世界健康更重要的疾病相关抗原，2) 确定 DTH 反应性的调节是否是 T 细胞亚群平衡变化的结果，3) 使用新型四核心基因型小鼠模型，确定性别效应是由性染色体补体引起还是由睾丸决定基因 Sry 引起。提出了类似的生理护理方法来实现这些特定目标，通过添加 Rag1-/-、IL-10 缺失、IL17 受体 A 缺失或四只核心基因型小鼠作为牛奶的寄养受体来增强机械询问。还将使用同类 CD45.1 和 CD45.2 母鼠对。 为了实现这些目标，我们组建了一支由催乳素内分泌学和免疫反应内分泌调节 (PI)、免疫病理学 (Muller)、粘膜免疫学和中枢耐受 (Lo) 以及性别差异生物学基础 (Arnold) 方面专业知识的高级研究人员组成的团队。团队的所有成员都是高产且在各自学科中受人尊敬的成员。 公共卫生相关性： 我们已经证明，在哺乳期间从母亲转移到后代的细胞对成年男性和女性受体的免疫系统具有相反的调节作用。预计从拟议的研究中获得的信息将用于造福可能面临 T 细胞介导的免疫疾病风险的婴儿，特别是那些表现出性别二态性的婴儿。结果还可能导致安全增强儿童免疫力的程序的开发。