Erf G F, Ramachandran I R
University of Arkansas, Division of Agriculture, Department of Poultry Science, Fayetteville
University of Arkansas, Division of Agriculture, Department of Poultry Science, Fayetteville.
Poult Sci. 2016 Sep 1;95(9):2011-22. doi: 10.3382/ps/pew122. Epub 2016 Apr 14.
Using the response to Mycobacterium butyricum as the test-immune response, the main goal of this study was to demonstrate the suitability of the growing feather (GF) as a dermal test site and window into in vivo cellular/tissue responses (US-Patent 8,216,551). Using M. butyricum immunized chickens, the specific objectives were to: 1) compare the leukocyte infiltration response to intra-dermally injected M. butyricum in GF, wattles, and wing webs; 2) use GF as the test site to monitor leukocyte response profiles to recall antigen in the same individuals; and 3) gain new knowledge regarding the local response to M. butyricum in chickens. For objective 1, chickens were euthanized for tissue collection at 4 to 6, 24, 48, and 72 h after intra-dermal antigen injection. Leukocyte infiltration profiles were determined using immunochemical and conventional histology. Data from this study established the similarities between the cellular response in GF, wattles, and wing webs and uncovered many advantages of working with GF. For objective 2, antigen was injected into multiple GF per individual. GF were collected before and at 0.25, 1, 2, 3, and 7 d post injection and processed for cell population analysis by flow cytometry. Advantages of the approach used in objective 2 included a technically easier, more comprehensive, and more objective leukocyte profile analysis; same-day data acquisition; and, most importantly, easy, minimally invasive sample collection from the same individual throughout the study. Both studies contributed new knowledge regarding the local cutaneous response to M. butyricum in M. butyricum immunized chickens and confirmed the cell-mediated nature of the immune response to M. butyricum (e.g., elevated levels [P < 0.05] of T cells [CD4+ and CD8+], macrophages and MHC class II+-cells on days one to 3 post injection in M. butyricum- compared to PBS-injected tissues). The use of GF as an "in vivo test tube" to monitor local innate and adaptive immune activities will find direct application in vaccine development, as well as in the assessment and optimization of immune system development and function in poultry.
以对丁酸分枝杆菌的反应作为测试免疫反应，本研究的主要目标是证明生长中的羽毛（GF）作为皮肤测试部位以及深入了解体内细胞/组织反应的可行性（美国专利8,216,551）。使用经丁酸分枝杆菌免疫的鸡，具体目标如下：1）比较在GF、肉垂和翼蹼中皮内注射丁酸分枝杆菌后的白细胞浸润反应；2）以GF作为测试部位，监测同一鸡只对回忆抗原的白细胞反应谱；3）获取有关鸡对丁酸分枝杆菌局部反应的新知识。对于目标1，在皮内注射抗原后的4至6小时、24小时、48小时和72小时对鸡实施安乐死以收集组织。使用免疫化学和传统组织学方法确定白细胞浸润谱。本研究的数据确定了GF、肉垂和翼蹼中细胞反应之间的相似性，并发现了使用GF的诸多优势。对于目标2，向每只鸡的多个GF中注射抗原。在注射前以及注射后0.25天、1天、2天、3天和7天收集GF，并通过流式细胞术进行细胞群体分析。目标2中所采用方法的优势包括技术上更简便、更全面且更客观的白细胞谱分析；当日数据采集；以及，最重要的是，在整个研究过程中能轻松、微创地从同一鸡只采集样本。两项研究均为经丁酸分枝杆菌免疫的鸡对丁酸分枝杆菌的局部皮肤反应贡献了新知识，并证实了对丁酸分枝杆菌免疫反应的细胞介导性质（例如，与注射PBS的组织相比，在注射丁酸分枝杆菌后的第1至3天，T细胞[CD4 +和CD8 +]、巨噬细胞和MHC II类阳性细胞水平升高[P < 0.05]）。将GF用作“体内试管”来监测局部先天性和适应性免疫活动将直接应用于疫苗开发，以及家禽免疫系统发育和功能的评估与优化。
