Study of the Function of Immune System in Patients Undergoing Allogeneic Stem Cell Transplantation
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 入组人数
- 177
- 试验地点
- 1
- 主要终点
- The primary endpoint of study is detection of circulating cells by flow cytometry before and after Allogeneic Stem Cell Transplantation
研究概览
简要总结
Graft versus Host disease ( GVHD) is one of the major complications of Allogeneic Stem Cell Transplantation. Acute GVHD develops early ( within 2to 3 months) after transplantation and is the leading cause of death of transplanted patients. The pathogenesis of Chronic GVHD is still little known. Chronic GVHD is caused by donor T lymphocytes, but we have no precise knowledge on the participation of specific subsets of immune system cells to chronic GVHD. In general, chronic GVHD is associated with an increase in the number of T effector lymphocytes, both helper type 2 and cytotoxic.
Recently, also antigen presenting cells (APCs) have been implicated in pathogenesis of chronic GVHD in studies performed on animal models. T lymphocyte responses that characterize chronic GVHD require that recipient antigens are submitted by APCs which originate from the donor's HSC ( Hematopoietic Stem Cells) APCs are heterogeneous population that includes dendritic cells (DCs) ,monocytes, activated B lymphocytes and CD34+ cell subpopulations. These cells can be identified by cytometry.
The data about APCs role in chronic GVHD are preliminary and often discordant. Seemingly, there isn't correlation between circulating APCs number and risk of cGVHD. However, recent data of our group show that patients with cGVHD could have higher number of monocytes in bone marrow than transplanted patients without cGVHD.
The aim of study is to measure the number of circulating immune cells in the PB (peripherical blood) before and after Allogeneic Hematopoietic Stem Cell Transplantation by flow cytometry.
详细描述
Study Endpoints
The primary endpoint of study is detection of circulating cells by flow cytometry before and after Allogeneic Stem Cell Transplantation. We analyze the following cells: Myeloid Dendritic Cells, Plasmacytoid Dendritic cells, CD16+ Dendritic cells, CD14+ monocytes, CD14+/CD16+ monocytes, total T lymphocytes, total T helper lymphocytes, central memory, naive, effector memory and effector T helper lymphocytes, total cytotoxic lymphocytes, central memory, naive, effector memory and effector T cytotoxic lymphocytes, CD16+/CD56+ NK cells, B lymphocytes, T regulatory lymphocytes.
The secondary endpoints are:
- Determination of the serum concentration of Cytokines (TNFα, IFNγ, IL-4) and Chemokines ( MIP and MCP group).
- Determination of autoantibodies in the serum. In the first phase, screening analysis is performed by searching anti nuclear antibodies (ANA) and antibodies anti-ENA (Extractable Nuclear Antigens). Positive results are subjected to second level analysis. Next, we are proceed to the analysis of different Organ-Specific Autoantibodies, previously associated with chronic GVHD, like liver antibodies (SMA, AMA, SLA), anti-cardiolipin and beta2 microglobulin antibodies, Antibodies against thyroid and gastric parietal cells.
- Study of CD86, CXCR4, CCR2, CCR5, CD11a and CD49d molecules expression in Monocytes and Dendritic Cells.
- Study of TNF and IL-12 production by purified monocytes
- Study of Allostimulatory activity of purified monocytes against Allogeneic CD4+ T cells.
- Study of antigen capture and processing by phagocytosis, macropinocytosis and endocytosis mediated by purified monocytes receptors.
- Study of CD134 (0X40L), CD154 (CD40L) molecules expression, study of differentiation molecules type1 and type 2 T helper cells and T regulatory Cells associated, study of molecules Cellular Death associated.
- Study of Cytokines proliferation and production (including IL-2, IL-5, IFNγ and IL-10) and release of perforin and granzyme after stimulation with mitogens.
- Study of antigen-specific T lymphocytes response by pentamers for detection of HY antigen by ELISPOT assay. Recipient's cells are collected and stored before the transplantation
- PCR study of micro (mi) RNA in patient's serum and cells.
- PCR and Microarray study of gene expression profile of transplanted monocytes
Study design
研究设计
- 研究类型
- Observational
- 观察模型
- Cohort
- 时间视角
- Prospective
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •All patients undergoing Allogeneic hemapoietic Stem Cell Transplantation
排除标准
- •Patient not undergoing allogeneic hematopoietic stem cell transplantation
结局指标
主要结局
The primary endpoint of study is detection of circulating cells by flow cytometry before and after Allogeneic Stem Cell Transplantation
时间窗: Patient's PB samples are harvested : on entering the department;1 month after transplantation ; 3, 6, 9,12 months after transplantation
We analyze the following cells:Myeloid Dendritic Cells, Plasmacytoid Dendritic cells, CD16+ Dendritic cells, CD14+ monocytes, CD14+/CD16+ monocytes, total T lymphocytes, total T helper lymphocytes, central memory, naive, effector memory and effector T helper lymphocytes, total cytotoxic lymphocytes, central memory, naive, effector memory and effector cytotoxic lymphocytes, CD16+/CD56+ NK cells, B lymphocytes, T regulatory lymphocytes.
次要结局
未报告次要终点
研究者
Mario Arpinati
MD
IRCCS Azienda Ospedaliero-Universitaria di Bologna