long-term project pp
TRANSCRIPT
Immune Phenotype of patients long-term after allogeneic Hematopoietic Stem Cell Transplantation (HSCT)
Francesca RossiOncology-Pathology dep.,Huddinge HospitalSupervisor: Dr. Michael UhlinCo-supervisor: Arwen Stikvoort
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Hematopoietic Stem Cell Transplantation (HSCT)
transplantation of multipotent hematopoietic stem cells (HSC) Multipotent: they can become one of several types of cells within a given organ Hematopoietic: they are located in the bone marrow and give rise to all other blood cells
through haematopoiesis
An HSCT can be: Autologous: when the patient’s own HSCs are used for the transplantation Allogeneic: when the stem cells come from another individual
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Procedure
1. Conditioning: administration of chemotherapeutic drugs
2. Transplant: HSC get infused into the recipient’s vein
3. Engraftment: the cells reach the sinuses in the bone barrow
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http://www.mdlingo.com/article/reduced-intensity-conditioning-improves-outcome-of-bone-marrow-engraftment
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Possible reactions after HSCT
GVL: immune-mediated response of remission of the hematological malignancy
Graft failure: when host’s immune cell reject the donor’s cells or when part of the HSCT does not work
GVHD: when donor’s cells attack the host’s tissues
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The closest the compatibility is between donor and recipient,the lower is the risk that complications arise
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AIM
Determine whether the donor and patient’s immune system differed after 10 years since transplantation
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?
HSC
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DON: > 10 years
REC: > 10 years
DrugsDiseaseLifestyle…
DON REC
HSCT
DON/REC
HSC
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Material & Methods (1)
Groups:1. 7 donors and 7 recipients Unpaired
2. 5 donors and 5 recipients Paired (donors and patients are siblings)
HSCTs took place between 1987 and 2003
Samples collected at least 10 years ago
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Material & Methods (2)
Samples: PBMC “ring” removed from the blood and frozen (-180°C)
BD FACS Canto Flow cytometer (42 different markers used)
FlowJo analysing software
Statistics: IBM – SPSS Statistics, version 23 GraphPad Prism, version 5.00 P values <0.05
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Unpaired: Mann-Whitney (box plots) Paired: Wilcoxon (paired scatter plots)
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Results (1)
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In a healthy person:
CD4+ > CD8+
Previous reports: shortly after transplantation:
CD8+ > CD4+
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Results (2)
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CD95+ recipients > donors
CD95+ ➡️ CD8+ > CD4+ T cells
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Results (3)
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CD95+ ➡️ CD8+ > CD4+ T cells
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CCR7 – CD45RO
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CD45RO+
CCR7+
TN TCM
TEMTTD
TN: naïve T cells express CCR7 marker (lymph nodes)
TCM: cells that guard against infections
TEM: T cells entered in contact with antigens and are ready to enter the bloodstream to defend the body
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Results (4)
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TEMs ➡️ patients > donors in T cells and CD4+ T cells (A and B) Transplantation increases immune system reaction: mature memory T cells
TNs ➡️ donors > patients in CD4+ T cells (C) Less T naïve cells in patients after HSCT
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Results (5)
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CD45RO+ memory Treg ➡️ patients > donors.
Patients after transplantation: lower levels of naïve T cells
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Results (6)
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PD-1: exhaustion marker ➡️ patients > donors
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To summarize…
CD4+/CD8+ shifted
Higher levels of CD95+ in patients (in both groups)
Higher levels of TEM cells in patients and higher levels of TN in donors
Increased levels of CD45RO+ in Treg memory cells in patients
Higher amounts of PD-1 exhaustion marker in patients’ senescent cells
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Considerations & Conclusions
Aspects that may influence immune system cell’s reconstitution: Quality/quantity of mononuclear cells transplanted Genetic disparity/compatibility between donor and recipient Type of transplantation
Immune system’s cells after HSCT The immune system cells in the patient are its donor’s cells Even after many years in the patient some cell populations seem to show relevant differences
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Thank you!
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Antibodies panel – Flow Cytometry
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Medical data of donors and patients included in the study
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