Experimental animal transplantationImmunobiologyImmunomodulation of Human CD8+ T Cells by Thymoglobulin In Vitro
Section snippets
Purification of CD8+ Cells and Pretreatment With Thymoglobulin
CD8+ cells purified from peripheral blood mononuclear cells collected from volunteers by negative selection were verified using flow cytometry showing >94% CD3+/CD8+ cells. CD8+ cells (2 × 106 cells/mL) were incubated with 10 μg/mL of Thymoglobulin or rabbit isotype immunoglobulin (Ig) on ice for 2 hours before washing 3 times to remove unbound Thymoglobulin or isotype Ig. Cells diluted to 2 × 106 cells/mL were incubated at 37°C with supernates collected at 24, 48, and 72 hours.
Quantification of Transcripts Using Real-Time Quantitative Polymerase Chain Reaction
Real-time
Responses of CD8+ Cell Gene Transcripts After Antibody Treatment
This study evaluated initial responses of CD8+ cells after Thymoglobulin pretreatment for costimulatory and regulatory molecule and cytokine gene transcripts. As shown in Figure 1, RT-PCR demonstrated Thymoglobulin-treated CD8+ cells to up-regulate transcripts for CTLA-4, OX40, GITR, Foxp3, IFN-γ, IL-10, CD25, and IL-2 when compared with isotype Ig-treated cells. The expression for GITR and IL-2 transcripts was down-regulated at 72 hours after incubation.
Cytokine Release From CD8+ Cells After Antibody Treatment
CD8+ cells, pretreated with
Discussion
Thymoglobulin, which has been used as induction therapy to prevent ARE,1 binds multiple lymphocyte surface antigens.2 Lymphocyte depletion has been believed to be the major mechanism for its therapeutic anti-rejection effects. Previous studies have shown effects of Thymoglobulin to modulate human CD4+ cells in vitro.7 This study explored gene transcript expression, cytokine production, and surface costimulatory molecule expression on purified CD8+ cells following pretreatment with Thymoglobulin.
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Basics and Art of Immunosuppression in Liver Transplantation
2024, Journal of Clinical and Experimental HepatologyManagement of Immunosuppression in Liver Transplantation
2017, Clinics in Liver DiseaseCitation Excerpt :ATG and ALG are polyclonal antibodies to multiple T-cell antigens, CD2, CD3, CD4, and CD8. Treatment with them induces opsonization of lymphocytes and subsequent complement-medicated cell lysis, thus eventually leading to lymphocyte depletion.48 OKT3 is a monoclonal antibody that targets CD3 on mature T cells and leads to inactivation and apoptosis of T cells.49
The use and value of procalcitonin in solid organ transplantation
2015, Clinical Transplantation