Comprehensive Study of 3 Nomenclatures to Discriminate CD8 Subsets in Healthy Volunteers and in Kidney Transplant Recipients
1UMR 1064, INSERM, Nantes, France
2ITUN, CHU de Nantes, Nantes, France
3Faculté
de Médecine, Université
de Nantes, Nantes, France.
Meeting: 2015 American Transplant Congress
Abstract number: D23
Keywords: FACS analysis, Kidney transplantation, Lymphocytes, T cells
Session Information
Session Name: Poster Session D: Costimulation and Signaling in Lymphocytes
Session Type: Poster Session
Date: Tuesday, May 5, 2015
Session Time: 5:30pm-6:30pm
Presentation Time: 5:30pm-6:30pm
Location: Exhibit Hall E
Objective. Historical classification to distinguish central memory (CM), naïve, effector memory (EM), and terminally-differentiated effector memory (TEMRA) CD8 T cells is based on CD45RA and CCR7. Alternative classifications rely on CD45RA in combination with CD27 or CD28. However, the scientific justification of using one classification over another has not been clearly addressed. Thus, we aimed to provide a comprehensive understanding of the benefits of each set of markers in normal and in immune-challenged individuals.
Methods. Multicolor flow cytometry was performed on PBMC from 16 healthy volunteers (HV) and 133 kidney transplant recipients (TX) with a stable graft function to characterize the expression of CD3, CD8, CD27, CD28, CD45RA, CCR7, CD57, TBET, Perforin, Granzyme B and IL7R.
Results. CD45RA+CCR7+, CD45RA+CD27+ and CD45RA+CD28+ identify naïve CD8 T cells with similar phenotypic markers (IL7R+, CD57-, Tbet-, Perf-, and GZMB-) in both HV and TX. While TEMRA can be depicted as CD45RA+CCCR7- or CD45RA+CD27- or CD45RA+CD28-, the use of CD27 or CD28 identifies CD8 T cells with a more differentiated phenotype and increased expression of Tbet, CD57 and perforin in HV and TX. While there is a higher frequency of CD45RA-CD27+ CM (HV 34.96±11.63%; TX 20.78±12.87%) as compared to CD45RA-CCR7+ CM (HV 3.19±1.98%; TX 7.76±6.76%), the expression of CCR7 and CD127 was lower in CM identified using CD27 as opposed to those identified using CCR7 in both HV and TX. CD28 can identify two types of EM cells: CD45RA-CD28+ (EM Early) and CD45RA-CD28- (EM Late) in both HV and TX. When compared to CD45RA-CD27- and CD45RA-CCR7- EM, EM Late showed a higher expression of CD57, Tbet, and GZMB. On the other hand, EM Early displayed similar or lower levels of these markers when compared to CD45RA-CD27- and CD45RA-CCR7- EM cells in HV and TX.
Conclusion. By providing a detailed understanding of CD8 subsets, we illustrate the strength of each set of markers in normal settings and in kidney transplant recipients. Combining CD45RA and CD28 allows for extra insight into the effector potential of CD8 subsets by identifying 2 types of EM cells with different characteristics; alternatively CM nomenclature differed on notably in specific markers.
To cite this abstract in AMA style:
Yap M, Tilly G, Giral M, Brouard S, Degauque N. Comprehensive Study of 3 Nomenclatures to Discriminate CD8 Subsets in Healthy Volunteers and in Kidney Transplant Recipients [abstract]. Am J Transplant. 2015; 15 (suppl 3). https://atcmeetingabstracts.com/abstract/comprehensive-study-of-3-nomenclatures-to-discriminate-cd8-subsets-in-healthy-volunteers-and-in-kidney-transplant-recipients/. Accessed November 21, 2024.« Back to 2015 American Transplant Congress