The Expression Patterns of CD44 and CD45RB on Peripheral Blood T Lymphocytes in the Rejection of Allogeneic Skin Transplantation in Mice.
- Author:
Jae Seok YANG
1
;
Curie AHN
;
Hee Kyeong JUNG
;
Eun Kyeong KIM
;
Jae Young KIM
;
Ki Won KIM
;
Dae Yeon HWANG
;
Jieun OH
;
Yoon Kyu OH
;
Kook Hwan OH
;
Sang Joon KIM
;
Joong Gon KIM
;
Yon Su KIM
;
Jin Suk HAN
;
Suhnggwon KIM
;
Jung Sang LEE
Author Information
1. Department of Nephrology, College of Medicine, Seoul National University, Korea.
- Publication Type:Original Article
- Keywords:
Transplantation rejection;
Peripheral blood T lymphocyte;
CD44
- MeSH:
Animals;
Antibodies, Monoclonal;
Antigens, Surface;
Biopsy;
Diagnosis;
Flow Cytometry;
Follow-Up Studies;
Graft Rejection;
Lymphocytes;
Mass Screening;
Mice*;
Skin Transplantation*;
Skin*;
T-Lymphocytes*;
Tail
- From:The Journal of the Korean Society for Transplantation
2000;14(1):9-22
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: Until now, the rejection was diagnosed through a biopsy, but this method of diagnosis reflected the advanced tissue damage of the transplanted organ and contained the innate problem of being invasive. Activation of T lymphocytes, which occurs before the overt tissue damage has a pivotal role in rejection. In relation, our research attempted to evaluate the viability of analyzing the surface antigens of the peripheral blood activated T lymphocytes in mice after skin transplantation as a noninvasive and early diagnostic tool for diagnosis of rejection. METHODS: After the mouse's skin was transplanted, the expression patterns of activated T lymphocyte markers, CD44 and CD45RB were analyzed along with T lymphocyte markers, CD3, CD4, and CD8 using flow cytometry. The skins from the tails of allogeneic Balb/c (H2(d)) mice and syngeneic C57BL/6J mice were transplanted to C57BL/6J (H2(b)) mice as test and control groups, respectively. Peripheral blood, which was sampled from the tail every other day from day 3 to day 15 was stained with anti-CD44 (or CD45RB), anti-CD4 (or CD8) and anti-CD3 monoclonal antibodies simultaneously, and analyzed by 3-color FACS. Repeated ANOVA test and Mann-Whitey test were used to analyze the differences between the expression patterns of peripheral blood T lymphocyte surface antigen in the control and test groups (SPSS 8.0). RESULTS: Rejection occurred only in the test group from day 8 to day 13 (median: day 10). Although the proportions of CD3(+)lymphocytes (CD3(+)%), CD4(+)lymphocytes (CD4(+) %), and CD8 lymphocytes (CD8(+)%) showed no difference between the control and test groups, the total number of peripheral blood lymphocytes and the number of CD3(+)lymphocytes (CD3(+)) and CD8(+)lymphocytes (CD8(+)) decreased more sharply in the control group after day 7. The proportion and the number of CD44 CD3(+)lymphocytes, CD44 CD4(+)lymphocytes, and CD44(+) CD4(+) CD3(+)lymphocytes began to increase after day 7, to peak on day 11, and then to decrease, showing a significant difference from those of the control group. The proportion and number of CD44(+) CD3(+)lymphocytes, in particular, showed the most significant difference among these significant markers. The proportion and number of CD44(+) CD8(+) lymphocytes and CD44(+) CD8(+) CD3(+)lymphocytes showed similar trends to those of CD44(+) CD3(+) or CD44(+) CD4(+), but the differences between the subset proportions in control and test groups were statistically insignificant. No significant difference was observed in any subsets of the CD45RB antigen. CONCLUSION: CD44(+)CD3(+) lymphocytes representing activated T lymphocytes increased significantly compared to the control group during the rejection period of skin transplantation. The analysis of the expression patterns of surface antigen CD44 on peripheral blood T lymphocytes using flow cytometry is sensitive, safe, easily repeatable, and controllable, and, therefore, can be considered a promising tool for the diagnosis of rejection. However, the clear change in CD44 occurred between day 9 and day 13, when rejection was observed grossly. Therefore, it is regarded more useful as a screening test or follow-up indicator rather than as an early diagnostic tool.
