[Initial study on mechanism of a small non-peptidic CD4 inhibitor J2 in prevention corneal allograft rejection]

Abstract

Objective: To discuss the mechanism of J2 on preventing mice's corneal rejection.

Methods: In experimental study, Balb/c mice had been divided into A, B, C, D, E groups randomly. There were 13 mice in each of Group A, C and E, while 7 in both Group B and D. Group A did no management; Group B, autograft control; Group C, D and E, allograft groups (C57BL/6 were used as donors); Group B and Group C were given placebo; Group D and Group E were treated with orally cyclosporine A (CsA) (10 mg per kilogram of body weight) and J2 (15 mg per kilogram of body weight), respectively. To observe the variation of lymphocyte subgroup of peripheral blood mononuclear cells in different groups by flow cytometer analysis every week after corneal transplantation. DTH assay had been done at week 3, cytokines including interleukin-2 (IL-2), IL-10, interferon-gamma (IFN-gamma), secreting level of mouse spleen cells detected by ELLISPOT assay on day 18 after corneal transplantation. To apply single factor analysis of variance, single factor analysis of variance for repeat measured data and so on to analyze the data.

Results: The results of flow cytometer analysis showed: lymphocyte subgroup of peripheral blood mononuclear cells had not proliferated specifically in Group E, compared to Group B and Group D (for CD3(+)CD4(+)T lymphocyte, E-B, P = 0.776, E-D, P = 0.606; for CD3(+)CD8(+)T lymphocyte, E-B, P = 0.941, E-D, P = 0.482). DTH assay showed low reaction to both donor's and third strain mouse spleen cells in Group E (F = 1.00, P = 0.422). The results of ELLISPOT assay indicated: the spot level of IL-2, IFN-gamma in Group E raised slightly compared to Group A, but compared to Group B, there was no statistically significant difference (for IL-2, P = 0.832;for IFN-gamma, P = 0.356). The spot level of IL-10 between all groups had no statistically significant difference (F = 2.911, P = 0.240).

Conclusion: J2 could block up the process of antigen presentation by inhibiting CD4/major histocompatibility complex-II (MHC-II) moleculeonania, while secreting IL-2, IL-10 and IFN-gamma did not have specificity change.