Predominant Th1 cell infiltration in acute rejection episodes of human kidney grafts

Abstract

T-cells and their cytokines are thought to play a major role in the genesis of cellular infiltration and rejection in human kidney allografts. Production of Th1 (IFN-γ) and Th2-type (IL-4 and IL-5) cytokines was assessed in a large series of T-cell clones, derived from core biopsies of kidney grafts in 10 patients with acute interstitial grade I/II rejection (AIR), 6 patients with a histology of 'borderline rejection' (BLR) and 3 with cyclosporine A (CsA) toxicity, all receiving standard maintenance immunosuppression. Biopsies were pre-cultured in IL-2 in order to preferentially expand T-cells activated in vivo, and T-cell blasts were cloned with phytohemagglutinin (PHA) and IL-2 using a highly efficient (23 to 98%) cloning technique. A total of 483 T-cell clones obtained from AIR episodes were compared with 346 and 132 clones derived from patients with BLR episodes and CsA toxicity, respectively. In two series of 22 AIR and 77 BLR T-cell clones, alloreactivity against donor cells was shown by 25 and 14% of CD8+ and 21 and 4% of CD4+ clones, respectively. When stimulated by donor-derived EBV B-cells, all these alloreactive clones produced IFN-γ, but not IL-4 or IL-5 (Th1 clones). Upon stimulation with PHA, the principal qualitative and quantitative differences between AIR- and BLR-derived T-cell clones were that cells derived from AIR patients: (i) showed significantly higher proportions (80 ± 15 vs. 55 ± 13%) of Th1 clones in their progeny; (ii) included smaller proportions (3 ± 4 vs. 20 ± 17%) of clones incapable of producing IFN-γ, IL-4 or IL-5 ('null' clones); and (iii) produced significantly higher quantities of IFN-γ (100 ± 50 vs. 36 ± 7 U/106 cells/ml), these quantities also being significantly correlated (r = 0.83) with the degree of interstitial graft infiltration (item 'i' in the Banff histological grading). The clones derived from CsA toxicity biopsies exhibited a pattern very similar to that found in BLR cases. These data lead us to conclude that the powerful inflammatory response elicited,in acute rejection of a kidney graft recruits and activates both allospecific and non-specific Th1 effector cells, which are primed to high IFN-γ production. Our results also suggest that IFN-γ could contribute, at least in part, to the degree of graft infiltration and to the severity of the rejection episode.