H Wu1, G Y Zhang, A R Clarkson, J F Knight. 1. Center for Kidney Research, Royal Alexandra Hospital for Children, Westmead, New South Wales, and Renal Unit, Royal Adelaide Hospital, Adelaide, South Australia, Australia. Huilw@nch.edu
Abstract
BACKGROUND: We have previously reported that both alpha beta and gamma delta T cells are involved in the progression of IgA nephropathy (IgAN) to renal failure. To determine whether the T-cells seen in the interstitium represent a generalized inflammatory response or whether they are proliferating oligoclonally in response to a particular antigen, we analyzed the TCR Vbeta gene usage by T cells infiltrating renal biopsies from patients with IgAN. METHODS: Fourteen IgAN patients were divided by clinical criteria into stable and progressive groups (7 in each group). Reverse transcription-polymerase chain reaction (RT-PCR) and cloning were used to characterize the expression of TCR Vbeta families in renal biopsies and in peripheral blood lymphocytes. RESULTS: TCR Vbeta8 was significantly and preferentially expressed in most IgAN kidney biopsies compared with peripheral blood lymphocytes from both IgAN patients and healthy controls (P < 0.001). TCR Vbeta8 expression was more marked in progressive biopsies than in stable biopsies (P < 0.05). Spectratyping of Vbeta8 RT-PCR products from T cells infiltrating the kidney showed an intense spectratype band at the shortest range of amplified CDR3s in the renal biopsies of four patients. Analysis of nucleotide and deduced amino acid sequences of Vbeta8 PCR products derived from intense spectratype bands from these renal biopsies revealed a high concordance across the CDR3 region. A conserved amino acid (leucine) at the first position of the nongermline-encoded nucleotides and diversity (ND) junction of Vbeta8 was found at a frequency of 95% in multiple sequences obtained from the renal biopsies of all four patients examined. CONCLUSIONS: The preferential use of Vbeta8 with marked similarities in the CDR3 region by some renal infiltrating T cells suggests clonal expansion of T cells in the kidneys of some IgAN patients. Conserved amino acids in the TCR CDR3 hypervariable region may contribute to the recognition of a particular antigen or set of antigens.
BACKGROUND: We have previously reported that both alpha beta and gamma delta T cells are involved in the progression of IgA nephropathy (IgAN) to renal failure. To determine whether the T-cells seen in the interstitium represent a generalized inflammatory response or whether they are proliferating oligoclonally in response to a particular antigen, we analyzed the TCR Vbeta gene usage by T cells infiltrating renal biopsies from patients with IgAN. METHODS: Fourteen IgANpatients were divided by clinical criteria into stable and progressive groups (7 in each group). Reverse transcription-polymerase chain reaction (RT-PCR) and cloning were used to characterize the expression of TCR Vbeta families in renal biopsies and in peripheral blood lymphocytes. RESULTS: TCR Vbeta8 was significantly and preferentially expressed in most IgAN kidney biopsies compared with peripheral blood lymphocytes from both IgANpatients and healthy controls (P < 0.001). TCR Vbeta8 expression was more marked in progressive biopsies than in stable biopsies (P < 0.05). Spectratyping of Vbeta8 RT-PCR products from T cells infiltrating the kidney showed an intense spectratype band at the shortest range of amplified CDR3s in the renal biopsies of four patients. Analysis of nucleotide and deduced amino acid sequences of Vbeta8 PCR products derived from intense spectratype bands from these renal biopsies revealed a high concordance across the CDR3 region. A conserved amino acid (leucine) at the first position of the nongermline-encoded nucleotides and diversity (ND) junction of Vbeta8 was found at a frequency of 95% in multiple sequences obtained from the renal biopsies of all four patients examined. CONCLUSIONS: The preferential use of Vbeta8 with marked similarities in the CDR3 region by some renal infiltrating T cells suggests clonal expansion of T cells in the kidneys of some IgANpatients. Conserved amino acids in the TCR CDR3 hypervariable region may contribute to the recognition of a particular antigen or set of antigens.