Monitoring T helper cell reconstitution after autologous transplantation of highly purified CD34+ cells in severe autoimmune disease

Journal/Book: Z Rheumatol 1998; 57 Suppl. 1: 75 (P 120). 1998;

Abstract: 1Deutsches Rheuma-Forschungszentrum Berlin High dose ablative chemotherapy followed by autologous transplantation of highly purified CD34+ cells is currently being evaluated as a new strategy for the treatment of severe autoimmune disease. We report here on a patient who received highly purified autologous CD34+ cells for severe relapsing polychronditis. Performing multiparameter flow cytometry we analysed the expression of multiple markers for T helper cell subsets (CD31 CD95 CD62L CD45RA CD45RO CD27 and CD28) on peripheral CD4+ T helper cells highly purified with magnetic cells sorting (MACS) during the immunreconstitution phase and compared it to the pre-tranplantation status. Before mobilisation and without disease activity T helper cell subsets were normal. With disease activity frequency of CD27- T helper cells increased. After mobilisation with G-CSF disease activity reocurred and CD45RO/CD45RA ratio increased. After CD34 selection T cells were below detection limit. Recovery of neutrophiles and platelets were rapid (D12 D8 respectively). Lymphocytes however slowly recovered and absolute counts of CD4+ Th cells were below detection after 4 weeks and low after 8 weeks. Interestingly detectable peripheral blood T helper cells 8 weeks after transplantation lack CD45RA expression and consist exclusively of CD45RO+ memory/effector T helper cells. Among these high frequencies of presumably freshly activated HLA-DR expressing T helper cells and effector T helper cells lacking CD27 expression could be detected. The appearance of these specialised memory/effector T helper cells correlated with atypical pneumonia and may not be a hint for persisting activity of autoimmune disease. Thus highly efficient isolation of CD4+ T helper cells combined with multiparameter flow cytometry allows the rapid and sensitiv detection and analysis of multiple T helper cell subsets even in situations where frequencies of CD4+ T helper cells among peripheral blood cells are extremely low. ... le

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