Large granular lymphocyte leukaemia study at the University Hospital of Donostia.

INTRODUCTION: Large granular lymphocyte (LGL) leukaemia is considered a mature T-cell or natural killer (NK) cell neoplasm, characterised by a clonal proliferation of LGL. AIMS: To analyse the characteristics and to establish (if possible) the prognostic parameters of these patients diagnosed in a single centre: University Hospital of Donostia. METHODS: We retrospectively studied data about 308 patients with LGL leukaemia diagnosed in our centre. RESULTS: The frequency of T-LGL leukaemia and chronic lymphoproliferative disorder of NK cells was 89% and 6.8% respectively, and no aggressive NK-LGL leukaemia was seen in our population. The median age at diagnosis was 65.7 years and male-to-female ratio was 1.08. 59% of our patients were asymptomatic at the time of diagnosis. Most patients presented lymphocytosis and 63.6% more than 20% LGLs in the peripheral blood count, but it has to be taken into account that these results may be influenced by the selection bias of our study, as we recognised these patients as 'alarms of the laboratory analysers'. Neutropenia was the most common cytopenia, and autoimmune disorders were described in 16.5% of the patients. Only 12 patients (3.9%) required treatment, a much lower percentage that the one reported in the literature, and this is consistent with the fact that patients were less symptomatic than in other series, as we expected. The 5-year and 15-year overall survival was 92% and 87%, respectively. CONCLUSIONS: Our patients may represent the even more benign end of the spectrum of clonal T LGL and NK proliferations.

Significance of H63D homozygosity in a Basque population with hemochromatosis.

BACKGROUND: The significance of H63D homozygosity remains uncertain, although it is associated with a tendency for patients to develop iron overload. AIMS: To study the prevalence of homozygotic H63D mutation in patients with phenotypic hemochromatosis (PH) and to compare the results with those of the general population and with patients with porphyria cutanea tarda (PCT) in the Basque Country, Spain. A secondary aim was to evaluate the differences in phenotypic expression and liver injury according to different genotypes in the PH cohort. METHODS: Mutations of the HFE gene were obtained by polymerase chain reaction (PCR). Forty consecutive patients diagnosed with PH, 116 controls and 54 patients with PCT were included in the study. We performed liver biopsies, measured liver iron concentration (LIC), by atomic spectrophotometry, serum ferritin and transferrin saturation, and compared the histology according to the genotype. RESULTS: The H63D homozygote mutation was identified in 7.76% of the control group, in 7.50% of the PH group, and in 11.11% of patients with PCT (P > 0.05). The C282Y/C282Y mutation was present in 50% of patients with PH, and LIC was identified in 15/20. The LIC in C282Y/C282Y patients was higher than in H63D/H63D patients (P = 0.26), while H63D homozygosis caused greater iron overload in PH patients than other genotypes. All the C282Y/C282Y genotype patients had elevated serum ferritin and transferrin saturation. The H63D homozygotes had high ferritin, but two out of three had normal transferrin saturation. Six of the eight patients with high-grade fibrosis and genetic study results were found to be C282Y/C282Y. CONCLUSIONS: The prevalence of H63D mutation in patients with PH in our region does not differ from that of the general Basque population.

Regulatory T cells in patients with inflammatory bowel diseases treated with adacolumn granulocytapheresis.

AIM: To investigate if the clinical efficacy of granulocytes and monocytes by adsorption (GMA) is associated with an increased frequency of peripheral regulatory T cells (Tregs), as these cells have proven to be successful in suppressing inflammatory bowel disease (IBD) in animal models. METHODS: We report four cases of corticosteroid-dependent ulcerative colitis (UC) and two Crohn's disease (CD) cases with severe cutaneous lesions who received GMA therapy. The frequency of CD4+ CD25(high) (Tregs) in peripheral blood was analyzed by flow cytometry and the expression of FoxP3 and TGF beta in purified CD4+ T cells was determined by real time PCR prior to and one month after the last apheresis session, and at the time of endoscopic and clinical assessing. RESULTS: Increased expression of Fox P3 mRNA was found in all five patients who responded to cytapheresis with remission of clinical symptoms, mucosal inflammation and cutaneous lesions, and an increased frequency of circulating Tregs was found in four patients. These changes were not observed in the patient with UC who did no respond to GMA. Variations in TGF-beta (mRNA) did not parallel that of FoxP3 mRNA. CONCLUSION: The clinical efficacy of GMA on IBD and related extra intestinal manifestations was associated with an expansion of circulating CD4+ CD25+ Tregs and higher expression of FoxP3 in CD4+ T cells. Accordingly, an elevated CD4+ CD25+ FoxP3 may be a valuable index of remission in patients with IBD and other chronic relapsing-remitting inflammatory conditions during treatment with GMA.

The impaired response of NK cells from HIV-infected progressor patients to A-class CpG oligodeoxynucleotides is largely dependent of a decreased production of IL-12.

We have reported that NK cells from HIV-infected progressors showed a markedly lower IFN-gamma production in response to an A-class CpG oligodeoxynucleotide as compared to LTNP subjects and healthy HIV-negative individuals. This functional defect was not related to the number of circulating plasmacytoid dendritic cells, nor to the alpha-interferon secreted. In contrast, defective response correlated negatively with the frequency of myeloid dendritic cells. Furthermore, peripheral blood mononuclear cells from LTNPs as well as those from some healthy HIV-negative donors secreted large amounts of IL-12 in unstimulated cultures and in response to the CpG-ODN whereas, HIV-progressor cells showed impaired responses and low level of spontaneous secretion. The addition of a monoclonal anti-IL-12 reduced the response to the CpG-ODN in a dose-dependent manner. These results suggest that the impaired response of NK cells to the CpG-ODN in HIV-progressors is largely dependent on a decreased production of IL-12.

HIV-infected progressors and long-term non-progressors differ in their capacity to respond to an A-class CpG oligodeoxynucleotide.

We used an A-class CpG oligodeoxynucleotide to explore innate immunity in HIV infection and observed that natural killer cells from progressors showed a markedly lower IFN-gamma production in response to the oligonuclotide as compared with long-term non-progressing subjects and healthy HIV-negative individuals. This functional defect was found in patients who showed a long immunological reduction and in those who had had a recent reduction in their CD4 cell counts.