Effects of extracorporeal photoimmunotherapy on soluble IL-2Ralpha,TNF-RI,and CD8 in patients with steroid-resistant acute graft-versus-host disease

Extracorporeal photoimmunotherapy (ECP) has been successfully used as adjunct treatment for steroid-resistant graft-versus-host disease (GvHD) after allogeneic stem cell transplantation. We serially investigated serum levels of soluble interleukin-2 receptor-alpha (sIL-2Ralpha), soluble tumor necrosis factor receptor I (sTNF-RI), and soluble CD8 (sCD8) in 19 patients with steroid-resistant acute GvHD before and after each ECP treatment. Highest levels of sIL-2Ralpha and sTNF-RI correlated with severe acute GvHD and infections. Despite an immediate sIL-2Ralpha and sTNF-RI decrease after each treatment cycle, a mean surge of sTNF-RI>sIL-2Ralpha during the first three ECP cycles was observed in infections. A delayed surge, i.e., after the third ECP cycle, of sIL-2Ralpha and elevated post-ECP sCD8 levels was observed in patients developing chronic GvHD. While levels of sIL-2Ralpha and sTNF-RI correlate with the severity of acute GvHD and infections during the early ECP treatment period, the recurring increase of post-ECP sCD8 possibly may serve as parameter for developing chronic GvHD.     

Expression of adhesion molecules in human intestinal graft-versus-host disease.

The distribution of three cellular adhesion molecules, ICAM-1, ELAM-1 and VCAM-1, was studied in normal rectal mucosa and in graft-versus-host disease (GvHD) using immunohistological and morphometric techniques. In normal controls, ICAM-1 was demonstrable on endothelial cells and leucocytes within the lamina propria, ELAM-1 on endothelial cells only and VCAM-1 on lamina propria leucocytes, many of which exhibited long dendritic processes surrounding the glands. In GvHD, the enterocytes became positive for ICAM-1 and this was often associated with the presence of intra-epithelial LFA-1+ lymphocytes and macrophages, the latter containing debris of apoptotic cells. The staining was, however, restricted to the luminal membrane of the epithelial cells, raising doubts about the role of ICAM-1 as a ligand for LFA-1 on mucosal leucocytes in rectal GvHD. ELAM-1 expression was increased in GvHD both in terms of the length of positive endothelium and staining intensity. VACM-1 was increased on endothelial cells but not leucocytes in the lamina propria in contrast to our previous findings in cutaneous GvHD where VCAM-1+ dendritic cells were increased and endothelial cells remained negative. Normal patterns of adhesion molecule staining were seen in two biopsies exhibiting no morphological evidence of GvHD, from patients who had strong clinical evidence of the disease, indicating that immunostaining for these molecules is unlikely to be of help in improving the sensitivity of histological diagnosis. However, the possibility that adhesion molecule staining may be useful in improving diagnostic specificity by helping to distinguish GvHD from identical histological changes produced by irradiation and cytotoxic drugs is worthy of further investigation.     

“Assessment of chronic sclerodermoid Graft‐versus‐Host Disease patients,using 20MHz high‐frequency ultrasonography and cutometer methods”

The development of an adverse graft‐versus‐host disease (GvHD) is a major complication of stem cell transplantations, which are widely used to cure increasing number of hematologic malignancies. Patients with chronic GvHD are at risk of joint contractures secondary to sclerodermatous skin changes. Several clinical scores or serologic markers have been used to assess skin sclerosis in scleroderma patients. Evaluation of sclerotic skin changes using biometric tools remains to be challenging. The purpose of this study was to illustrate and exemplify ultrasound measurement and measurement of skin elasticity of five chronic sclerodermoid GvHD patients. There is still a substantial lack of studies using objective and non‐invasive methods helpful in assessment of patients with skin involvement of GvHD. Although ultrasound is not the ideal method, it is worth emphasizing that it is still useful, non‐invasive, and repeatable device in monitoring patients suffering from GvHD. It should also be added, that it seems to be advisable to repeat USG examination at an interval of 3 months after the treatment. In addition, skin echogenicity may be a more sensitive parameter than skin thickness in assessment of cGvHD patients.     

Novel application of lentiviral vectors towards treatment of graft-versus-host disease

Allogeneic transplantation of hematopoietic stem cells and lymphocytes is a curative treatment for malignant and non-malignant disease. However, the primary complication limiting the safety of transplantation is graft-versus-host disease (GvHD), which is mediated by donor T cells. Strategies for pre- and post-transplant manipulation of graft cells are not yet optimal for balancing GvHD severity with beneficial Graft-versus-Leukemia (GvL) effects. Emerging cell fate control gene therapy (CFCGT)-based strategies, such as ‘suicide’ gene therapy for donor T cell regulation, can supplement existing transplantation approaches by providing a safety element to reduce GvHD. Past uses of CFCGT in the clinic have provided proof-of-principle that GvHD can be controlled by such a strategy. However, there exists a need for improved transgene delivery and suicide control systems. Recently, lentiviral vectors (LVs) have emerged as effective gene delivery vehicles for the clinic. Combining lentiviral gene delivery with newer generations of ‘suicide’ systems that possess improved enzyme/prodrug specificities, activities, and reduced immunogenicity, could provide the necessary degree of control required to more successfully manage GvHD. Improving the safety of transplantation through successful CFCGT will serve to expand the potential donor pool and the spectrum of disorders that can be treated by this therapeutic schema.     

Peripheral blood late mixed chimerism in leucocyte subpopulations following allogeneic stem cell transplantation for childhood malignancies: does it matter?

The impact of persistent mixed chimerism (MC) after haematopoietic stem cell transplantation (HSCT) remains unclarified. We investigated the incidence of MC in peripheral blood beyond day +50 after HSCT and its impact on rejection, chronic graft‐versus‐host disease (c‐GvHD) and relapse in 161 children receiving allogeneic HSCT for haematological malignancies. The 1‐year incidence of late MC was 26%. Spontaneous conversion to complete donor chimerism (CC) occurred in 43% of patients as compared to 62% after donor lymphocyte infusions. No graft rejection occurred. The 1‐year incidence of c‐GvHD was 20 ± 7% for MC, and 18 ± 4% for CC patients (P = 0·734). The 3‐year cumulative incidence of relapse (CIR) according to chimerism status at days +50 and +100 was 22 ± 4% for CC patients vs. 22 ± 8% for MC patients (day +50; P = 0·935) and 21 ± 4% vs. 20 ± 7% (day +100; P = 0·907). Three‐year CIRs in patients with persistent MC and patients with CC/limited MC were comparable (8 ± 7% vs. 19 ± 4%; P = 0·960). HSCT for acute leukaemia or myelodysplastic syndrome as secondary malignancies (hazard ratio (HR) 4·7; P = 0·008), for AML (HR 3·0; P = 0·02) and from mismatched donors (HR 3·1; P = 0·03) were independent factors associated with relapse. Our data suggest that late MC neither protects from c‐GvHD nor does it reliably predict impending disease relapse.     

Antibodies to 60, 65 and 70 kDa heat shock proteins in pediatric allogeneic stem cell transplant recipients

Allogeneic SCT remains the only means of cure for many patients with various malignant disorders as well as non-malignant diseases. Infection together with severe aGvHD may result in a significant incidence of transplant-related morbidity and mortality. Current evidence suggests that hSPS represent major immunodominant antigens in many pathogens and therefore might play an important role in the pathogenesis of GvHD. We investigated the levels of total Ig, IgG and IgM isotype antibodies to rh-hsp60, recombinant Mycobacterium bovis hsp65 and stress-inducible rh-hsp70 in sera of pediatric patients undergoing SCT by using ELISA. We studied whether humoral immune responses to hSPS follow transplant-related complications, bacterial and fungal infection. Anti-hsp antibodies were detected in patients' sera before conditioning, over the course of conditioning and all the time post-transplant. We found no correlation between anti-hsp antibodies and the occurrence and severity of GvHD and/or other transplant-related complications like graft failure, hemorrhagic cystitis and capillary leakage syndrome. However, elevated anti-hsp antibodies involving IgM and IgG isotypes were found to be associated with bacterial and fungal infection depending on etiological agents. We demonstrated de novo humoral response to hSPS in a cohort of patients with actual infection caused by Klebsiella pneumoniae (anti-hsp60, anti-hsp65 and anti-hsp70), Pseudomonas aeruginosa (anti-hsp60, anti-hsp70) and Aspergillus fumigatus (anti-hsp65). We conclude that anti-hsp antibodies might be produced after SCT in relation to infection depending on etiological agents; however, transplant-related complications by themselves had a little impact.     

Successful hematopoietic stem cell transplantation in Farber disease

Farber disease, a lysosomal storage disorder, has a dismal prognosis leading to death with progressive granulomatous inflammation, even in patients without central nervous system involvement (type 2/3). We report the first successful hematopoietic stem cell transplantations in 2 patients with Farber disease type 2/3, resulting in resolution of symptoms.     

Long-term follow-up of IPEX syndrome patients after different therapeutic strategies: An international multicenter retrospective study

1. Download high-res image (308KB)
2. Download full-size image

     

Predicting an HLA-DPB1 expression marker based on standard DPB1 genotyping: Linkage analysis of over 32,000 samples

The risk of acute graft-versus-host disease (GvHD) after hematopoietic stem cell transplantation is increased with donor-recipient HLA-DPB1 allele mismatching. The single-nucleotide polymorphism (SNP) rs9277534 within the 3′ untranslated region (UTR) correlates with HLA-DPB1 allotype expression and serves as a marker for permissive HLA-DPB1 mismatches. Since rs9277534 is not routinely typed, we analyzed 32,681 samples of mostly European ancestry to investigate if the rs9277534 allele can be reliably imputed from standard DPB1 genotyping. We confirmed the previously-defined linkages between rs9277534 and 18 DPB1 alleles and established additional linkages for 46 DPB1 alleles. Based on these linkages, the rs9277534 allele could be predicted for 99.6% of the samples based on DPB1 genotypes (99.99% concordance). We demonstrate that 100% prediction accuracy could be achieved if the prediction utilized exon 3 sequence information. DPB1 genotyping based on exon 2 data alone allows no unambiguous rs9277534 allele prediction but was estimated to maintain 99% accuracy for samples of European descent. We conclude that DPB1 genotyping is sufficient to infer the DPB1 expression marker rs9277534 with high accuracy. This information could be used to select donors with permissive HLA-DPB1 mismatches without directly screening for rs9277534.