Outcome of T cell-depleted transplantation after conditioning with an intensified regimen in patients aged 50 years or more is comparable with that in younger patients

One hundred and thirty-one patients were transplanted for AML-CR1, ALL-CR1 or CML-CP1 after conditioning with 120 mg/kg body weight cyclophosphamide and 2 x 4.5 Gy TBI. Conditioning was intensified with the addition of 42 mg/m2 idarubicin. Grafts were T cell-depleted using counterflow centrifugation. Donors were HLA-identical siblings. We compared outcome of BMT in 109 patients aged less than 50 (median, 35) years with that of 22 patients with an age of 50 years or more (median, 53 years). For the patients aged <50 years, 2-year probabilities of treatment-related mortality, relapse, survival and leukemia-free survival were 26% (95% CI, 17% to 35%), 26% (95% CI, 17% to 35%), 64% (95% CI, 55% to 73%), and 56% (95% CI, 47% to 65%). For the patients aged > or =50 years, these figures were 13% (95% CI, 0% to 30%), 24% (95% CI, 6% to 42%), 66% (95% CI, 46% to 86%), and 67% (95% CI, 47% to 87%), respectively. Outcome did not differ significantly between the two age groups. TRM was within the range of that reported in the literature for recipients of T cell-depleted grafts. We conclude that T cell-depleted transplantation after a conditioning regimen that was intensified with the addition of idarubicin is feasible in patients aged > or =50 years. For this age group of patients, results of nonmyeloablative regimens should be compared with that obtained with T cell-depleted grafts.     

BCG-induced trained immunity in NK cells: Role for non-specific protection to infection

Adaptive features of innate immunity, also termed ‘trained immunity’, have recently been shown to characterize monocytes of BCG vaccinated healthy volunteers. Trained immunity leads to increased cytokine production in response to non-related pathogens via epigenetic reprogramming of monocytes. Recently, memory-like properties were also observed in NK cells during viral infections, but it is unknown if memory properties of NK cells contribute to trained immunity due to BCG vaccination.     

CD4 deficiency in myelodysplastic syndrome with monosomy 7

Abstract We describe a patient with myelodysplastic syndrome with monosomy 7 presenting with a T-cell defect. He suffered from infections from the age of 10 years, when a CD4 deficiency and impaired lymphoproliferative responses in vitro were found. The only symptom of a myelodysplastic syndrome at that time was thrombocytopenia with giant platelets. Monosomy 7 was found in the bone marrow cells. At the age of 11 years he developed other characteristics of monosomy 7 including splenomegaly and anaemia. Some months later leukaemia was diagnosed.Conclusion In non-HIV CD4 deficiency myelodysplastic syndrome has to be considered.     

Parenteral lipids modulate leukocyte phenotypes in whole blood, depending on their fatty acid composition

To characterize the immunological effects of various lipids that are applied as part of total parenteral nutrition (TPN) formulations, we analyzed phenotypical changes in leukocytes following lipid exposure. Importantly, the study was performed with whole blood in order to prevent the functional changes that are induced by isolation procedures. Briefly, blood samples from 10 healthy volunteers were incubated with lipids containing pure long-chain triglycerides (L), mixed long- and medium-chain triglycerides (LM), synthetic structured lipids (SL), or emulsions based on olive oil (OO), or fish oil (FO). After immune fluorescent staining, leukocyte phenotype characteristics were analyzed by flowcytometry. Exposure to LM increased in a dose-dependent manner the expression of membrane surface markers for adhesion (CD11b) and degranulation (CD66b), while decreasing CD62L, on neutrophils and monocytes. These findings demonstrate that LM activates leukocytes in peripheral whole blood. On the other hand, decreased expression of activation markers was observed with L and FO. Lipids effects on the phenotype of T lymphocytes and Natural Killer cells were not seen during incubation for up to 4 h. These results indicate that (i) the composition of TPN formulations with regard to lipid structure has implications for the function of exposed immune competent cells and (ii) medium-chain triglycerides, which have been regarded as functionally inert deliverers of fuel calories, have distinct biological effects.     

Analysis of 127 stem cell donations of the regional Bone Marrow Donor Bank Europdonor Nijmegen,The Netherlands

In December 2000, the Bone Marrow Donor Bank Europdonor Nijmegen in the Netherlands celebrated its tenth anniversary. We describe the organisation and activities in the first 10 years of this regional bone marrow donor bank. A concise inquiry was sent to all transplant centres who had received a graft from our donors. Response rate was 88% and data were available from 127 recipients. Three donors donated twice to different patients. Median age of the 124 donors (42 females and 82 males) was 37 years and 30 years for the 127 recipients (48 females and 79 males). Time interval between first request of a blood sample and collection of bone marrow varied from 13 to 695 days (median, 113 days). All but two donors received general anaesthesia for 25-120 min (median; 60 min). Hospital stay has been reduced to 24 h. Most donors experienced pain from the collection sites for 3-5 days. However, 9 donors (7%) suffered from pain for 2-3 weeks. All but two donors (98%) were willing to donate a second time for the same patient and 119 (96%) donors wished to remain in the register. The number of nucleated cells (NC) in the collected marrow varied from 0.2 to 8.3 ×10 8 /kg body weight of the recipient (median, 3.5 ×10 8 /kg ) with 6.4-470.0 ×10 4 CFU-GM/kg body weight of the recipient (median, 18.0 ×10 4 /kg body weight). The 3-year projected probability of survival of the 127 recipients transplanted with marrow from donors provided by Bone Marrow Donor Bank Europdonor Nijmegen was 27 ±9% ( ±95% CI).     

Common variable immunodeficiency (CVID) in a family: an autosomal dominant mode of inheritance

BACKGROUND: Common variable immunodeficiency (CVID) is characterised by a late onset deficiency of immunoglobulins resulting in recurrent infectious and non-infectious ailments. Most cases are sporadic but occasional familial clustering has been described. We present an extensively affected family with CVID in three consecutive generations. METHODS: We conducted a study in this family to establish clinical phenotype, to clarify the mode of inheritance and to attempt to characterise the immune disturbance by determining immunoglobulin concentrations and B- and T-cell analysis. RESULTS: We describe six patients with CVID in three consecutive generations. In addition, we encountered 10 family members with dysimmunoglobulinemia. B-cell counts were normal, but T-cell analysis showed slightly abnormal results. CONCLUSIONS: The six cases of overt late onset hypogammaglobulinemia are compatible with an autosomal dominant mode of inheritance. The family members with dysimmunoglobulinemia may be at risk to develop overt CVID in the future, in view of the gradual course of progression of the disease in the clinically affected family members. B- and T-cell analysis are inconclusive though may support a possible defect in T-cell function to be involved. To further study this remarkable family and attempt to clarify pathogenesis, we are planning DNA linkage analysis in the near future.     

Perioperative replacement therapy in haemophilia B: An appeal to “B” more precise

Introduction

Haemophilia B is caused by a deficiency of coagulation factor IX (FIX) and characterized by bleeding in muscles and joints. In the perioperative setting, patients are treated with FIX replacement therapy to secure haemostasis. Targeting of specified FIX levels is challenging and requires frequent monitoring and adjustment of therapy.

Aim

To evaluate perioperative management in haemophilia B, including monitoring of FIX infusions and observed FIX levels, whereby predictors of low and high FIX levels were assessed.

Methods

In this international multicentre study, haemophilia B patients with FIX < 0.05 IU mL^?1 undergoing elective, minor or major surgical procedures between 2000 and 2015 were included. Data were collected on patient, surgical and treatment characteristics. Observed FIX levels were compared to target levels as recommended by guidelines.

Results

A total of 255 surgical procedures were performed in 118 patients (median age 40 years, median body weight 79 kg). Sixty percent of FIX levels within 24 hours of surgery were below target with a median difference of 0.22 IU mL^?1 [IQR 0.12‐0.36]; while >6 days after surgery, 59% of FIX levels were above target with a median difference of 0.19 IU mL^?1 [IQR 0.10‐0.39]. Clinically relevant bleeding complications (necessity of a second surgical intervention or red blood cell transfusion) occurred in 7 procedures (2.7%).

Conclusion

This study demonstrates that targeting of FIX levels in the perioperative setting is complex and suboptimal, but although this bleeding is minimal. Alternative dosing strategies taking patient and surgical characteristics as well as pharmacokinetic principles into account may help to optimize and individualize treatment.

     

Relationship between internalization and cytotoxicity of ricin A-chain immunotoxins

Immunotoxins (ITs) appear to vary considerably in their killing efficiency towards antigen positive cells. In order to unravel the mechanisms underlying these differences, the parameters responsible for these variations were studied. The efficacy of the monoclonal antibodies (MoAbs) WT32 (CD3), OKT4 (CD4), T101 (CD5), WT1 (CD7) and WT82 (CD8) conjugated to ricin A-chain was expressed by the extent of protein synthesis inhibition of four leukaemic T cell lines (CEM, GH1, Jurkat and HPB-ALL). Large differences in cytotoxicity were observed. Efficient inhibition of protein synthesis was seen with anti-CD3 IT, anti-CD5 IT and anti-CD7 IT. In these cases the cytotoxicity was related to the antigen density on the target cell membrane. Anti-CD4 IT inhibited poorly and anti-CD8 IT was ineffective, even on cell lines with a high expression of the corresponding antigen. When antigen density and cytotoxicity were plotted for all CD antigens, no correlation could be found. Subsequently, internalization was studied with 125Iodine-labelled antibodies. Anti-CD7 showed the fastest internalization rate, followed by anti-CD3 and anti-CD5. Anti-CD4 and anti-CD8 antibodies were respectively moderately and hardly internalized. When the absolute amount of internalized MoAb was calculated, a highly significant correlation with cytotoxicity was found. We conclude that the degree of antigen expression is not so important as the absolute amount of antibody internalized in predicting the efficacy of ITs.