Eosinophilic folliculitis: an important differential diagnosis after allogeneic bone-marrow transplant.

Eosinophilic folliculitis (EF) is a descriptive histopathological term applied to a heterogeneous group of disorders. In EF, the characteristic histopathological features are eosinophilic spongiosis and pustulosis involving the infundibular region of the hair follicle. EF may be seen in association with bacterial and fungal infection, drug reactions and haematological disorders. However, in those conditions, the histopathological changes are rarely restricted to the infundibula but in most cases include a moderate to dense perifollicular or even diffuse dermal infiltrate of lymphocytes, or eosinophilic or neutrophilic granulocytes. We present two cases of EF after mini-allogeneic bone-marrow transplantation (BMT) in order to highlight this rare and perhaps under-recognized clinical association.

Primary acute myeloid leukaemia blasts resistant to cytokine-induced differentiation to dendritic-like leukaemia cells can be forced to differentiate by the addition of bryostatin-1.

Primary acute myeloid leukaemia (AML) blasts can be induced to differentiate into dendritic-like leukaemia cells (DLLC) by culture with certain cytokine combinations. DLLC offer potential for use as autologous vaccines based on their ability to present putative leukaemia-specific antigens to T cells. It has been reported, however, that in around 30-50% of AML cases the leukaemia cells are not capable of undergoing DLLC differentiation. The purpose of this study was to identify the features that represent successful DLLC differentiation and, for those cases shown to be resistant to cytokine-induced differentiation, to use differentiating agents in an attempt to overcome the differentiation block. Leukaemia cells derived from 42 patients with AML were cultured in vitro with cytokines GM-CSF, IL-4 and TNFalpha/CD40L. In 22 cases the leukaemic cells underwent DLLC differentiation based on characteristic morphological changes and expression of costimulatory and dendritic cell-associated molecules. Four cases were not evaluable because of poor viability over the culture period. The remaining 16 cases failed to show evidence of DLLC differentiation. Many of these differentiation resistant cases were associated with poor risk karyotypic features. Nine of the resistant cases were selected for further study. Differentiating agents trichostatin (TSA), azacytidine (AZA) and bryostatin (BRYO) were used in combination with cytokines for the first 96 h of the culture period. Bryostatin (BRYO) alone was shown to be capable of overcoming differentiation resistance and allowing DLLC differentiation to proceed.

Recombinant coagulation factors.

The advent of technology that has allowed production of recombinant proteins on an industrial scale has revolutionized haemophilia care. Recombinant coagulation factors, as opposed to their plasma-derived counterparts, have a very low risk for transmission of infectious agents and their use should eradicate the threat of infection from viruses such as hepatitis C and the human immunodeficiency virus. This review outlines the manufacturing process involved in the production of the recombinant coagulation factors and the trials to date that have been performed to establish their safety and efficacy. We also discuss some of the issues involved in the change to use of recombinant coagulation factors, such as viral safety and potential immunogenicity.

Leucocyte depletion of blood components.

Universal leucocyte depletion has been implemented in the UK and several other European countries as a precautionary measure against the potential risk of transmission of variant Creutzfeldt-Jakob disease by blood transfusion. Leucocyte depletion had previously only been recommended for a relatively small proportion of transfusion recipients based on clinical and experimental evidence showing clinical benefit. However there is now increasing evidence to support its value in preventing transfusion transmission of infectious agents and in reducing some of the adverse immunomodulatory effects of allogeneic transfusion. The financial costs of providing universal leucocyte depletion are substantial, but, if it transpires that leucocyte depletion has a beneficial effect in reducing, for example, postoperative infection rates, then the health economic gains in this patient group alone may largely or wholly offset these financial costs. The experience in the UK and other European countries in terms of these collateral clinical benefits will help other countries, where the risk of variant Creutzfeldt-Jakob disease may not be so great, to decide whether to similarly adopt universal leucocyte depletion.

Lactobacillus rhamnosus infection in a child following bone marrow transplant.

We report a case where Lactobacillus rhamnosus was isolated from pericardial effusion and blood in a child following a bone marrow transplant for aplastic anaemia. A resume of cases in which this organism has been implicated as a pathogen is also presented.

Fusion hybrids of dendritic cells and autologous myeloid blasts as a potential cellular vaccine for acute myeloid leukaemia.

We assessed the potential of tumour cell/dendritic cell fusion hybrids to generate in vitro anti-leukaemic T-cell responses following co-culture with autologous remission lymphocytes in six patients with acute myeloid leukaemia (AML). Comparison was made to anti-leukaemic responses induced by mature dendritic cells (mDC) co-cultured with autologous, irradiated myeloid blasts. Fusion hybrids induced anti-leukaemic T-cell immune responses in three of six patients. Tumour-pulsed mDC induced T-cellular responses in two other patients. Only one of six patients remission lymphocytes failed to develop leukaemia-directed immune responses following stimulation with either construct. Anti-proliferative properties of fusion hybrids against allogeneic lymphocytes were observed in mixed lymphocyte-leukaemia reactions and were found not to be specific to the cell fusion partners and did not prevent the ability of AML-mDC heterokaryons to induce autologous anti-leukaemic cytotoxicity. We conclude that tumour cell/dendritic cell fusion hybrids hold promise as a cellular vaccine for AML.

Cellular therapy for haematological malignancies.

The aim of this review was to summarize the recent progress made in the field of cellular therapeutics in haematological malignancy. The review also examined the role that the National Transfusion Services might play in the manufacture of new cellular therapeutic agents, given both their expertise in the safe provision of blood products and their possession of accredited cell manipulation facilities. Cellular therapy is entering an era in which novel cellular products will find increasing clinical use, particularly in the areas of haematopoietic stem cell transplantation and immunotherapy. The production of novel cell-based therapies, both in Europe and North America, is now under strict regulatory control and therefore collaboration with the National Transfusion Services in the manufacture of these agents may well be beneficial if the production standards demanded by the regulatory authorities are to be fulfilled.

Gene transfer to primary acute myeloid leukaemia blasts and myeloid leukaemia cell lines.

The transfer of genes encoding co-stimulatory molecules and/or cytokines to leukaemia cells in order to create autologous tumour vaccines represents a potential immunotherapeutic strategy for treating acute myeloid leukaemia (AML). One of the essential requirements for this strategy if it is to be applicable in a clinical setting is a high efficiency of gene transfer to primary human AML blasts. Using green fluorescent protein (GFP) as a reporter gene, we have systematically evaluated a variety of physical, chemical and viral vector-based gene transfection systems in order to determine which gave the highest gene transfer efficiency to myeloid leukaemia cell lines and primary AML blasts. Transfection efficiency was low for all the physical and chemical transfection methods tested. Retroviral vector-based infection gave a high efficiency of gene transduction in two of the four leukaemia cell lines (KG1a and U937), but was low in primary AML blasts. An adenoviral vector gave a high transduction efficiency in all of the leukaemia cell lines with the exception of the HL60. In primary AML blasts, derived from 19 patients, gene transduction efficiency was variable, ranging from 1.1% to 67.1% (mean 12.1%). Following culture in cytokines GM-CSF/IL-4/CD40L, which induced differentiation of AML blasts to dendritic-like cells, transduction efficiency was increased between two- and eightfold in 6 out of the 15 cases that underwent differentiation.

Multiple myeloma in three siblings.

There have been 53 reported instances of myeloma occurring in more than one family member but in only three of these reports have three siblings been affected (Alexander & Benninghoff 1967; Maldonado & Kyle 1974; Horwitz et al. 1985; Crozes-Bony et al. 1995). We report a family where three siblings had an unequivocal diagnosis of myeloma made over a period of six years. The paraprotein isotype was IgG kappa in two of the siblings and kappa light chain only in the remaining sibling. The importance of these cases lies in the fact that they are highly suggestive of a genetic predisposition to the development of myeloma. Because of the high prevalence of p53 abnormalities in certain types of familial cancer screening for p53 mutation in exons 5-8 was performed by denaturing gradient gel electrophoresis (DGGE) on DNA extracted from the bone marrow of the three siblings. Although a p53 mutation was identified in one of the siblings it was felt to represent a somatic as opposed to a germline mutation.