EBV‐directed viral‐specific T‐lymphocyte therapy for the treatment of EBV‐driven lymphoma in two patients with primary immunodeficiency and DNA repair defects

Children with ataxia telangiectasia (AT), a primary immunodeficiency caused by mutations in ATM, which is critical for repairing DNA defects, are at risk for the development of hematologic malignancy, frequently driven by infection with Epstein‐Barr virus (EBV). Conventional chemotherapy is poorly tolerated by patients with AT, with excessive toxicity even when doses are reduced. Here, we report on two patients with AT and EBV‐positive neoplasms who were treated with EBV‐targeted viral‐specific T cells (VST). One patient had a prolonged complete response to VSTs while the other had a partial response. Therapy was well tolerated without infusion toxicity or graft‐versus‐host disease.     

Hyponatremia caused by thiazide diuretics: be aware of drug combinations which enhance this effect

In 3 patients, two women aged 88 and 82 and a man aged 76, the consciousness became disrupted due to a severe hyponatraemia, after a thiazide diuretic had been combined with another drug without laboratory control. After a change in medication, the laboratory values and the patients' conditions normalised. Severe hyponatraemia is a well known but rare complication of thiazide therapy. It has a significant mortality and morbidity rate. The risk is greater for elderly women. This effect on serum sodium can be enhanced by the use of other drugs like furosemide, carbamazepine, paroxetine and NSAIDs. That a patient uses a thiazide is sometimes overlooked when a combined preparation of other drugs is prescribed. Diuretic serum electrolytes should be monitored once treatment with thiazide has been started, especially in elderly patients taking other drugs.     

Influence of combinations of cytokines on proliferation of isolated single cell-sorted human bone marrow hematopoietic progenitor cells in the absence and presence of serum.

Human mast cell growth factor (MGF, a c-kit ligand) and colony stimulating factors (Epo, GM-CSF, G-CSF, IL-3) were assessed in the absence or presence of serum for stimulation in semi-solid medium of single CD34 , CD34 HLA-DR+, or CD34 HLA-DR+CD33- cells sorted per microtiter well. The % of wells containing CFU-GM and erythroid containing (BFU-E and CFU-GEMM) colonies increased in proportion to the number of cytokines added. In the presence of serum, 1, to 4 cytokine combinations resulted in respective increases in cloning efficiencies of 10 to 21.0, 19.5 to 31.5, 35.8 to 42.9, and 46.3 to 60.0%. MGF had little effect by itself, but did act in combination with CSFs to enhance numbers and size of the colonies from isolated single cells. High cloning efficiencies were also obtained in the absence of serum when multiple cytokines were used. The results demonstrate that MGF and CSFs can act directly on the proliferation of single hematopoietic progenitor cells in the absence of accessory cells and serum.     

The small GTPase Rac is involved in clustering of hippocampal neurons and fasciculation of their neurites

In hippocampal neurons cultured from brains of newborn rats, the glutamate receptor agonist N-methyl-d-aspartate induced the clustering of neuronal perikarya and the fasciculation of neurites. In addition, N-methyl-d-aspartate activated the small GTPase Rac1. Other stimuli of Rac activity, such as the Rho kinase inhibitors Y-27632, H-1152, and H89, as well as the cytotoxic necrotizing factor-1 from Escherichia coli, also caused neuronal clustering and neurite bundling. In neurons transiently transfected with dominant negative Rac1N17 neither N-methyl-d-aspartate nor Y-27632 induced clustering and fasciculation. In addition, the PI3-kinase inhibitors wortmannin and LY-294002 prevented these effects, as did a dominant negative form of p110PI3-K. Time-lapse microscopy showed that lethal toxin from Clostridium sordellii, which inhibits Rac, and wortmannin blocked the neuronal migration induced by Y-27632. In contrast, only lethal toxin reversed the clustering and fasciculation induced by pre-treatment with Y-27632. This effect of the toxin may be due to inactivation of Ras, since FTI-277, which prevents the farnesylation of Ras and thereby inactivates the GTPase, also dissolved the preformed clusters. We suggest that active Rac and a PI3-kinase synergistically induce neuronal migration, whereas a Ras isoform is responsible for the lasting attachment of neurons necessary for clustering and neurite fasciculation.     

Mineralocorticoid excess due to precursors only in a patient with adrenal cortical carcinoma.

A patient with carcinoma of the adrenal cortex presented with mineralocorticoid excess due to hypersecretion of deoxycorticosterone, which is exceedingly rare. Backache was the only symptom and an unexplained hypokalaemia was the only sign. Because of the abnormal synthesis of steroid precursors in these tumours a urinary steroid profile may be helpful as a diagnostic tool in such cases.     

Sexual disturbances arising from multiple sclerosis

Multiple sclerosis patients under the age of 50 with only a slight handicap were very often found to have disturbances of sexual functions (among male patients 71.4%; among female patients 73.9%; control group 18.7%). In 19.6% of the cases, the disturbances were of a serious nature. These disturbances, as well as disturbances of function of bladder and colon, were found to be mainly brought about by separate damage in the lumbosacral region of the spinal cord. The incidence of these disturbances bore no relationship to the degree of motor disturbances, age of the patient or duration of the disease.     

Incidence of HIV infection in monocyte subpopulations characterized by CD4 and HLA-DR surface density.

OBJECTIVE: The purpose of this study was to determine any correlation between the expression of CD4 antigen on the surface of monocytes, and the frequency with which these cells are infected with HIV. DESIGN: CD4 surface expression on monocytes is significantly less than that expressed on CD4+ lymphocytes. Nevertheless, all monocytes express the HIV CD4 receptor and infected individuals have a significant decrease in the number of monocytes that express a higher density of CD4 surface fluorescence. METHODS: Three-color flow cytometric analysis was used to characterize monocyte-enriched peripheral blood mononuclear cells (PBMC) in terms of surface expression of CD4, CD14 (macrophage antigen), and class II major histocompatibility antigen (HLA-DR). HLA-DR+ monocytes from HIV-positive individuals were sorted into two subpopulations based on either 'bright' or 'dim' CD4 surface expression. A polymerase chain reaction (PCR) assay was used to detect the presence of proviral HIV sequences within the DNA from 10(5) cells from each sorted population. RESULTS: Post-sort analysis revealed that the dim CD4+ monocyte subset expressed dim HLA-DR surface antigen, while the bright CD4+ monocyte subset contained both bright and dim HLA-DR+ cells. PCR results showed that four out of eight dim CD4+ monocyte subsets contained proviral HIV DNA, compared with one out of eight bright CD4+ monocyte subsets.     

Transplantation of a fetus with paternal Thy-1(+)CD34(+)cells for chronic granulomatous disease

A fetus diagnosed with X-linked chronic granulomatous disease was transplanted with Thy-1(+)CD34(+) cells of paternal origin. The transplant was performed at 14 weeks gestation by ultrasound guided injection into the peritoneal cavity. The fetus was delivered at 38 weeks gestation after an otherwise uneventful pregnancy. Umbilical cord blood was collected and used to determine the level of peripheral blood chimerism as well as levels of functional engrafted cells. Flow cytometry was used to detect donor leukocytes identified as HLA-A2(-)B7(+) cells, whereas recipient cells were identified as HLA-A2(+)B7(-) cells. No evidence of donor cell engraftment above a level of 0.01% was found. PCR was used to detect HLA-DRB1*15(+) donor cells among the recipient's HLA-DRB1*15(-) cells, but no engraftment was seen with a sensitivity of 1:1000. The presence of functional, donor-derived neutrophils was assessed by flow cytometry using two different fluorescent dyes that measure reactive oxygen species generated by the phagocyte NADPH oxidase. No evidence of paternal-derived functional neutrophils above a level of 0.15% was observed. Peripheral blood and bone marrow samples were collected at 6 months of age. Neither sample showed engraftment by HLA typing using both flow cytometry and PCR. Functional phagocytes were also not observed. Furthermore, no indication of immunological tolerance specific for the donor cells was indicated by a mixed lymphocyte reaction assay performed at 6 months of age. While there appears to be no engraftment of the donor stem cells, the transplant caused no harm to the fetus and the child was healthy at 6 months of age. Analyses of fetal tissues, obtained from elective abortions, revealed that CD3(+) T cells and CD56(+)CD3(-) NK cells are present in the liver at 8 weeks gestation and in the blood by 9 weeks gestation. The presence of these lymphocytes may contribute to the lack of donor cell engraftment in the human fetus.