CD5-positive B cells after T cell depleted bone marrow transplantation.

CD5 is an antigen normally found on T cells and on a minority subpopulation of B cells in fetal spleen and tonsil and on the majority of cells in B-chronic lymphocytic leukaemia. Recent reports described the occurrence of large numbers of CD5-positive B cells in the peripheral blood after bone marrow transplantation (BMT). We examined the peripheral blood for CD5-positive B cells in 21 patients who underwent allogeneic BMT for leukaemia with marrow first depleted of T cells using anti-T monoclonal antibodies and complement mediated lysis.

Synthesis and regulation of the two human complement C4 genes in stable transfected mouse fibroblasts.

The major histocompatibility complex-linked human complement C4 genes are highly homologous in primary structure but give rise to products which differ in complement-activating function. In order to examine the synthesis, function, and regulation of these two genes independently, cloned C4A and C4B genes were transfected into mouse fibroblast L-cells. In the stable transfected cell lines, C4A and C4B are synthesized, undergo a complex series of post-translational modifications, and each functions appropriately in activation of the classical complement pathway.

B cell development and regulation after T cell-depleted marrow transplantation.

Antibody secreting B lymphocytes from immunized donors can be adoptively transferred after T cell-depleted marrow transplantation to produce protective levels of antibody in the recipient. We have investigated whether these transferred lymphocytes remain subject to continued clonal selection and subsequently became memory B cells even in the initial absence of T cells. Twenty-eight donor/recipient pairs were randomized pretransplant to be immunized or not with tetanus toxoid (TT).

Bone-marrow transplantation in high-risk acute lymphoblastic leukaemia in first and second remission.

Bone-marrow transplantation has been used in patients with acute lymphoblastic leukaemia (ALL) thought to be at high risk of relapse if managed with chemotherapy. Data from 444 ALL patients with one or more high-risk features at diagnosis were analysed to evaluate outcome after HLA-identical bone-marrow transplantation during first or during second remission. The 4-year actuarial probability of leukaemia-free survival was 45% (95% confidence interval 36-54%) for transplants in first remission compared with 22% (15-29%) for those in second remission (p less than 0.

Immune donors can protect marrow-transplant recipients from severe cytomegalovirus infections.

To study the importance of transferred immunity against cytomegalovirus (CMV) in allogeneic, HLA-matched, T-cell-depleted bone-marrow transplantation, the incidence, severity, and outcome of CMV infections were studied in 40 CMV-seropositive recipients in relation to the donors' immunity against CMV. There was no significant difference in the incidence of CMV infections between recipients of seropositive (n = 27) and seronegative (n = 13) marrow. However, the incidence of CMV pneumonitis (8/13 compared with 4/27; p less than 0.

Haemorrhagic cystitis in bone marrow transplantation patients: possible increased risk associated with prior busulphan therapy.

Following bone marrow transplantation employing conditioning including 'high-dose' cyclophosphamide, 65 patients were studied for the subsequent development of symptomatic haemorrhagic cystitis. There was no protection from the urothelial toxicity of cyclophosphamide metabolites afforded by the concurrent administration of 2-mercaptoethane sodium sulphonate (mesna) if timing errors in administration were made. Other factors which might increase the risk of haemorrhagic cystitis due to cyclophosphamide administration include the prior administration of busulphan to patients with chronic granulocytic leukaemia, in whom the incidence of haemorrhagic cystitis was 36% compared with 4% in all other patients.

Cytotoxic effects of tumour necrosis factor and gamma-interferon on acute myeloid leukaemia blasts.

We have studied the cytotoxic effects of recombinant tumour necrosis factor and recombinant gamma interferon on primary cultures of leukaemia cells. The agents were added alone or in a combination to cells from 17 patients. Eleven had acute myeloblastic leukaemia (6 at presentation, 5 at relapse), 4 had acute lymphoblastic leukaemia, one had hairy cell leukaemia, and 2 had chronic myeloid leukaemia--one of whom was in myeloid blast transformation.

Human large granular lymphocytes induce immunoglobulin synthesis after bone marrow transplantation.

Following T cell-depleted bone marrow transplantation, helper T cell numbers remain depressed for some months. Nonetheless, functional B cells can be adoptively transferred to the recipients of such grafts, where they continue to secrete antibody. We now show that immunoglobulin production by these transferred B cells is induced by activated large granular lymphocytes (LGL) which circulate in the recipients in substantial numbers during the immediate post-transplant period.

Mitozantrone-induced toxicity and DNA strand breaks in leukaemic cells.

By applying the fluorometric analysis of DNA unwinding (FADU) the in vitro effect of mitozantrone on DNA strand breaks was studied in seven different human leukaemic/lymphoma cell lines and in fresh leukaemic samples from seven patients with acute myeloid leukaemia refractory to conventional treatment. Pulse exposure to mitozantrone for 30 min invariably caused strand breaks. In the cell lines JM 1, KM3 and RPM I 8420 DNA strand breakage was progressive upon further incubation in drug free medium.

Pulmonary cryptosporidiosis occurring in a bone marrow transplant patient.

We describe what is believed to be the first bone marrow transplant patient, a 32-year-old man, in whom pulmonary cryptosporidiosis was associated with terminal respiratory failure. The diagnosis, treatment and postmortem histology are discussed together with a brief review of the literature.

Bone marrow transplantation in acute or chronic leukaemia.

Using T-depleted BM from HLA-identical sibling donors we have performed 36 BMTs since 5/83 in first remission (CR1) of acute leukaemia (AL). Standard conditioning for BMT consisted of Cy 60 mg/kg X 2 and 7.5 Gy single fraction TBI (n = 27).

The role of mitozantrone in the treatment of acute leukaemia.

Mitozantrone (MTZ; Novantrone), 1,4-dihydroxy-5,8-bis-[(2-[(2-hydroxyethyl)amino]ethyl]amino)-9,10- anthracenedione dihydrochloride (NSC 302739), is a synthetic anthracenedione with intercalating properties. Activity has been shown in preclinical studies in mice bearing intraperitoneal P388 and L1210 leukaemias and a variety of solid tumours. Three sequential studies to examine the activity, toxicity and scheduling of MTZ in acute leukaemia or chronic granulocytic leukaemia in myeloid transformation were undertaken in relapsed and/or refractory disease in patients up to the age of 70.

Nerve growth factor activates Thy-1 and neurofilament gene transcription in rat PC12 cells.

The effect of nerve growth factor (NGF) on the expression of neurofilament and Thy-1 genes in rat PC12 pheochromocytoma cells was examined at both the transcriptional and post-transcriptional levels. Addition of NGF to cultured PC12 cells produced increases in mRNAs corresponding to the 68 kd neurofilament protein (NF68) and the Thy-1 glycoprotein within 24 h, with maximal effects of some 90- and 45-fold stimulation (relative to beta-actin mRNA) being observed after 12 and 4 days of treatment, respectively. In addition, transcriptional run-off analyses using isolated nuclei showed that NGF treatment resulted directly in 8- and 4-fold increases in the rate of NF68 and Thy-1 gene transcription.

T-cell depletion of allogeneic bone marrow prevents acceleration of graft-versus-host disease induced by exogenous interleukin 2.

Highly purified human recombinant interleukin 2 (IL-2) markedly accelerated lethal GVHD in the H-2-identical B10.BR----CBA combination, but had no effect when the donor cells were depleted of mature (Thy-1.2-positive) T lymphocytes, indicating a strong immunopotentiating effect of IL-2 on mature T cells causing GVHD.

A515U: a prodrug of acyclovir with increased oral bioavailability.

The tolerance and pharmacokinetics of A515U, a xanthine oxidase-activated prodrug of acyclovir have been investigated in healthy volunteers and in two phase-I clinical studies in immunocompromised patients. In all cases the bioavailability of acyclovir following oral administration of A515U was substantially increased over that achieved in the same subjects with oral acyclovir itself. Plasma acyclovir levels were similar to those previously attainable only with intravenous acyclovir.

Standardization of T-cell depletion in HLA matched bone marrow transplantation.

The IBM 2991 Blood Cell Processor has been used to isolate a mononuclear cell (MNC) fraction from the marrow of 31 allogeneic donors. The MNC fraction was then incubated with a combination of two murine monoclonal antibodies MBG6 (CD6) and RFT8 (CD8) followed by two rounds of treatment with rabbit complement resulting in a marrow inoculum significantly reduced in the number of T-lymphocytes. We report here new specifications for the use of Ficoll-Metrizoate, the method used to calculate T-lymphocyte depletion and the details of our attempts to improve T-depletion.

Recovery of immunoglobulin isotypes following T-cell depleted allogeneic bone marrow transplantation.

After conventional bone marrow transplantation serum IgG, IgM and IgA levels fall from pre-transplant levels and may not return to normal for 3-12 months. In contrast IgE may rise to supranormal levels, an event that may be associated with graft-versus-host disease. We have investigated the recovery of immunoglobulin isotypes in the recipients of allogeneic marrows depleted of T-cells to prevent graft-versus-host disease.

The contribution of large granular lymphocytes to B cell activation and differentiation after T-cell-depleted allogeneic bone marrow transplantation.

Immunoglobulin and specific antibody levels are well maintained in the recipients of T-cell-depleted allogeneic bone marrow transplants (BMT), even though up to 99% of mature T cells are removed from the donor graft. For 3-8 weeks after the procedure, natural killer (NK) cells with an activated pattern of target cell killing have been shown to circulate in the recipient. This study investigates whether these recipient NK cells spontaneously secrete lymphokines that modulate B cell function in a way analogous to that of in-vitro-activated NK cells from normal individuals.

Production of anti-cytomegalovirus antibody following T-cell depleted bone marrow transplant.

The serological status to cytomegalovirus (CMV) was examined in 24 patients with no detectable CMV excretion, following T-lymphocyte depleted bone marrow transplantation. The results show that seropositive recipients continue to produce CMV antibody regardless of the serological status of the donor but that seronegative recipients fail to produce CMV antibody even when the donor is seropositive. These findings suggest that when an effective CMV vaccine becomes available vaccination of the donor would be unlikely to confer protection on the recipient but that vaccination of the recipient may achieve this.

Haemolysis after T-cell depleted bone marrow transplantation involving minor AB0 incompatibility.

9 recipients of T-cell depleted allogeneic bone marrows (8 group A, 1 group AB) from group 0 donors were monitored after transplantation. Free anti-A/B was demonstrable in 8 of the 9 recipients 10-19 d post-transplant, 5 patients developed a positive direct anti-globulin test and 7 showed a rise in bilirubin. The presence of antibody was generally unrelated to the infusion of incompatible plasma, although 2 patients who also received anti-CMV immunoglobulin subsequently shown to contain high titre IgG anti-A/B were more severely affected, sustaining a fall in Hb of up to 2 g/d.

Graft rejection following HLA matched T-lymphocyte depleted bone marrow transplantation.

Bone marrow graft rejection following HLA-matched bone marrow transplantation (BMT) for leukaemia has been a rare problem. However, with the introduction of T-lymphocyte depleted BMT, graft rejection is recognized as a new complication. At the Royal Free Hospital (RFH) in London T-depletion is achieved using two monoclonal antibodies with complement mediated lysis.

Salivary amylase and pancreatic enzymes in serum after total body irradiation.

Amylase increases in serum after total body irradiation (TBI). To investigate whether this increase is ascribable to pancreatic or salivary amylase, or both, we measured the post-irradiation activity concentration of the three major pancreatic enzymes (trypsin, amylase, and lipase) and of salivary-type amylase in serum from six leukemic patients given TBI before bone-marrow transplantation. Salivary amylase increased by as much as 50-fold after TBI, peaking on the first day after the irradiation.