Bernard Pollara

Adenosine-deaminase deficiency in two patients with severely impaired cellular immunity.

Abstract Two young unrelated girls with similar but not identical manifestations of immunological deficiency were found to have no measurable adenosine-deaminase (A.D.A.) enzyme activity in their red blood-cells. The red-cell A.D.A. levels in the parents of one child were about half normal and in the other set of parents about two-thirds normal, suggesting they may be heterozygous, and their affected children homozygous, for a mutant A.D.A. gene. Since the A.D.A. produced by normal lymphocytes is mainly the kind found in red cells, a causal association is postulated between the absence of A.D.A. and impaired lymphocyte function in certain patients with inherited immune disease. An alternative possibility of partial chromosomal deletion is also considered.

Inhibition of experimental autoimmune renal tubulointerstitial disease in guinea pigs by depletion of complement with cobra venom factor.

Passive transfer experiments have shown that autoimmune renal tubulointerstitial disease (RTD) in guinea pigs is caused by autoantibodies specific for tubular basement membrane (TBM). Further studies on the pathogenesis of this renal disease have implicated complement (C) as a possible mediator of antibody-dependent tissue injury because C3 is present along with IgG on the TBM. However, as shown in experiments with C4 deficient guinea pigs, no essential pathogenetic role could be ascribed to C4, suggesting the involvement of alternate pathway of complement activation. To delineate the role of C in this model, attempts were made to induce RTD by passive transfer in guinea pigs depleted of 95% of C with cobra venom factor (CoF). RTD could not be induced by anti-TBM antibodies alone. IgG and C3 must both be bound to TBM for a sufficient period of time before tissue injury can occur. An essential role for C participation in the pathogenesis of this model of autoimmune tubular nephritis is evident. Mononuclear cells, prominent in these lesions, appear to accumulate and proliferate in the renal cortex only when sufficient amounts of C are available. CoF did not seem to have a direct inhibitory effect on mononucular cells in this model.

717 MARROW TRANSPLANTATION (MTP) IN WISKOTT-ALDRICH SYNDROME (WAS): T CELL ENGRAFTMENT WITH CYCLOPHOSPHAMIDE (CY), COMPLETE ENGRAFTMENT WITH TOTAL BODY IRRADIATION

We studied two patients with WAS who had MTP with marrow from HLA-identical siblings. The first patient received 200 mg/kg CY over 4 days (Bach et al., Lancet 1:1364, 1968). We have followed this patient for 7 years. He has petechiae, bleeding in joints, and severe thrombocytopenia. IgM has remained low; antibody production to some antigens, lymphocyte response to mitogens and in vitro B cell functions are defective. Growth and development have been normal. The second patient received cytosine arabinoside 5 mg/kg/day ×7, 6-thioquanine 4 mg/kg/day ×7, and CY 50 mg/kg/day ×4 prior to MTP. In both patients after MTP, megakaryocytes, red cells, neutrophils, macrophages, and most B cells remained of recipient type while most T cells were of donor type. The second patient was retransplanted after preparation with anti-lymphocyte globulin, 2 ml IV daily x4, Procarbazine, 12.5 mg/kg/day × 3, and 800 R total body irradiation. Ninety days after the 2nd transplant, all nucleated marrow and blood cells, including B and T cells were of donor type; WBC 12,500, platelets 132,000/mm3, hemoglobin 9.8 gms % and reticulocytes 3.5%. No graft-versus-host disease was observed. We conclude that in marrow transplantation for WAS, 200 mg/kg CY may be inadequate preparation. In WAS, T cells may more readily be engrafted than other blood cells. Supported by USPHS GCRC #MO-1-R00749-05.

A Rapid Method for the Separation of Guinea Pig IgG1 and IgG2

Immunoelectrophoretically pure IgG1 and IgG2 were isolated from sera of guinea pigs with experimental autoimmune renal tubular disease by chromatography on QAE-Sephadex A-50 (Pharmacia Fine Chemicals, Piscataway, N.J. using a two-step elution. As judged by indirect immunofluorescence tests of the fractions, 70% of the original autoantibody activity to tubular basement membrane was recovered.

Experimental Autoimmune Renal Tubulointerstitial Disease

It is well established that many types of glomerular diseases are caused by immune complexes or by autoantibodies to glomerular basement membrane (GBM) (Wilson, this volume). In experimental models and in cases of human glomerulonephritis, there also has been immunopathologic evidence for deposition of immune reactants along the tubular basement membrane (TBM) (Andres et al., 1978) although their role in the pathogenesis of tubular lesions remained largely unrecognized until about 7 years ago. As in glomerular diseases, compelling evidence for immune mechanisms in renal tubular diseases in man was obtained only after convincing animal models had been established (Andres and McCluskey, 1975) (Table 1).

547 RETRANSFER (RT) IS A SAFE AND COST-EFFECTIVE MEANS OF IMPROVING NEONATAL INTENSIVE CARE UNIT (NICU) UTILIZATION

During an 18 mo. period, 1255 neonates were admitted to the only Level III (LIII) NICU in a 50,000 sq. mi. service area with 26,000 live births/yr and having 33 Level I (LI) and 5 Level II (LII) referring hospitals. 739 of 1255 patients were transported from referring hospitals. 524 were neonatal transports (NT) and 215 maternal transports (MT). 641 of 739 survived and were eligible for RT to referring hospitals after stabilization. 225/641 (35%) underwent RT; only 4 required NICU readmission. Duration of hospitalization was greater for infants < 2000 gms. birthweight (BW). Analysis of transport and RT patterns is shown below.Rates of RT to LI or LII were all < 50%; when used, RT safely decreased duration of NICU hospitalization.Conclusions: at an average LIII cost of

TISSUE ADENOSINE DEAMINASE ACTIVITY IN AN ADENOSINE DEFICIENT-COMBINED IMMUNODEFICIENCY DISEASE PATIENT

800/day and an LI/LII cost of

Auditory Function in Rat Pups Exposed Perinatally to Zidovudine (ZDV)|[dagger]| 693

400/day, RT represents

Characterization of the residual adenosine deaminating activity in the spleen of a patient with combined immunodeficiency disease and adenosine deaminase deficiency.

770,000 in potential annual savings. As shown here, analysis of regional referral patterns and use of RT aids optimal use of limited NICU resources.

Experimental Autoimmune Renal Cortical Tubulointerstitial Disease in Guinea Pigs Lacking the Fourth Component of Complement (C4)

Patients with combined immunodeficiency (CID) and adenosine deaminase (ADA) deficiency may have low or absent ADA activity in various tissues. Hirschhorn et al. have shown that the activity in patient fibroblasts is a mutant form of ADA (PNAS 73: 213, 1976). Others have suggested that the deficiency reflects an inhibition of ADA (Trotta et al., PNAS 73: 104, 1976). We have isolated and in part characterized properties of the residual ADA from spleen of one patient. Radioimmunoassay for ADA protein in erythrocytes and spleen extracts of the patient showed no cross-reacting protein. Chromatography of ADA-CID spleen extracts produced a fraction with adenosine deaminating activity which could not be adsorbed on affinity or anti-ADA columns. The deaminating activity could not be inhibited with erythro-9-(2-hydroxyl-3-nonyl) adenosine. The pH optimum activity curve and Km differed from normal spleen ADA. A similar fraction, representing about 1% of the total ADA activity, was isolated from normal spleen extracts. We conclude that the adenosine deaminating activity observed in patients with ADA-CID is not ADA. The activity may be due to an enzyme whose principal substrate may be another metabolite. The presence of this non-ADA deaminating activity in normal spleen suggests that the observed activity in ADA-CID tissue is not due to a mutant form of ADA and that the inhibited enzyme activity reported by Trotta et al. may be due to an amplification of this non-ADA deaminating enzyme.