Richard J. Pickering

Effects on the neonate of prednisone and azathioprine administered to the mother during pregnancy

We have reported a male infant whose mother was treated throughout pregnancy with large doses of azathioprine and prednisone for chronic renal disease. At birth, the child manifested lymphopenia, diminished thymic radiographic shadow, and low serum concentrations of immunoglobulins M and G. Cytomegalovirus (CMV) was cultured from maternal urine and from the infants urine at 10 weeks of age. The patient developed satisfactorily in the postnatal period and his immune status returned to normal. More than one year after birth the child appears to have developed adequately and, with the exception of low serum IgA concentration and persistent CMV infection, appears to have intact immunologic responses. Our observations in this patient and information derived from a literature review emphasize the need for short-and long-term clinical and laboratory evaluation of infants born to mothers who have been receiving immunosuppressive and potentially carcinogenic drugs during pregnancy.

Activation of the alternative complement pathway by enveloped viruses containing limited amounts of sialic acid

Abstract We have demonstrated that purified enveloped viruses grown in MDBK cells, such as influenza virus, Simian virus 5 (SV5), and vesicular stomatitis virus (VSV) grown in the presence of SV5 activate the alternative complement pathway, whereas VSV grown in BHK21-F or HKCC cells and Sindbis virus grown in BHK21-F cells do not. A direct correlation between the amount of sialic acid associated with the viral surface and its ability to activate the alternative complement pathway has been demonstrated. Our results indicate that enveloped viruses that lack sialic acid are efficient activators of the alternative complement pathway, whereas those with ≥10 μg of sialic acid/mg of protein do not. Enveloped viruses with 5–10 μg of sialic acid/mg of protein are intermediate in their ability to activate the alternative complement pathway. The results of our experiments employing different enveloped viruses with biologically derived sialic acid content support the hypothesis ( D. T. Fearon, 1978 , Proc. Nat. Acad. Sci. USA 75 1971–1975) that sialic acid is a key membrane constituent for modulating activation of C3 via the alternative complement pathway.

Genetic deficiency of the second component of complement (C2) associated with systemic lupus erythematosus: Relation of the complement abnormality and disease manifestations☆

Abstract Systemic lupus erythematosus (SLE) was documented in a patient with genetic deficiency of the second complement component (C2). A review of disease manifestations in this patient and in others with SLE and genetic C2 deficiency previously reported on suggest that many findings such as fever, skin lesions, central nervous system involvement, the presence of autoantibodies and leukopenia occur in patients with SLE independent of whether or not they have C2 deficiency. However, renal disease appears mild in patients with genetic C2 deficiency despite the presence of immunoglobulins and complement components demonstrated in glomeruli by immunofluorescent microscopy. Electron microscopic study of glomeruli from the patient we describe showed electron-dense deposits consistent with the immunofluorescent findings. Tubuloreticular inclusion bodies identical to those previously seen in patients with SLE were also observed. Analysis of serum complement components and tissue deposition of complement components suggests activation of both the classic and alternative complement pathways in patients with C2 deficiency. The clinical, pathologic and complement findings in this group of patients support the hypothesis that although SLE is similar in patients with and without C2 deficiency, renal disease remains mild when this classic pathway component is not present. Histocompatibility antigen analysis of the family of the propositus confirmed the association of C2 deficiency with an A10, B18 haplotype, and with a haplotype not previously associated with C2 deficiency, AW32, BW40. One sibling with this latter haplotype has normal C2 levels, presumably as a result of recombination.

Identification of the α-γ subunit of the eighth component of complement (C8) in a patient with systemic lupus erythematosus and absent C8 activity: Patient and family studies☆

Abstract The serum of a patient with systemic lupus erythematosus (SLE) and no known history of Neisseria infections lacked the activity of the eighth component of complement (C8). Antisera specific for human C8 identified in the serum of the patient and four relatives, whose C8 activity was well below normal, cross-reacting material which differed from normal C8 in electrophoretic mobility and molecular size. Electrophoresis of this C8-related material eluted from a F(ab′)2 anti-C8 immunoadsorbent column revealed the presence of the α-γ subunit of C8. That this material was intact α-γ subunit was shown by successful reconstitution of C8 hemolytic activity in the probands serum by purified β subunit. We conclude that the C8-related antigen in the serum of the proband and four apparently heterozygous relatives is the α-γ subunit of C8. The presence or absence of the α-γ subunit in C8-deficient individuals does not appear to influence the type of disease to which they are predisposed since we were unable to find similar C8 antigen in another C8-deficient patient with SLE. Similarly, susceptibility to Neisseria infections has been reported in individuals with and without functionless C8 antigen. Histocompatibility (HLA) studies showed that C8 deficiency in this family is not inherited in association with HLA antigens and the patient does not have the DR antigens commonly associated with SLE.

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.

C5a and antigen-induced tracheal contraction: Effect of a combination of an antihistamine and cyclo-oxygenase inhibitors

Our previous studies with C5a, a cleavage product of the fifth component of complement, have shown that the antihistamine diphenhydramine and the cyclo-oxygenase inhibitor aspirin do not inhibit the C5a-induced contraction of isolated guinea pig trachea (Regal, Eastman & Pickering, 1980; Regal & Pickering, 1981). We investigated the effect of cyclo-oxygenase inhibitors in the presence of diphenhydramine to determine if cyclo-oxygenase products were contributing to the contraction beyond any effect they might have on histamine release. A combination of a cyclo-oxygenase inhibitor and diphenhydramine caused a delay in onset and decrease in magnitude and duration of the C5a-induced contraction. Indomethacin itself also caused a slight inhibition. In contrast, a combination of aspirin and diphenhydramine did not inhibit the initial portion of antigen-induced tracheal contraction any more than diphenhydramine alone and enhanced the later portion just as aspirin alone. Cross tachyphylaxis experiments demonstrated that antigen pretreatment significantly inhibited a subsequent C5a-induced tracheal contraction, though C5a pretreatment did not affect a subsequent antigen-induced contraction. Thus, cyclo-oxygenase products do contribute to C5a-induced tracheal contraction, and histamine participation in the presence of cyclo-oxygenase inhibitors is suggested. Our studies demonstrate the dissimilarities of C5a and antigen-induced contraction as regards inhibition by aspirin plus diphenhydramine, yet suggest common pathways leading to the contractile response as evidenced by cross tachyphylaxis experiments.

Abnormalities of the complement system in Reye syndrome

Sixteen patients with Reye syndrome had diminished concentration of serum complement proteins and/or hemolytic activity in the earliest blood sample. All 12 studied with hemolytic methods had significantly reduced C1 activity; total hemolytic complement activity was reduced in only three. Low Cl activity was accompanied by equivalent reduction of Cls in 11 of 12 patients; Clq was less than normal in only two of 12. Decreased levels of at least one other classical pathway complement hemolytic activity or protein concentration were found in 13 patients, whereas factor B or the alternate complement pathway was normal or elevated in the ten patients studied. The consistent reduction of Cls protein concentration in Reye syndrome suggests that early metabolic abnormalities regularly affect the production or catabolism of this protein. Although normal serum Clq concentration in the majority of these patients does not support an immune pathogenesis, decreased Clq, C4, and C2 in three patients does suggest that immune mechanisms may be responsible for the serum complement abnormalities in this latter group of patients.

Enhancement of Granulocyte Oxidative Metabolism in Sera from Patients with C2 Deficiency and Systemic Lupus Erythematosus

Serum or plasma from 3 patients with C2 deficiency (C2D) and systemic lupus erythematosus (SLE) significantly enhanced chemiluminescence and superoxide anion production by polymorphonuclear leukocytes (PMN) after stimulation with phorbol myristate acetate or latex beads. PMN from patients and normal individuals were supranormally activated when resuspended in plasma from these patients. No such effect was seen with plasma from a patient with C2D but with no evidence of SLE, from patients with SLE but not C2D, from patients with C1q or C8 deficiency, from C4-deficient guinea pigs, or NZB-NZW mice. Because oxygen-derived free radicals may cause joint or tissue damage, C2D patients who have or develop this activity in their plasma may be more prone to SLE or other collagen-vascular diseases.

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

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.