B. J. M. Zegers

Purine Nucleoside Phosphorylase Deficiency Associated with Selective Cellular Immunodeficiency

We studied a 15-month-old girl who had normal T-cell and B-cell immunity at birth, after which a gradual decrease in T-cell immunity developed. This selective cellular immunodeficiency was inherited as an autosomal recessive trait: two older sisters had the same immunodeficiency. Adenosine deaminase activity was present in erythrocytes and lymphocytes of the patient, parents and a healthy brother. Purine nucleoside phosphorylase activity was not found in the patients erythrocytes and lymphocytes (the parents and brother had intermediate values, indicating that the enzyme deficiency too was inherited as an autosomal recessive trait). Analysis of serum and urine from the patient and of serum from her two deceased sisters showed high levels of inosine and guanosine in addition to hypouricemia and hypouricosuria. The bone marrow was megaloblastic, and the blood hypochromic microcytic. The patient had spastic tetraparesis. Intoxication of the T lymphocytes after birth by metabolic products may explain the progressive cellular immunodeficiency.

Regulation of B Cell Activity in Man: Role of T Cells

Abbreviations: AC adherent cells cig cytoplasmic immunoglobulin(s) FcyR membrane receptor for the Fc-fragment of IgG ¥cfiR. membrane receptor for the Fc-fragment of IgM He haemocyanine Helix pomatia OA ovalbumin OA-DNP12-11 ovalbumin conjugated with the hapten DNP (ratio 1 to 12-15) PBL peripheral blood lymphocytes PFC plaque-forming cell PNP purine nucleoside phosphorylase PWM pokeweed mitogen SCID severe combined immune deficiency SE sheep erythrocytes TDL tonsil-derived lymphocytes Th T heiper (cell or cells) ThFw T helper factor originating from antigen-stimulated T cells stimulated for 24 h ThFiio T helper factor originating from antigen-stimulated T cells stimulated for 120 h Ts T suppressor (cell or cells) TSB, T suppressor activator cell Tscff T suppressor effector cell TSF24 T suppressor factor originating from antigen-stimulated T cells cultured for 24 h TsFijo T suppressor factor originatingfromantigen-stimuIatedTceUscuUuredforl20h TSpr. T suppressor precursor cell Ty cell T lymphocyte expressing FcyR T^ cell T lymphocyte expressing Fc^R To T lymphocyte without demonstrable Fc/iR and FcyR

Kappa-chain deficiency. An immunoglobulin disorder.

Since kappa-chain deficiency is an unusual condition, we studied the clinical and laboratory findings in a patient with this deficiency. The patient had cystic fibrosis with concurrent malabsorption, diabetes mellitus and IgA deficiency. The serum levels of IgM and IgG were 0.85 and 7.22 mg per milliliter, respectively. Kappa type IgM and IgG was not present in serum and external secretions; gamma, mu and lambda chains were probably polyclonal in character. Antibodies against kappa chains were not detected in either the patient or the mother. Plasma cells containing kappa-type immunoglobulins were absent in jejunum samples and bone marrow; kappa-chainbearing B lymphocytes could not be detected in blood and bone marrow. The serum of one of the patients sisters contained trace amounts of kappa-type immunoglobulins. The patient displays a complete absence of kappa-type immunoglobulins, probably owing to a genetic defect.

Serum immunoglobulins in healthy children and adults levels of the five classes, expressed in international units per millilitre

Serum levels of IgM, IgG, IgA, IgD, and IgE were determined in serum samples of 270 healthy Dutch children (aged 4-13 years) and of 30 healthy Dutch adults, the amounts being expressed in International Units per millilitre. Special attention is given to the IgD and IgE results, since the IgM, IgG, and IgA levels in mg per 100 ml of these sera and their implications have already been reported. In the childrens sera the occurrence of relatively high IgD and IgE levels was frequently observed, whereas the adult group did not show excessive variation in this respect. The mean IgD levels found for adult males and females are 21 I.U./ml and 24 I.U./ml, respectively; the mean IgE levels for the same groups are 68 I.U./ml and 88 I.U./ml, respectively. The mean IgD and IgE levels in the children of each year group were usually higher than those of each of the juvenile groups and the mean level of the adult group was not statistically significant. A statistically significant influence of sex and season on the IgD and IgE levels could not be demonstrated in this material either. Three of the 270 childrens sera showed an exceptionally low IgA content. In two of these cases the serum was sampled and studied a second time after an interval of four years, when the IgA deficiency proved to be still present. The IgE levels in the sera of these healthy IgA-deficient children were normal, whereas the presence of IgD could not be demonstrated.

Role of CR2 in the human adult and neonatal in vitro antibody response to type 4 pneumococcal polysaccharide

A number of studies have indicated that the complement receptor type 2 (CR2), which is the receptor for C3d, a degradation fragment of the complement component C3, regulates B lymphocyte activation and growth. Early reports have described that C3 regulates T cell-dependent (TD) antibody responses. The involvement of CR2 in the antibody response to T cell-independent type 2(TI-2) antigens was investigated because neonatal B cells, which are unresponsive to TI-2 antigens both in vivo and in vitro, express a significantly decreased level of CR2 as compared to B cells of adult donors. We utilized type 4 pneumococcal polysaccharide (PS4) as a model TI-2 antigen. In order to study the relationship between CR2 and the response to PS4, B cells were costimulated with PS4 and monoclonal antibodies (MAb) to CR2. HB5 and OKB7 anti-CR2 monoclonal antibodies enhanced the in vitro response of adult B cells to PS4, as measured in a PS4-specific spot-forming cell assay. Neonatal B cells could only be induced to respond to PS4 using high concentrations of OKB7 anti-CR2 MAb. The 8-mercaptoguanosine (8MGuo), an agent that can overcome the in vitro unresponsiveness to PS4 of neonatal B cells, increased CR2 expression on adult and neonatal B cells. Furthermore, 8MGuo synergizes strongly with anti-CR2 antibodies in augmenting the anti-PS4 antibody response. Data presented in this report provide evidence of CR2 involvement in the antibody response to PS4 and that the neonatal B cell unresponsiveness to TI-2 antigens may be due to the decreased expression of CR2.

GENE DELETION AND GENE DUPLICATION WITHIN THE CLUSTER OF HUMAN HEAVY‐CHAIN GENES: SELECTIVE ABSENCE OF IgG SUB‐CLASSES

Individuals with selective absence of IgG1 and IgG2 were discovered by testing for allotypes and isotypes of the respective sub‐classes. These individuals were homozygous for sub‐class deleted Gm‐Am haplotypes, as shown by allotype studies in two families (Gm‐;…;g;A2ml/Gm‐;n;b;A2ml and Gm‐;n;b;A2ml/Gm‐;…;b;A2ml) and by a population study of New Guineans (Gm fa;‐;b;A2m2). The individuals with IgGl sub‐class deficiency showed elevation of IgG2, IgG4 and in particular of IgG3.

Erythrocyte Metabolism in Purine Nucleoside Phosphorylase Deficiency after Enzyme Replacement Therapy by Infusion of Erythrocytes

Purine nucleoside phosphorylase deficiency is associated with a severely defective T-cell immunity. A patient with purine nucleoside phosphorylase deficiency was treated with transfusions of irradiated erythrocytes and plasma. This resulted in a remarkable correction of the metabolic disturbances in the patient. The urinary excretion of inosine, deoxyinosine, guanosine, and deoxyguanosine decreased, whereas uric acid excretion as well as serum uric acid concentration increased. It could be shown that the enzyme activity of the circulating erythrocytes correlated inversely with the urinary excretion of nucleosides and directly with the excretion of uric acid. As a consequence of the therapy, several glycolytic intermediates of the erythrocytes were increased, especially 2,3-diphosphoglycerate. The high 2,3-diphosphoglycerate level caused a shift to the right of the oxygen dissociation curve (P50 = 32.9 mm Hg). The immunological status of the patient showed definite improvement after the enzyme replacement therapy.

An abnormal form of purine nucleoside phosphorylase in a family with a child with severe defective T-cell-and normal B-cell immunity

1. Purine nucleoside phosphorylase and adenosine deaminase (ADA) were studied in normal red blood cells and lymphocytes and in the cells of a family with a child with a defective T-cell-and normal B-cell immunity. 2. In the propositus no purine nucleoside phosphorylase (NP) activity could be detected in her red cells and lymphocytes, while the ADA activity was somewhat increased. The NP activities of the father, mother and brother of the propositus are in the heterozygote range. The decreased activity of NP was not only found for the substrate inosine but also when guanosine or xanthosine were used as substrate. The mode of inheritance is autosomal recessive. 3. With starch gel electrophoresis no NP activity could be detected in the patients haemolysate. The electrophoretic patterns of NP from the father, mother and brother of the patient seem to be the same as for normal NP with six bands of NP activity. 4. The nucleoside phosphorylases of the father, mother and brother of the patient were characterized by an increased KM for the substrate inosine, normal pH optimum and a decreased heat stability.

The serum igg subclass levels in healthy infants of 13–62 weeks of age

The levels of IgG1, IgG2, IgG3, and IgG4 were determined in serum samples of 160 infants aged 13--62 weeks, and of their mothers. In addition the serum IgM, IgG, IgA, and IgD levels of the infants are presented. The results show that IgM, IgG1, and IgG3 slightly increase during the first year of life, whereas IgG2, IgG4, IgA, and IgD hardly do. This difference in the development of the various immunoglobulin isotypes reflects differences in the terminal maturation of subsets of B-lymphocytes into plasma cells. About 50% of the infants of this age had no detectable IgG4 and three children had no IgG2. These observations indicate that longitudinal investigations are needed in children suspected of a IgG2 or IgG4 subclass deficiency. No statistically significant influence of sex on the IgG subclasses could be demonstrated in these infants.

A patient with purine nucleoside phosphorylase deficiency: Enzymological and metabolic aspects

1. Enzymological and metabolic data in a patient with nucleoside phosphorylase (NP) deficiency are described. 2. Incubation of intact NP-deficient red cells with [14C]adenosine showed a rapid uptake and conversion to inosine. Almost no radioactivity was incorporated in the adenosine nucleotides and no hypoxanthine labeling could be detected. 3. Incubation with [14C]inosine resulted in a rapid conversion to IMP in the normal intact red cells but in an accumulation of inosine in the medium with the erythrocytes of the patient, proving again that a NP deficiency is present. 4. The high PRPP level found may result from impaired consumption due to lack of substrates for the salvage enzyme HGPRT. 5. Incubation with [14C]hypoxanthine and [14C]adenine showed that normal HGPRT and APRT activities were present in the NP-deficient red cells. 6. In serum and urine of the patient the levels of inosine and guanosine were considerably increased, while the serum and urinary levels of uric acid were very low. In the two deceased sisters NP deficiency was also strongly suggested by analyses of the serum purines, of stored deep frozen samples.