HLA and survival in lung cancer.

HLA A and B antigens were determined in a study of 125 patients with lung cancer. Differences between antigen frequencies in cancer and control populations were determined by chi 2 analysis or Fisher's exact test. Survival data were analyzed using the Cox model for censored data. Cancer patients had an increased frequency of the antigen Aw33 (relative risk = 10.5, P less than 0.016). The Cox model (D. R. Cox, J. R. Stat. Soc. B, 34, 187, 1972) indicated that four covariates had a significant effect on mean survival time independently: the presence of A3 (P less than 0.005) and of Aw33 (P less than 0.05) increased mean survival time of the cancer population; patients with anaplastic carcinoma and stage three of any histological type of cancer had a decreased mean survival time. The determination of HLA phenotypes, cancer type, and the stage of the disease can provide the expected mean survival time of any particular patient. This could be of importance for evaluating prognosis. The effect of Aw33 and A3 on survival time may be related to HLA closely linked genes, possibly coding for resistance to the disease.

Modification of a direct enzyme-linked immunosorbent assay for the detection of immunoglobulin G and M antibodies to pneumococcal capsular polysaccharide.

In contrast to the usual indirect enzyme-linked immunosorbent assay (ELISA) method for detection of antibody responses, a modified direct ELISA technique was used to measure immunoglobulin G (IgG) and IgM responses to pneumococcal capsular types 1, 3, 9N, and 23F in humans. Individual capsular polysaccharides were covalently bound to poly-L-lysine before adsorption to the solid phase. The coupling reaction was enhanced by maintenance of a constant pH of 8.2 after the addition of all reactants. The evaluation of four diluents (phosphate-buffered saline [PBS]-Tween; PBS-Tween plus 10% fetal calf serum; PBS-Tween plus 10% bovine serum albumin; and PBS-Tween plus 20% normal goat serum) showed that the sensitivity and specificity of the assay was increased with normal goat serum (10-fold). Serum samples from 10 subjects immunized with polyvalent pneumococcal vaccine were tested by direct ELISA and by radioimmunoassay. At 4 weeks postimmunization, the ELISA method showed that IgG was the predominant antibody and that IgM responses were lower or had diminished. Isotype shifts during this period would have been undetected by the radioimmunoassay method. The changes in antibody response measured by ELISA were comparable to the radioimmunoassay results. The direct ELISA method for the detection of antipneumococcal capsular antibody has been found to be a sensitive and reproducible assay for the detection of IgG and IgM antibodies.

Major histocompatibility locus in the Arabian horse.

Combined immunodeficiency disease (CID) is a genetic disorder of T and B lymphocyte production which results in a nonfunctional immune system. It is inherited as an autosomal recessive trait and has been reported in humans and in horses of the Arabian breed. Arabian horses known to have the CID gene and horses of unknown carrier status were tested using a microlymphocytotoxicity technique. Computer chi 2 analysis distinguished six serologically defined specificities. The study of unrelated horses and a limited number of families showed that the specificities behave as codominant alleles segregating at one locus. No differences in antigen frequency was detected between the CID carriers and the random horse population.

Purification of an adenosine deaminase complexing protein from human plasma.

A protein which specifically complexes with adenosine deaminase (complexing protein) has been purified to homogeneity from human plasma. This protein was compared with complexing protein isolated from human kidney. The two proteins produce electrophoretically different forms of high molecular weight adenosine deaminase when combined with the Mr = 36,000 enzyme monomer from erythrocytes. This difference may, at least in part, be due to the greater sialic acid content of complexing protein from plasma. By other criteria, including amino acid composition, total carbohydrate content, and subunit structure, the two proteins are quite similar. In addition, plasma complexing protein shows complete cross-reactivity with anti-kidney complexing protein serum. These results suggest that plasma and kidney complexing proteins are products of the same gene.

Studies on the pathogenesis of experimental autoimmune renal tubulointerstitial disease in guinea pigs. VI. Induction of renal lesions by active or passive immunization of strain 2 guinea pigs.

It has been reported that strain 2 guinea pigs do not develop experimental autoimmune renal tubulointerstitial disease (RTD) by active or passive immunization. Under our experimental conditions strain 2 guinea pigs are susceptible to the induction of RTD. Typical moderate to severe renal lesions were seen 22 days after immunization with rabbit tubular basement membrane in complete Freund's adjuvant. Most Albany strain guinea pigs had severe RTD. Susceptibility to induction of RTD by passive transfer of antitubular basement membrane autoantibodies was studied in Albany (A) and strain 2 (2) guinea pigs, as well as in A leads to A, A leads to 2, and 2 leads to A radiation chimeras. Only some strain 2 guinea pigs had mild to severe renal lesions by day 9, but by day 19 all had typical histologic renal abnormalities. Albany guinea pigs already had moderate to severe RTD by days 9-10. Strain 2 differed from Albany recipients by a noticeable delay in the onset of lesions. Bone marrow transplants between Albany and strain 2 did not affect these susceptibility differences. A leads to 2 recipients responded like strain 2, and 2 leads to A like Albany. This indicates that the delay of onset of RTD in strain 2 is not a defect in the bone marrow-derived inflammatory cells.

Localization of an adenosine deaminase-binding protein in human kidney.

A protein which specifically complexes with adenosine deaminase (complexing protein) has been localized in human kidney. Thin sections of renal tissue were treated with rabbit anti-complexing protein serum followed by fluorescein-labeled goat anti-rabbit gamma globulin serum. Complexing protein was detected in the proximal tubules of four normal kidneys. The glomeruli in sections from one of the four normal kidneys were also positive. Lung, liver, spleen, cardiac muscle, skeletal muscle, and kidney medulla were negative by this technique. The glomeruli as well as the proximal renal tubules of two patients with combined immunodeficiency disease-adenosine deaminase deficiency and three of seven patients with kidney disease contained easily detectable levels of complexing protein.

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

A number of infants with an autosomal recessive form of combined immunodeficiency disease also lack adenosine deaminase (adenosine aminohydrolase; EC 3.5.4.4) activity in their erythrocytes. Other tissues from these infants contain only a few percent of the adenosine-deaminating activity present in corresponding normal tissue. The residual adenosine-deaminating activity in extracts from the spleen of a combined immunodeficient, adenosine deaminase-deficient patient was compared with adenosine deaminase from normal spleen. Affinity and immunoadsorbant column chromatography revealed distinct differences between the adenosine-deaminating activity in the patient's spleen and adenosine deaminase from normal spleen. The point of maximum activity and general configuration of the pH optimum curves were also different. erythro-9-(2-Hydroxyl-3-nonyl)adenine, a potent inhibitor of adenosine deaminase from normal spleen, had relatively little effect on the activity from the patient's spleen. In contrast, adenine was a better inhibitor of the activity in the patient's spleen than it was of the enzyme from normal tissue. An adenosine-deaminating activity with the same characteristics and specific activity as that in the patient's spleen was also isolated from normal spleen. These results suggest that the adenosine-deaminating activity in the spleen of this patient is not due to a mutant form of adenosine deaminase.

Studies on the pathogenesis of experimental autoimmune renal tubulointerstitial disease in guinea-pigs. IV. failure to inhibit mononuclear cell accumulation with niridazole.

Niridazole is a potent inhibitor of certain expressions of delayed-type hypersensitivity; however, its effect and specificity on other immunologically induced mononuclear cell infiltrates has not been determined. We therefore tested this drug in a model of auto-antibody-induced renal tubulointerstitial disease (RTD), which is characterized by the accumulation of lymphocytes and macrophages in the cortical interstitium. Niridazole-treated (100 mg/kg) and untreated guineapigs were injected with potent anti-tubular basement membrane serum from donors with RTD. Drug treatment was repeated 2-3 times during the course of 10 days. The characteristic renal lesions were found in both groups. However, a single dose of niridazole inhibited skin reactivity in tuberculin sensitive guinea-pigs for at least 8 days. These findings further support the contention that immunologic tissue injury in RTD does not involve mechanisms of delayed-type hypersensitivity.

Purification of human erythrocyte adenosine deaminase by affinity column chromatography.

Adenosine deaminase (adenosine aminohydrolase EC 3.5.4.4) has been purified 468,000-fold from pooled human erythrocytes. The procedure developed was used to isolate the enzyme from up to 23 liters of packed erythrocytes at one time. An easily prepared affinity column bed material employing adenosine as the ligand was used as the final step in the purification. During elution from the affinity column there was approximately a 3:1 partition of adenosine deaminase between gel bed and column buffer. There was no apparent difference in the partitioning of unresolved or partially resolved preparations of the electrophoretically different forms of the enzyme on the affinity column. Gel filtration and electrophoresis on polyacrylamide gels of increasing concentration revealed no differences in the Mr of these electrophoretically different forms. The four bands resolved by electrophoresis of the different forms on polyacrylamide gels under nondenaturing conditions yielded a single band when electrophoresis was carried out in the presence of sodium dodecyl sulfate and 2-mercaptoethanol. Partially resolved preparations of the different electrophoretic forms of adenosine deaminase also gave rise to a single band of the same mobility when electrophoresed on polyacrylamide gels under these conditions. The band had the mobility of a protein of Mr of 36,000. This Mr is approximately the same as estimated for the nondenatured enzyme.

Evaluation of adenosine deaminase and other purine salvage pathway enzymes in horses with combined immunodeficiency.

Foals with combined immunodeficiency had normal levels of purine salvage pathway enzymes, including adenosine deaminase, nucleoside phosphorylase, and xanthine oxidase.

Combined immunodeficiency disease associated with adenosine deaminase deficiency. Report on a workshop held in Albany, New York, October 1, 1973.

Fifty-five children with CID and known ADA status were studies at a workshop held in Albany, New York. Erythrocyte ADA determinations were performed in 22 of the 55 patients, 13 of whom were ADA negative. The ADA defect appears to be transmitted as an autosomal recessive trait. Some patients with CID and ADA deficiency have characteristic radiologic abnormalities of the skeleton, which are not found in other illnesses. The thymus glands of all patients with CID and ADA deficiency who could be examined have evidence of thymic involution manifested by presence of Hassall's corpuscles and differentiated germinal epithelium; this is in contrast to "classic" thymus findings in CID with normal ADA. Adenosine deaminase probably plays an important, although as yet undefined, role in lymphocyte development and/or function. The deficiency of ADA in CID is the first enzyme defect observed in a deficiency disease of specific immunity.

Combined immunodeficiency disease: an inborn error of purine metabolism.

The recently discovered association of adenosine deaminase (ADA) deficiency and combined immune deficiency (CID) has emphasized the critical role of purine salvage in the function of lymphoid tissue. Known enzymatic properties of ADA are presented. In addition, known phenotypic data and possible genetic mechanisms for the occurrence of ADA deficiency in CID are discussed. A hypothesis based on considerations of known metabolic pathways in human erythrocytes is proposed to account for the selectivity of ADA deficiency for lymphoid tissue. Finally, some inhibitors of ADA are discussed as well as some immunosuppressive agents.

Adenosine deaminase deficiency in combined immunologic deficiency disease.

Deficiency of red cell and lymphocyte adenosine deaminase (ADA) was found in children suffering from congenital combined immunologic deficiency. The parents had ADA levels intermediate between patients and controls. Complete lack of ADA activity was not found in normal subjects or in patients with a variety of other immunologic deficiency diseases.