Depressed cell-mediated immunity in patients with primary intracranial tumors. Characterization of a humoral immunosuppressive factor.

Tumor immunity in patients with primary intracranial tumors was assessed in relation to the general status of host immunocompetence. Lymphocyte sensitization to tumor-specific membrane antigens was demonstrated by the proliferative response of lymphocytes in the presence of autochthonous tumor cells. Paradoxically, one-half of the patients could not be sensitized to a primary antigen, dinitrochlorobenzene; existing delayed hypersensitivity was also depressed, as measured by skin tests and lymphocyte transformation in response to common antigens. A heat-stable factor in patients' sera blocked cell-mediated tumor immunity. In addition, these "enhancing" sera consistently suppressed the blastogenic response of autologous and homologous lymphocytes to phytohemagglutinin and to membrane antigens on allogeneic cells in the one-way mixed lymphocyte culture. When patients' leukocytes were washed and autologous plasma replaced with normal plasma, reactivity in the mixed lymphocyte culture increased to normal values. In vitro immunosuppressive activity in patients' plasma or sera correlated with depressed delayed hypersensitivity. After removal of the tumor, suppressor activity disappeared. IgG fractions of patient sera contained strong immunosuppressive activity. These data suggest that the suppressor factor may be an isoantibody elicited by the tumor that also binds to receptors on the lymphocyte membrane. In addition to specifically blocking cell-mediated tumor immunity, enhancing sera may broadly depress host immunocompetence.

Ventriculojugular shunt nephritis with Corynebacterium bovis. Successful therapy with antibiotics.

A patient with hydrocephalus and a ventriculojugular shunt presented with acute nephritis, nephrotic syndrome (proteinuria 10 g/24 hours), decreased complement levels, circulating immune complexes and diminished creatinine clearance (41 ml/min). Seven blood cultures grew Corynebacterium bovis. A renal biopsy specimen revealed mesangiocapillary glomerulonephritis by light microscopy, and thickened glomerular basement membranes with areas of increased granular density by electron microscopy. Immunofluorescent examination of the biopsy specimen demonstrated 2+ granular glomerular basement membrane deposits of immunoglobulin M (IgM), with trace third component of complement (C-3), fourth component of complement (C-4) and immunoglobulin G (IgG). Rabbits immunized with C. bovis produced a line of partial identity in agar with patient serum against a sonicate of C. bovis. Indirect fluorescein staining of the biopsy specimen with the rabbit antiserum demonstrated 1+ granular glomerular basement membrane deposits. Potassium thiocyanate microelution of sections prior to examination markedly diminished staining with antihuman antiserum, but did not affect staining with rabbit antiserum. Following initial therapy with intravenous penicillin for six weeks the bacteremia cleared, serum complement levels returned to normal, proteinuria decreased and creatinine clearance increased. A relapse occured four weeks later with decreased complement levels, increased proteinuria and decreased creatinine clearance. Blood cultures were again positive for C. bovis. Following therapy with erythromycin and rifampin, the bacteremia cleared and there was a sustained improvement of all parameters. To our knowledge, this is the first time an association has been noted between C. bovis ventriculojugular shunt infection and glomerulonephritis. These findings support the potential role of C. bovis as an etiologic agent in human renal disease and further define the immune complex nature of shunt nephritis.

Comparison of five methods for the analysis of the light chain type of monoclonal serum IgM proteins.

Five methods for the analysis of the light chain type of monoclonal IgM proteins present in serum at low concentration were compared in terms of sensitivity, feasibility for use in a clinical laboratory, technical effort required, and cost. These studies indicated that methods based on the removal of IgG from serum were the most appropriate for routine use.

Phenotype analysis of lymphocyte subsets in normal human bone marrow.

T-lymphocyte subsets in marrow aspirates taken from normal donors were analyzed with the use of monoclonal antibodies and flow cytometry. Peripheral blood specimens from the same donors were analyzed concurrently and served as controls. Eighty-six percent of CD8-positive cells from marrow expressed the pan-T-cell markers CD2 and CD3, compared with 91% and 81%, respectively, of CD8-positive cells from peripheral blood. Virtually all CD8-positive marrow lymphocytes, however, were negative for CD11b as opposed to peripheral blood, where 18% exhibited positivity for this marker. Further, a higher percentage of marrow CD8-positive T-lymphocytes expressed HLA-DR (15.4%) than did those from peripheral blood (4%). Marrow CD4-positive subsets did not differ substantially from those of peripheral blood. These results support the concept of a selective homing of CD8-positive T-cells to the marrow and indicate that CD8-positive T-cells in the marrow are effector T-cells.

The hydrolysis of human IgG with subtilisin.

Small amounts of subtilisin (0.1 mg. per ml.) in the absence of cysteine will instantaneously hydrolyze "enzyme-sensitive" IgG globulins. This procedure permits the identification of antibodies in whole serum and myeloma globulins in the ultracentrifuge with simplicity and speed unmatched by other technics.

Hepatitis B virus serology. Loss of antibody to surface antigen.

A retrospective review of hepatitis B serologies from 6,686 patients was conducted to determine the incidence of loss of antibody to hepatitis B surface antigen (anti-HBs) with retention of antibody to the core antigen of hepatitis B virus (anti-HBc) activity. In a subgroup of 48 multiply tested patients who were presumed to have resolved their acute or subacute hepatitis B virus infection, 9 (19%) were found to have lost anti-HBs while retaining anti-HBc activity.

Hepatocytic globules in end-stage hepatic disease: relationship to alpha1-antitrypsin phenotype.

Hepatic explant specimens from 171 patients with cirrhosis were examined to determine the incidence of periodic acid-Schiff (PAS)-positive diastase-resistant globules (PDRGs) in end-stage hepatic disease and whether the globules bear a specific relationship to the alpha1-antitrypsin (A1AT) phenotype or to causes of hepatic disease other than A1AT deficiency. PAS-positive diastase-resistant globules were detected in 17 (10%) of the hepatic explant specimens, and the globules in all of these cases were strongly immunoreactive for A1AT. In the 17 patients with PDRGs, the cirrhosis was attributed preoperatively to A1AT deficiency (3 patients), ethanol abuse, viral hepatitis, or both (10 patients), cryptogenic cirrhosis (3 patients), and autoimmune hepatitis (1 patient). The A1AT isoelectric phenotypes classified according to the protease inhibitor (Pi) nomenclature for 16 of these patients were as follows: Pi ZZ (3 patients), Pi SS (1 patient), Pi MZ (8 patients), and Pi MM (4 patients). Because PDRGs were seen in a variety of A1AT phenotypes, serum electrophoretic analysis, not histologic examination, is required for the correct diagnosis of an A1AT abnormality. Furthermore, although PDRGs were seen in a variety of hepatic diseases, the majority of patients with globules had an undetected A1AT abnormality. Accordingly, on identification of hepatocytic PDRGs, the clinician should be alerted to the possibility of an unsuspected A1AT abnormality even in the presence of other causes of hepatic disease.

Cohort study of human immunodeficiency virus (HIV) antibody testing among patients receiving long-term dialysis at a university hospital.

In a longitudinal study to determine the seroprevalence of antibody to the human immunodeficiency virus (HIV) and the natural history of a positive enzyme immunoassay (EIA) result we followed a cohort of 98 patients receiving long-term dialysis. Eight patients (8.2%) in the cohort had a positive EIA and a negative Western blot test result. The EIA-positive results of all patients seroconverted to negative during follow-up. No illness suggestive of HIV infection developed in any of the patients. Significantly associated with a false positive EIA were prior renal transplantation, transfusions during the months just before the positive EIA result, and a greater number of lifetime transfusions before the positive test result. We confirm that routine HIV screening of patients receiving long-term dialysis is associated with a high rate of false positive EIA results and conclude that such testing is unnecessary in the absence of established risk factors for HIV infection.

Gamma heavy chain disease: report of a case associated with trisomy of chromosome 7.

A case of gamma heavy chain disease is reported in a 52-year-old white male who presented with fever and generalized lymphadenopathy. A lymph node biopsy showed malignant lymphoma. A partial transient response was obtained with cyclophosphamide, vincristine, prednisone, and doxorubicin. He died 3 months after diagnosis from disease progression and infectious complications. Chromosome analysis of cells from an involved lymph node showed the presence of trisomy 7. Chromosome abnormalities have been reported in three of ten previously published cases of gamma heavy chain disease. Trisomy of chromosome #7 has not previously been reported.

Seroepidemiology of hepatitis D (delta agent) and hepatitis B among Virginia State prisoners.

A serosurvey of hepatitis D (HDV) and hepatitis B (HBV) was conducted in an asymptomatic population of newly incarcerated prison inmates in Virginia. Of 459 men entering the prison, 445 provided both sera and demographic and personal information. Six (1.3%) had antibody to HDV (anti-HDV). Evidence of past infection with HBV was found in 146 (32.8%); 9 (2.0%) were positive for HBV surface antigen (HBsAg). HBV seropositivity correlated with intravenous drug abuse, nonwhite race, and tattoos acquired in prison. Sera obtained after an interval of seven to ten months revealed seroconversion to anti-HDV in one of two HBsAg-positive men who admitted to parenteral drug use while incarcerated. Because he had been incarcerated elsewhere for more than one year before entering this prison, it is concluded that HDV transmission occurred in prison. The association of HDV infection with progression to chronic active and fulminant hepatitis suggests that serologic surveillance of HBsAg-positive inmates may be indicated for identification of possible HDV index cases.

Effects of dexamethasone therapy on serum immunoglobulin levels and peripheral cell counts in women undergoing conservative benign gynecologic surgery.

Twenty women undergoing conservative gynecologic surgery were treated perioperatively with high-dose dexamethasone (DEX) for 72 hours. Serum immunoglobulin levels (IgG, IgM, IgA) and peripheral cell counts, including lymphocyte subpopulations, were measured in each patient during the immediate preoperative and postoperative periods as well as 5 to 8 weeks later. These results were compared with the levels of a control population of 16 women undergoing conservative, non-pregnancy-related gynecologic surgery for benign conditions. There was a decrease in the level of all serum immunoglobulins measured in the immediate postoperative period in both groups, with a greater statistical decrease in the control group (P less than 0.001). statistically significant transient alterations in the peripheral cell counts occurred in the DEX group, as compared with the control group (P less than 0.001). The use of DEX was not associated with any postoperative infectious morbidity.

Quantitative effects of intraperitoneal 32% dextran 70 on serum immunoglobulin levels in women undergoing gynecologic surgery.

The use of intraperitoneal 32% dextran 70 results in quantitative postoperative changes in human IgG and IgA levels that are statistically different from the alterations induced by the surgical procedure alone.

In vitro production of monoclonal human rheumatoid factors.

Lymphocytes from rheumatoid arthritis patients have been fused with drug-marked human myeloma cell lines and hybrids selected which secrete rheumatoid factors. Limiting dilution and soft agarose plates have been used to clone these hybrids, but so far stable clones have not been obtained. Epstein-Barr virus has been used to transform lymphocytes, and clones which secrete rheumatoid factors have been obtained.

Detection of oligoclonal IgG bands in cerebrospinal fluid by isoelectric focusing on thin layer agarose gels.

The detection of oligoclonal bands of IgG in cerebrospinal fluid by isoelectric focusing is a useful aid in the diagnosis of multiple sclerosis. At present, this test is performed on polyacrylamide gels which are technically very difficult to prepare and are not suitable for the routine laboratory. A technique is described for isoelectric focusing on thin layer agarose gels which permits oligoclonal banding detection in the routine laboratory in four hours. The advantages of this procedure over isoelectric focusing on polyacrylamide gels include: (a) it takes less time (four hours compared with at least 24 hours); (b) it requires less specimen (2.5 ml compared with reference laboratory requirements of at least 4 ml); (c) it is technically easier to perform; and (d) it can be done routinely in the clinical laboratory.

The detection of oligoclonal IgG bands in unconcentrated cerebrospinal fluid by isoelectric focusing on thin-layer agarose gels and silver staining.

Isoelectric focusing on thin-layer agarose gels with silver staining was used to detect oligoclonal bands of IgG in unconcentrated cerebrospinal fluid (CSF). Comparison of this procedure with Coomassie blue staining for the detection of oligoclonal bands in CSF yielded concordant results; CSF samples were either positive or negative for oligoclonal bands by both techniques. The silver-staining procedure was capable of detecting oligoclonal bands in CSF samples with IgG levels as low as 2 mg/dL, which was 200 ng/10-microliter sample. False-positive results were not detected in CSF samples with either a high initial IgG level or a low initial IgG level that required concentration before analysis, even by silver staining. This procedure has several advantages over conventional assays: it is fast, needing only five hours; it uses only 10 to 15 microliter of sample; it is easy to perform; and it can be done routinely in the clinical laboratory.

Quantitation of serum immunoglobulins.

This article reviews critically the current "state of the art" in the quantitation of immunoglobulins in serum and other body fluids. The methodologies reviewed include (a) those occurring in solution, for example, automated immunoprecipitin and laser nephelometric techniques, rate analysis techniques and radioimmunoassays; (b) techniques involving radial diffusion in agarose gels, with and without secondary development steps; and (c) those techniques involving solid supports, such as fluoroimmunometric assays and solid phase radioimmunoassays. The theory of each technique is presented with an analysis of the strengths and weaknesses, particularly sources of error, and the techniques are compared in terms of accuracy, precision, sensitivity, test cost, equipment cost, feasibility for use in different laboratory settings, and ease of handling. Problems associated with the antisera used, with standards, and with quality controls are discussed and solutions suggested.

Use of kappa and lambda chain quantitation for the detection of immunoglobulin abnormalities in serum.

Quantitation of kappa and lambda light chains was investigated as an alternative to immunoelectrophoresis or immunofixation in the analysis of sera for abnormal immunoglobulins. Normal values for the ratios of total immunoglobulins to total light chains, and for total kappa chains to total lambda chains, were established in 104 control sera and were 1.070 +/- 0.074 and 1.770 +/- 0.337, respectively. Protein electrophoresis plus quantitation of immunoglobulin heavy and light chains provided sufficient data for the analysis of 79% of apparent normal sera, 76% of apparent abnormal sera which did not have monoclonal components, 73% of abnormal sera which contained monoclonal components, and 53% of pediatric sera. However, this protocol failed to detect monoclonal components known to be present in two sera. These data indicate that this analytic approach cannot replace the subjective analysis of sera by immunoelectrophoresis or immunofixation.