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.

Determination of normal human fetal immunoglobulin M levels.

Immunoglobulin M (IgM) levels were measured in 198 cord blood samples from 192 apparently normal pregnancies from 24 weeks of gestation to term. Simple linear regression analysis yielded a standard curve for IgM development during pregnancy showing a 0.5 mg/dl increase in IgM per week of gestation. This curve allows the comparison of fetal IgM levels from pregnancies considered to be at risk for intrauterine infection.

Lymphoproliferative disorder of granular lymphocytes: nine cases including one with features of CD56 (NKH1)-positive aggressive natural killer cell lymphoma.

Twelve cases of large granular lymphocytosis were examined by flow cytometry and Southern blot analysis to correlate immunophenotypic and molecular genetic markers with clinical features. Nine cases fulfilled clinical criteria for the lymphoproliferative disorder of granular lymphocytes, and eight of these demonstrated the molecular features of T-cell-type lymphoproliferative disorder of granular lymphocytes including surface expression of CD3, expression of one or more natural killer (NK)-cell antigens (CD11b, CD16, or CD57), and clonal rearrangement of both the T-cell receptor beta- and gamma-chain-joining genes. One of these cases demonstrated coexisting clonal rearrangement of the immunoglobulin heavy-chain-joining genes, but none demonstrated kappa-light-chain-joining gene rearrangement. The eight T-cell lymphoproliferative disorder of granular lymphocytes cases all lacked expression of the NK antigen CD56 (NKH1). In contrast, the other case of lymphoproliferative disorder of granular lymphocytes rapidly evolved into an aggressive NK-cell lymphoma which did not express CD3, did express CD56, had germline T-cell receptor gene configurations, and had multiple clonal chromosomal abnormalities. This case demonstrated nasal cavity and cutaneous tumor infiltrates consistent with previously described CD3-negative, CD56-positive NK-cell lymphoma of the upper aerodigestive tract. Three cases of transient large granular lymphocytosis demonstrated germline T-cell receptor gene configurations. This study demonstrates the usefulness of Southern blot analysis and flow cytometry in characterizing proliferations of large granular lymphocytes. The transformation of a single case into an aggressive NK-cell lymphoma with blastic morphology and tissue infiltration was associated with a fatal outcome.

Detection of beta-2 transferrin in otorrhea and rhinorrhea in a routine clinical laboratory setting.

A simple, straightforward, and rapid method for the detection of beta-2 transferrin in otorrhea and rhinorrhea that can be used in a routine clinical laboratory is described. The beta-2 transferrin was detected by agarose gel electrophoresis of the fluid on Beckman Paragon equipment, followed by pressure transfer to a nitrocellulose membrane and then incubation with enzyme-labeled antitransferrin antibody and substrate. The procedure was fast (3.5 h) and sensitive (detected as little as 1 microgram/ml) and required only 3 microliters of fluid. Beta-2 transferrin was detected in cerebrospinal fluid diluted up to eightfold. No special training or expertise was needed, and all equipment and procedures used are commonly available in a routine clinical laboratory.

Quantitative criteria for clonality in the diagnosis of B-cell non-Hodgkin's lymphoma by flow cytometry.

Fifty-three cases of B-cell non-Hodgkin's lymphoma (BNHL) and 41 cases of non-BNHL lesions were retrospectively evaluated for the quantitative features of restricted surface light chain expression, pan B-cell antigens, pan T-cell antigens, and T-helper and T-suppressor phenotype using flow cytometry. Decision limit analyses were applied to multiple quantitative indices of immunophenotype to establish criteria for the detection of clonal proliferation associated with BNHL or non-BNHL conditions. Two data expressions (percent population and relative ratios) were simultaneously analyzed. The percent population measures were amenable to parametric analyses; the ratio data were amenable to nonparametric analyses only. Acceptable diagnostic specificity and sensitivity were obtained using decision limits of 75% kappa light chain and 65% lambda light chain for the detection of clonal proliferations associated with BNHL. Comparable diagnostic criteria for light chain ratios of 3:1 and 2:1 were similarly confirmed for cases of kappa clonal and lambda clonal proliferations, respectively. Neither the percent of B-cells present nor the ratio of T-helper cells to T-suppressor cells were of utility in the diagnosis of BNHL. These data confirm the numerical criteria for clonality previously obtained by cell counting studies of immunocytochemical preparations and characterize quantitative criteria for aid in the diagnosis of BNHL based on restricted surface light chain expression as measured by flow cytometry.

Effects of prednisone on human immunodeficiency virus infection.

Prednisone is used to treat a number of complications of the human immunodeficiency virus (HIV) infection, but there is little information about the immunologic consequences of such therapy. We gave prednisone to six boys with hemophilia who were infected with HIV in order to study its effects on their clinical status, serum immunoglobulins, lymphocyte populations, and serum HIV antigen concentrations. Most patients had clinical improvement while taking prednisone; platelet counts increased in five, weight increased in four, and adenopathy abated in two patients during the study. Serum IgG concentrations declined in all patients, but there were no significant changes in serum IgA or IgM concentrations. There was no consistent or significant change in the number of percentage of CD3, CD4, CD8, or CD19 lymphocytes. HIV antigen was present in the serum of one patient, and its concentration did not change during the study. None of the patients became HIV antigen seropositive during the study. These results indicate that prednisone administration was associated with clinical improvement without deleterious immunologic effects when given to children with HIV infection.

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 seroprevalence of cytomegalovirus among Virginia State prisoners.

A prevalence serosurvey of adult male prisoners entering the Virginia State Prison was conducted to evaluate the epidemiology of cytomegalovirus within this population. Four hundred and forty-five (97%) of 459 male inmates provided serum for analysis and 427 completed a detailed demographic questionnaire. Sera were tested for cytomegalovirus by passive latex agglutination and 64% were reactive. Multivariate discriminant analysis showed an independent association of seropositivity with age, non-white race, and a history of gonorrhea. There was no apparent contribution from admitted homosexual contact though this may have been under-reported. There was no correlation of seropositivity with intravenous drug use or with the length or number of prior incarcerations. Prisoners possess the same correlates for cytomegalovirus seropositivity as the general adult population; past imprisonment did not independently contribute to cytomegalovirus seropositivity.

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.

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.

Lymphocyte subpopulations of plateletapheresis products collected with the Fenwal CS-3000 Cell Separator.

To determine whether any lymphocyte subset is preferentially harvested and transfused as a consequence of plateletapheresis with the Fenwal CS-3000 Blood Cell Separator, the proportions of lymphocyte subpopulations in platelet concentrates were compared to their proportions in the donors' peripheral venous blood immediately prior to platelet collection. There was no difference in the proportion of B cells (surface immunoglobulin positive), T cells (OKT3 positive), helper/inducer T cells (OKT4 positive), suppressor/cytotoxic T cells (OKT8 positive), and natural killer cells (Leu 7 positive) in the donors' peripheral venous blood and the plateletapheresis product. Thus, although previous studies have demonstrated the ability to separate lymphocyte subpopulations by density centrifugation and velocity sedimentation, plateletapheresis with the CS-3000 harvests the lymphocyte subpopulations studied in the same proportions in which they circulate in donors' peripheral venous blood.

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.

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.

Lymphocyte subsets in normal bone marrow.

Bone marrow aspirates and biopsies from ten normal donors were stained directly with monoclonal antibodies specific for lymphocyte, monocyte, and myeloid antigens, and were analyzed by flow cytometry. To avoid cell loss, lymphocytes were not specifically isolated prior to staining. T cells comprised 46% of aspirate lymphocytes and 22% of biopsy lymphocytes. Further, the Leu-3:Leu-2 ratio of bone marrow T cells was below 1.0. B cells comprised 8% to 11% of bone marrow lymphocytes in both aspirates and biopsies, and there was a substantial percentage of cells in the lymphocyte window that was negative for all B and T cell markers. The lymphocyte window had very little myeloid contamination; however, when the myeloid window was examined, staining was greater than 90%.

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.

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.