Karyotype of Friend virus-induced mouse erythroleukemia cells

Banded karyotypes were prepared for three independent sublines of clone 745A, a Friend virus-induced erythroleukemia cell line originally produced by Dr. C. Friend. Each subline had the same chromosome complement, with a near-diploid number of chromosomes, two-thirds of which appeared normal. The remaining one-third were structurally rearranged marker chromosomes, including eight whose origins could be determined from their banding patterns. A fourth subclone of line 745A, under different selection pressure, showed changes in five marker chromosomes but no changes in the normal chromosomes, suggesting that the markers contained unstable sites, perhaps viral integration sites. Another Friend virus-induced erythroleukemia cell line, FSD-1/F4, developed independently by Dr. W. Ostertag, had a near-diploid number of chromosomes, about one-fourth of which were abnormal chromosomes. The only similarity between these markers and those in subclones of 745A, other than participation of some of the same chromosomes in centric fusion, was the inclusion of the proximal two-thirds of chromosome 15 in one of the markers of each line. This adds to the number of mouse tumor cells or cell lines in which No. 15 is altered or duplicated. In FSD-1/F4 cells there was a striking increase in the size of the secondary constriction of one of the two chromosomes 19, suggesting that the rRNA genes on this chromosome had been amplified.     

Cryptosporidiosis of the human small intestine: a light and electron microscopic study

Intestinal infection by the coccidian parasite Cryptosporidium is a well-recognized condition in immunocompromised hosts and in some normal persons. The authors studied a patient with acquired immunodeficiency syndrome and cryptosporidiosis of the small intestine. The parasite inhabits the microvillous brush border of the intestinal epithelium and must be carefully sought on light microscopic examination of intestinal biopsy specimens. Characteristic life cycle stages are observed on electron microscopy. The absence of significant light microscopic alterations of the villous architecture in this patient's biopsy specimen and in other cases suggests that other factors, such as toxin elaboration by cryptosporidia or other organisms, may be involved in the pathogenesis of diarrhea. Abnormal aggregation of lysosomes at the apices of intestinal epithelial cells may reflect ineffective host phagocytic mechanisms.     

Monoclonal antibody identification of mononuclear cells in endomyocardial biopsy specimens from a patient with rheumatic carditis

A 17-year-old woman with rheumatic carditis underwent endomyocardial biopsy both prior to and following treatment with prednisone and aspirin. Frozen sections from the endomyocardial biopsy specimens were studied with monoclonal antibodies by an indirect immunofluorescence technique to define the composition of the inflammatory infiltrate in the myocardium and to determine whether the composition of the infiltrate is distinctive and diagnostically useful. The specimen from the initial biopsy contained a heterogeneous infiltrate composed of T lymphocytes, macrophages, B lymphocytes, and mast cells. T lymphocytes predominated, and the ratio of T-helper to T-cytotoxic/suppressor cells was 2.0. Following treatment the overall cellularity of the infiltrate was diminished, but the infiltrate remained heterogeneous; T cells predominated, and the T-helper to T-cytotoxic/suppressor ratio was reversed, to 0.59. The composition of the inflammatory infiltrate in this case of rheumatic carditis distinguishes it immunologically from other "idiopathic," presumably virus-associated, forms of myocarditis.     

Hepatitis B virus and myocarditis

Myocarditis may be a serious extrahepatic complication of hepatitis. In this fatal case of serologically documented hepatitis B viral hepatitis, acute myocarditis was present, with histologic features consistent with a viral pathogenesis. Hepatitis B surface antigen was demonstrated by immunoperoxidase methods in small intramyocardial vessels, suggesting that hepatitis B virus infected the heart. The resulting inflammatory heart disease may have been caused either directly, by virus infecting the myocardium, or indirectly, by an immune-mediated mechanism.     

Kaposi's sarcoma following chemotherapy for testicular cancer in a homosexual man: demonstration of cytomegalovirus RNA in sarcoma cells

A 35-year-old white Jewish homosexual man who had undergone surgery and chemotherapy for an embryonal carcinoma of the testis subsequently developed Kaposi's sarcoma. The neoplasm involved the skin as well as visceral tissues. Tissue derived from a biopsy specimen of one of the skin lesions was used in the in situ hybridization technique for the detection of genetic material. Cytomegalovirus messenger RNA was identified in the neoplastic Kaposi cells in the skin. The significance of this finding is discussed.     

Enhancement of idiotype-anti-idiotype T cell interactions by monoclonal OKT11A antibody

We have demonstrated that OKT11A, a mouse monoclonal antibody recognizing the E receptor on human T lymphocytes, strongly enhances the autologous mixed lymphocyte culture (AMLC) response induced by T lymphoblasts, while inhibiting the AMLC response induced by B lymphoblasts as well as the T cell response to alloantigens and soluble antigens. This antibody seems to enhance the idiotype-anti-idiotype interaction between resting and activated T lymphocytes.     

The relationship of family history of alcoholism to primary affective disorder.

One hundred sixty-eight patients with primary affective disorder were studied regarding history of alcohol-related problems. Alcohol-related problems were identified in 19 patients. We found that the morbid risk for alcoholism was significantly increased among relatives of male patients with drinking problems as compared to relatives of male patients without drinking problems. Our data suggest that psychiatric illness in relatives of probands with severe bipolar illness tends to be affective disorder tends to include alcoholism plus affective disorder.     

X-linkage in bipolar affective illness: Perspectives on genetic heterogeneity,pedigree analysis and the X-chromosome map

The search for genetic markers is a powerful strategy in psychiatric genetics. The present article examines four areas relevant to discrepancies among X-linkage studies in bipolar affective disorder. These are questions of ascertainment, analytic methods, the X-chromosome map and genetic heterogeneity. The following conclusions are reached: (a) Positive linkage findings cannot be attributed to ascertainment bias or association between affective illness and colorblindness. (b) The possibility that falsely positive linkage results were obtained by using inappropriate analytic methods is ruled out. (c) Reported linkages of bipolar illness to colorblind and G6PD loci are compatible with known map distances between X-chromosome loci. Linkage to the Xg antigen remains uncertain. (d) The discrepancy among the various data sets on affective illness and colorblindness is best explained by significant linkage heterogeneity among pedigrees informative for the two traits.     

Angiosarcomas arising in edematous extremities: immunostaining for factor VIII-related antigen and ultrastructural features

Immunostaining for Factor VIII-related antigen was seen in deparaffinized sections from 19 of 20 postmastectomy angiosarcomas and from four of four sarcomas that arose in chronically edematous tissue unrelated to breast carcinoma. Staining was also seen in sections from two malignant hemangioendotheliomas, four capillary hemangiomas, and one granulation tissue specimen. Sections from two lymphangiomas were immunonegative for Factor VIII-related antigen in the endothelium of lymphatic channels, whereas staining was observed in the surrounding normal blood vessels. Electron microscopic study of four postmastectomy angiosarcomas disclosed ultrastructural features (fenestrae, intense pinocytotic activity, cell junctions, and Weibel-Palade bodies) supporting the blood vascular endothelial nature of the neoplastic cells. It is concluded that a neoplastic blood vessel component is present in sarcomas that arise in chronically edematous tissues. It is questionable whether a lymphatic component is also present. These tumors, therefore, should be regarded as angiosarcomas rather than lymphangiosarcomas.     

The significance of cystic hygroma in fetuses

Cystic hygroma of the neck was observed in seven spontaneously aborted, severely autolyzed female fetuses. Four of five karyotyped cases had the 45,XO karyotype; one was a normal female, 46,XX. Diagnostic features associated with monosomy X in fetuses, which were observed in six of these cases, include large cystic hygromas, generalized edema, edematous chorionic villi with scalloped borders and trophoblastic inclusions, and, possibly, a small heart. In contrast, the single euploid fetus in this series had a small cystic hygroma, no hydrops, and chorionic villi of normal size. These features may differentiate cystic hygromas associated with the XO karyotype, which are not associated with recurrence risk, from those associated with a familial syndrome.     

Distinct but overlapping T helper epitopes in the 37-58 region of SSX-2

Because of their specific expression in tumors of different histological types, the products of the SSX genes are important candidate targets for development of cancer vaccines. We have previously identified two immunodominant SSX-2-derived T cell epitopes recognized by HLA-A2-restricted CD8+ T cells (SSX-2 41-49) and HLA-DR11-restricted CD4+ T cells (SSX-2 45-59), respectively. In this study, we report the identification of an HLA-DR3-restricted epitope mapping to the 37-51 region of SSX-2, overlapping both previously identified epitopes. As about one fifth of individuals from several major ethnic groups express HLA-DR3, the identification of this epitope significantly increases the percent of patients that are expected to mount specific CD4+ T cell responses following vaccination with peptides in this region of SSX-2. Retrieval of multiple overlapping epitopes in a defined region of SSX-2 protein suggests the presence of a "hot spot" for T cell recognition that may prove sufficient for the induction of immune responses.     

Immunological cross-reactions of the major internal protein component from "slow" viruses of sheep.

The major p27 polypeptides of the sheep “slow” viruses, visna, maedi, and progressive pneumonia, were compared by radioimmunoassay. The immunologic relatedness among these viral proteins was determined from the competition of homologous- and heterologous-disrupted virions with iodinated visna virus p27 for visna antisera. The p27 antigens of visna virus and maedi virus were indistinguishable from each other and were partially related to p27 of progressive pneumonia virus.     

Standardized Semi-structured Psychosocial Evaluation before Hematopoietic Stem Cell Transplantation Predicts Patient Adherence to Post-Transplant Regimen

In patients undergoing stem cell transplantation (SCT), nonadherence has potential for significant medical impact and potentially life-threatening complications. No study thus far has demonstrated an effective way to predict adherence in SCT recipients. A structured rating scale, the Stanford Integrated Psychosocial Assessment for Transplantation (SIPAT), has been shown to predict psychosocial outcomes and medical morbidity in solid organ transplant recipients. We assessed the SIPAT in SCT recipients. We hypothesized that the SIPAT rating would be associated with nonadherence to the post-SCT regimen. We retrospectively studied SCT recipients who had psychiatric evaluations with the SIPAT before SCT. The primary outcome was nonadherence, defined a priori as at least 1 life-threatening nonadherence event in the first 6 months post-transplant. Association of the SIPAT with outcomes was evaluated by logistic regression, and an optimal cutoff score was determined using a receiver operating characteristic curve. Of 85 patients (mean age 47 years; range, 18 to 74 years), 56 (66%) were male, and 43 (50.5%) received autologous SCT. Eighteen (21%) patients were nonadherent. The SIPAT rating, treated as a continuous variable and controlling for autologous versus allogeneic SCT, was significantly associated with nonadherence (per 1 point; odds ratio [OR], 1.162; P< .0001). Allogeneic SCT also conferred a significantly increased risk of nonadherence (OR, 14.184; P= .005). Multivariate analysis stratifying for allogeneic versus autologous transplantation and controlling for age, sex, and disease confirmed an independent association between the SIPAT score and nonadherence. A cutoff score of 18 provided optimal specificity (89.6%) and sensitivity (55.6%) for nonadherence. Nonadherence rates were 58.8% and 11.8% for subjects with SIPAT ratings of 18 and above or 17 and below, respectively (relative risk = 4.98, P < .0001). Psychosocial risk as quantified by the SIPAT correlated with SCT recipients' adherence to the post-transplant regimen, suggesting that this instrument can contribute to medical risk stratification models. Further study should evaluate long-term mortality data and the effects of intervention on psychosocial risks.     

Glycophorin A on normal and leukemia cells detected by monoclonal antibodies, including a new monoclonal antibody reactive with glycophorins A and B

A new hemagglutinating monoclonal antibody, MoAb31, detected glycophorins A and B in Western blots. Results with enzyme-modified erythrocytes indicated the MoAb31 determinants were sialic acid dependent, and resided on glycophorin A on the trypsin-resistant, ficin-sensitive segment, and on glycophorin B on the ficin-sensitive segment. Another new monoclonal antibody, MoAb36, detected the Wrb antigen, located on the non-glycosylated segment of glycophorin A near its insertion into the lipid bilayer. Immunofluorescent staining of normal hematopoietic and leukemia cells with these and other monoclonal antibodies to glycophorin A demonstrated glycophorin A on erythroid cells only. Cytofluorograph analysis showed the majority of cells of the erythroleukemia cell lines K562 and HEL expressed glycophorin A, as indicated by reactivity with the monoclonal glycophorin A antibodies R10, R18, 6A7 and 10F7. However, reactivity with monoclonal antibodies to glycosylated determinants (MoAb31 and R1.3) and to the non-glycosylated segment near the membrane insertion (MoAb36, and R7.1) was reduced or absent. Expression of "missing" glycophorin A antigens on K562 and HEL could not be induced using a variety of chemical and biologically active modifiers. We conclude that glycophorin A of erythroleukemia cell lines K562 and HEL differs from glycophorin A at the surface of normal, mature erythrocytes with respect to reactivity with monoclonal glycophorin A antibodies.     

Development of a protein a enzyme immunoassay for use in screening hybridomas

A protein A enzyme immunoassay has been developed which is effective in the rapid screening of hybridomas. The detecting reagent is a stable alkaline phosphatase-protein A conjugate, prepared by a simple, inexpensive, single-step procedure. The assay requires small amounts of antigen to coat microtiter wells (less than 25 μg/well) and can be evaluated visually or with a spectrophotometer. It compares favorably in sensitivity to a protein A-radioimmunoassay and enjoys considerably lower backgrounds than that popular screening procedure.     

Isolation and characterization of influenza virus recombinants with high and low neuraminidase activity. Use of 2-(3'-methoxyphenyl)-n-acetylneuraminic acid to identify cloned populations

Two antigenically identical HON2 influenza virus recombinants derived from A/England/42/72 (H3N2) and A/PR8/34 (HON1) were separated from a mixed population by the use of a staining procedure specific for neuraminidase. This procedure employing an artificial neuraminidase substrate (2-(3′-methoxyphenyl)-N-acetylneuraminic acid, MPN) permitted the identification of deeply stained (MPN⁺) and poorly stained (MPN^?) clones of virus in clone 1-5C-4 cells. Upon isolation and purification of the 2 variants, they were found to have eightfold differences in the ratios of neuraminidase/hemagglutination activity. Analysis by polyacrylamide gel electrophoresis demonstrated that the differences in neuraminidase activity reflected differences in the amount of neuraminidase per virion. The MPN^? variant which had low neuraminidase content replicated to equal titer in eggs and formed larger plaques than MPN⁺ variant on clone 1-5C-4 cells. Attempts to isolate clones of MPN^? virus from wild type A/England/42/72 virus have not been successful so far.