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Spontaneous rosette formation and rosette inhibition assays in patients with squamous cell carcinoma of the head and neck.

Peripheral blood lymphocytes from patients with squamous cell carcinoma (SCC) of the head and neck were studied by spontaneous lymphocyte rosette and rosette inhibition (RI) assays prior to treatment. The patients were clinically staged and the results of the assays compared with the clinical stage of the disease. The percentage of T-lymphocytes as determined by the spontaneous lymphocyte rosette test was significantly lower (p less than .01) for the patient group when compared with a normal population. Patients with stage I and II disease did not differ significantly from controls. Individuals with stage III or IV disease, however, had significantly lower T-lymphocyte counts. The tumors were histologically graded as well, moderately well, or poorly differentiated SCC. Patients with poorly differentiated neoplasms had significantly lower T-cell counts. The RI assay (using horse anti-human thymocyte globulin to inhibit rosette formation) was abnormal in many of the patients but did not appear to be a more sensitive in vitro measure of cell mediated immunity in these patients. Performing both tests detected more patients with cellular immunologic incompetence than either one alone.

Adolescent↗

Rosette formation between human lymphocytes and sheep erythrocytes. Inhibition of rosette formation by specific glycopeptides.

Rosette formation with unsensitized sheep erythrocytes is a characteristic of human thymus dependent lymphocytes. Release of glycopeptides from the sheep erythrocyte by trypsin reduces rosette formation. These tryptic glycopeptides inhibit rosette formation by untrypsinized sheep erythrocytes; this suggests that rosetting is mediated by erythrocyte surface glycopeptides. To investigate the molecular nature of this interaction, we examined the abilities of various model compounds to act as haptenic inhibitors of rosette formation. Inhibition is given by glycopeptides bearing oligosaccharide units rich in sialic acid, galactose, N-acetylglucosamine, and mannose linked to asparagine residues through glycosylamine bonds. Among compounds tested, fetuin glycopeptide is most effective, but human transferrin glycopeptide and human erythrocyte glycopeptide I also inhibit rosette formation. Other compounds including human erythrocyte glycopeptide II, human IgG glycopeptide, lacto-N-neotetraose, 3'- and 6'-sialyllactose show no significant inhibition. Neither sialic acid, galactose, manose, nor N-acetyl-glucosamine alone inhibits rosette formation. Stepwise degradation of fetuin glycopeptide established the galactose residues as important determinants of inhibitory activity. Fetuin glycopeptide blocks rosette formation when added to a suspension of human lymphocytes and sheep erythrocytes or when preincubated with human lymphocytes, but not when preincubated with sheep erythrocytes. Studies of the binding of [3H] fetuin glycopeptide to normal lymphocytes demonstrate 7.5 x 10(6) saturable binding sites per cell. No saturable binding of this compound to sheep erythrocyte membranes is observed. Compared to normals, lymphocytes from patients with chronic lymphatic leukemia demonstrate decreased fetuin glycopeptide binding with a mean of 0.9 x 10(6) sites per cell. This decreased binding correlates with the impaired ability of these cells to form rosettes. The data suggest that fetuin glycopeptide inhibits rosette formation by binding to the thymus-dependent cell where competition occurs with sheep erythrocytes for specific lymphocyte surface receptors.

Animals↗

Abnormal spontaneous rosette formation and rosette inhibition in lung carcinoma.

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.

Aged↗

T lymphocyte colony-forming capacity of patients with immunodeficiency diseases: relationship of colony formation to E rosette formation and lymphocyte proliferation.

The formation of T lymphocyte colonies was studied in 30 normal subjects and 12 patients with suspected abnormalities in immune function. The mean number of colonies per plate in normal subjects was 1.159 +/- 411 which represented a plating efficiency of 0.5-1.0%. When individual cells from colonies of these normal subjects were studied for membrane markers, greater than 90% were E rosette-positive and less than 1% were positive for surface immunoglobulins. Blood lymphocytes obtained from all 12 patients showed diminished colony-forming capacity when compared to normal subjects with a range of 0-311 colonies. Six patients had less than 50 colonies/plate. Colony formation was diminished in some patients who had normal E rosette formation and lymphocyte proliferation in liquid culture. Because of these discrepancies it appears that colony formation is not a direct reflection of E rosette formation and lymphocyte proliferation. Evaluation of T lymphocyte colony-forming capacity may prove useful as an additional in vitro assessment of lymphocyte function.

Cell Division↗

Cells involved in autologous rosette formation in mice. I. Two different subpopulations of thymocytes regulating autologous rosette formation.

In the studies on mouse thymocyte subpopulations--non-glass-adherent (NAL) cortisone-sensitive (HCS), and glass-adherent (GAL) cortisone-resistant (HCR) thymocytes the existence of precursors for helper and suppressor cells of the autologous rosette formation was demonstrated. A proline-rich polypeptide (PRP), an immunoregulator induced differentiation of helper and suppressor cells from NAL and GAL cells, respectively. These cells elevate or inhibit formation of autologous rosette by thymocyte subpopulations in vitro. Helper activity was abolished by treatment with monoclonal anti-Lyt 1,1 antibodies and suppressor activity was totally inhibited after treatment with monoclonal anti-Lyt 2,1 antibodies.

Animals↗

Rosette formation in osteosarcoma.

Rosette formation is a rare, recently reported variation in osteogenic sarcoma and is thought to be associated with a poor prognosis. We report two cases of rosette forming osteosarcoma, one with poor response and other with total necrosis following chemotherapy. Pathologists should be aware of rosette formation in osteosarcoma to avoid misdiagnosis as other rosette forming tumors of bone especially PNET/Ewings sarcoma. In our opinion rosettes in an osteosarcoma should be documented both from a differential diagnostic point of view and also to elucidate definitive prognostic implications.

Adult↗

Cell electrophoretic characterization of peripheral blood lymphocyte subpopulations enriched by rosette formation, from normal individuals and CLL patients.

Peripheral blood lymphocytes from healthy subjects and patients with chronic lymphatic leukemia (CLL) were isolated and their subpopulations enriched through formation of spontaneous rosettes with sheep or mouse red blood cells, respectively. Electrophoretic measurements were performed in unseparated as well as in fractionated cell populations. Normal blood lymphocytes showed two clearly distinguishable populations of different electrophoretic mobilities. After separation by SRBC rosette formation the rosette-forming cells could be identified as high mobility cells. CLL lymphocytes showed in most cases an unimodally distributed cytopherogram, the mean electrophoretic mobility being intermediate between the low and high mobility cells of control persons. After separation through mouse erythrocytes rosette formation these cells contained two cell fractions differing in their electrophoretic mobility: a fraction of slower mouse rosette-forming cells and a fraction of the non-MRFC which contained mainly cells of higher mobility that could be identified as enriched T cells. These both fractions showed unimodal distributions. This study shows that CLL lymphocyte subpopulations can be further characterized by surface charge density.

Animals↗

Inhibition of T gamma rosette formation by the sera from patients with systemic lupus erythematosus. Relation to T gamma-specific antilymphocyte antibody.

The cytotoxic activities of lupus sera were measured against IgG Fc-receptor-bearing T lymphocytes (T gamma cells) and T lymphocytes lacking this receptor [T gamma (-) cells], and the activities were compared with the inhibitory activities of the sera for the formation of rosettes (T gamma rosettes) between T lymphocytes and ox erythrocytes sensitized with IgG antibody. The cytotoxic activities of the sera against both T gamma and T gamma (-) cells well correlated with their T gamma rosette inhibitory rates. Also, the cytotoxic activities after the removal of IgM antibodies strongly correlated with the inhibitory rates. Among them, the highest correlation was observed between IgG T gamma-specific cytotoxic antibodies and T gamma rosette inhibitory rates of the sera. Gel filtration, ultracentrifugation, pepsin digestion, and reduction and alkylation of the sera revealed that main inhibitory activities were contained in IgG fractions. These results suggested that IgG T gamma-specific antibody suppressed T gamma rosette formation and might contribute to the reduction of T gamma cell number in patients with systemic lupus erythematosus.

Antibody Specificity↗

Rosette formation in Plasmodium falciparum isolates and anti-rosette activity of sera from Gambians with cerebral or uncomplicated malaria.

The ability of Plasmodium falciparum-infected red blood cells (RBC) to form spontaneous erythrocyte rosettes was studied in 130 fresh isolates from Gambian children with cerebral or uncomplicated malaria from August to November 1990. All isolates (24 of 24) from patients with cerebral malaria formed rosettes, but only 61 of 106 isolates from children with uncomplicated malaria formed rosettes. The mean rate of rosette formation in isolates from children with cerebral malaria (28.3%) was significantly greater than that in isolates from children with uncomplicated malaria (8.5%). Giant rosettes were more frequently formed in isolates from patients with cerebral malaria than in those from patients with uncomplicated malaria. Sera of children with cerebral disease generally lacked anti-rosette activity, while many sera from children with uncomplicated malaria showed strong anti-rosette activity when tested against the patients' ow parasites. Some sera that were devoid of autologous rosette-disrupting activity were able to disrupt rosettes formed in other isolates, indicating the presence of different rosette formation mechanisms. Forty percent (6 of 15) of the sera from patients with cerebral malaria caused microagglutination of the patients' own uninfected and infected RBC, while only 10% (3 of 31) of sera from children with uncomplicated disease caused microagglutination. The ability of infected RBC to bind to melanoma cells grown in vitro did not differ between patients with cerebral or uncomplicated malaria. The results of this study, taken in conjunction with our previous findings, establish a strong association between rosette formation in P. falciparum-infected RBC and cerebral malaria.

Adolescent↗

Mixed glioneuronal tumour of the fourth ventricle with prominent rosette formation.

We describe three unusual tumours characterized by a mixture of glial and neuronal differentiation, involvement of the posterior fossa and formation of rosettes. Mixed glial-neuronal tumours of the posterior fossa are rare and poorly described neoplasms. However, several distinctive entities have appeared in the literature over recent years under a variety of different names. Our cases demonstrate the morphological features of the 'rosette-forming glioneuronal tumour of the fourth ventricle', a recently identified tumour characterised by its unique location, neurocytic pseudo-rosette formation and the presence of a low grade astrocytoma component. The long term prognosis of these tumours remains unclear. However, the clinical data available including the cases presented here, along with the histological features, suggest that these are low grade tumours with a good prognosis after surgical resection.

Adult↗

Detection of nerve growth factor binding sites on neuroblastoma cells by rosette formation.

A cloned suspension culture of mouse C1300 neuroblastoma cells bound, at 2degrees, sheep erythrocytes passively coated with nerve growth factor, with the formation of rosettes. When grown in tissue culture dishes to which they could attach, neuroblastoma cells rapidly transformed, within 48 hr emitting cytoplasmic processes some of which were several mm long. Most of the attached neuroblastoma cells formed rosettes. In contrast, normal mouse kidney cells or various murine tumor cell lines used as cell controls exhibited a poor capacity for binding nerve growth factor. Rosette formation was a specific reaction that could be prevented by pretreating cells with proteolytic enzymes, free nerve growth factor, or specific antibodies against neuroblastoma cell extracts.

Animals↗

Attachment and rosette formation by hyphomicrobia.

The processes of reversible and irreversible sorption of Hyphomicrobium sp. strain ZV580 to solid surfaces was investigated with the use of swarmer cell populations. Various compounds and physical conditions were examined for their effect on the cell-cell interactions which lead to the formation of rosette-like cell aggregates. The monosaccharides galactose and mannose were able to completely inhibit rosette formation. Concanavalin A and chloramphenicol were also able to prevent rosette formation, and both trypsin and pronase caused the disassociation of rosettes. The results indicate that the adhesin is a glycoprotein or a peptido-polysaccharide and that the flagellum may act as a vehicle for the transport of this adhesin from the cell to a surface.

Journal Article↗

Binding of mouse red blood cells (MRBC) by human thymocytes: augmentation of rosette formation by phospholipase C treatment.

The formation of rosettes with MRBC is not an exclusive property of chronic lymphocytic leukemia cells and of human B-lymphocytes. Human thymus cells and the HD-MAR T-cell line were also found to be capable of forming rosettes with MRBC. The binding of MRBC by thymocytes differed from that of sheep (SRBC), rabbit (RRBC) and human (HRBC) erythrocytes by a number of parameters: monoclonal antibodies against the E-receptor inhibited the attachment of SRBC, RRBC, and HRBC, but not of MRBC to thymus cells. The presence of EDTA had the opposite effect, abrogating only the attachment of MRBC to thymus cells but not of RBC from the other sources. Exposure of thymus cells to pronase eliminated their capacity to bind SRBC, RRBC, and HRBC, but only slightly inhibited the attachment of MRBC. Treatment with Vibrio cholerae neuraminidase enhanced the formation of rosettes with SRBC, RRBC, and HRBC, but had no effect on the formation of rosettes with MRBC. Exposure of thymus cells and of the HD-MAR cells to phospholipase C enhanced the proportion of rosettes formed with MRBC, but had no effect on the binding of other RBC. Treatment with either phospholipase A2 or phospholipase D had no such effect. The binding of MRBC by Raji cells was not increased by phospholipase C treatment. The present study indicates that the binding of MRBC by human thymus cells is mediated by receptors distinct from those involved in the binding of SRBC, RRBC, and HRBC and also from those mediating the binding of MRBC to human B-cells.

Animals↗

High-affinity erythrocyte rosette formation and inhibition in premalignant disease of the uterine cervix.

Total lymphocyte counts, total rosette formation, high-affinity rosette formation, and the rosette inhibition titer were studied in 104 patients with dysplasia, carcinoma in situ, or Stage I squamous cell carcinoma of the uterine cervix. Although no changes were detected in total lymphocytes or total rosette formation, a significant decrease (p less than 0.001) was observed in high-affinity rosette formation in patients with moderate and severe dysplasia and carcinoma in situ. The rosette inhibition titer was significantly increased (p less than 0.001) in all stages of dysplasia and carcinoma in situ; incubation of patients' serum with normal lymphocytes did not reproduce this change but significantly decreased the rosette inhibition titer of the patients' lymphocytes (p less than 0.001). No change in high-affinity rosette formation or the rosette inhibition titer was observed in patients with Stage I squamous cell carcinoma as compared to controls. These changes are postulated to represent alterations in the normal balance of T-cell differentiation during the early stages of tumor development.

Adult↗