The majority of mouse spontaneous rosette-forming cells of thymic origin belong to the Ly-2+ subset (original) (raw)
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Journal of Experimental Medicine, 1972
By using the two criteria (a) high density of immunoglobulin determinants on the cell surface and (b) presence of receptors for C'3 on the cell surface for defining bone marrow-derived lymphocytes, it is indirectly shown that all or at least a major population of human thymus-derived lymphocytes under certain conditions will form nonimmune rosettes with sheep red blood cells (SRBC). Almost all thymocytes tested from two different donors formed rosettes. The SRBC rosettes are not formed by virtue of immunoglobulin receptors and form only around living cells. Positive bivalent ions are required for rosette formation since EDTA will block rosette formation. Sodium iodoacetate will also block rosette formation demonstrating the dependence on an intact glycolytic pathway. Rosette formation is temperature dependent and will not appear at 37°C. Trypsin treatment of lymphocytes will abolish their SRBC-binding ability which cannot be restored by treating them with fresh donor serum or fe...
Dependence of mouse thymocyte-erythrocyte rosette formation on complete identity at class-l-MHC
Journal of Cellular Physiology, 1991
Mouse thymocytes and erythrocytes form rosettes when incubated together at 4°C. The frequency is much higher when the thymocytes and erythrocytes are MHC-identical. If the indolizidine alkaloid swainsonine (SW) is present during rosette formation at concentrations of 1 p g h l (5.7 pM) or greater, rosette formation between MHC-identical pairs is inhibited to levels comparable to those observed for MHC-different pairs; rosette formation by MHC-different pairs is not affected. This was confirmed by examining 17 different MHC-identical combinations (9 completely syngeneic and 8 differing in non-MHC genes) and 13 MHC-different combinations (3 of these identical everywhere except at MHC). A SW-inhibitable component of rosette formation was observed only when thymocyte and erythrocyte were completely identical at MHC. Thus F,-parent pairs behaved as if allogeneic, although both F,-F, and parent-parent had a SWinhibitable rosetting component. Similarly, inbred strains only partially MHCidentical (BIO.BR-BlO.A, BlO.D2-BlO.A) behaved as if allogeneic. The SWinhibitable component of rosetting could be partially but significantly blocked by including monoclonal antibodies against Thy-I, and antiLCD4 plus anti-CD8 (together but not separately); anti-class-I-MHC produced some inhibition of marginal significance. Monoclonal antibodies against class-I-MHC, LFA-1, and CD3 did not block. Pretreatment of erythrocytes with neuraminidase greatly reduced the SW-inhibitable component of rosetting. The SW effect would appear to be due to a direct interaction of SW with a cell surface structure involved in syngeneic rosette formation rather than the known ability of SW to block the processingof N-linked sugar structures. The results are consistent with cell surface lectins and cell surface sugars playing a role in rosette formation.
Rosette formation between murine lymphocytes and erythrocytes. A new locus in the H-2 region
The Journal of …, 1979
Histocompatibility is necessary for many collaborative and aggressive cell interactions. The sharing of alleles at certain loci of the H-2 region on chromosome 17 of the mouse is essential for optimal interaction of lymphocytes with each other (1-7), with macrophages (8, 9), and with target cells in certain types of cytotoxic killing by T lymphocytes (10-17). This evidence indicates that lymphocytes can and must recognize products of the H-2-chromosomal region for optimal responses to take place. Several curious observations which bear on the general question of self-recognition by lymphocytes have appeared in the recent literature. Koskimies and M~ikel~i (18) reported that T-cell-deficient mice made stronger anti-hapten responses to conjugates of syngeneic erythrocytes than to conjugates of allogeneic or xenogeneic erythrocytes. These findings, together with others demonstrating that an appreciable fraction of murine lymphocytes form rosettes with autologous, but not with isologous, erythrocytes (19-20), support the existence on at least some immunocytes of a receptor for a self-marker which is expressed on erythrocytes. Because this form of self-recognition is likely to be of significance for a complete understanding of the mechanisms of lymphocyte activation, it was of interest to further investigate the phenomenon of autologous rosette formation. The present study confirms the phenomenon, establishes that the receptor in question is expressed on both T and B lymphocytes, and that it is not immunoglobulin in nature. Moreover, the use of recombinant strains of mice indicates that this form of self-recognition is associated with a marker coded by a new locus in the H-2-region mapping between H-2G and H-2D. Materials and Methods Mice. Various strains of mice were purchased from Simonsen Laboratories, Gilroy, Calif. Some recombinant strains were kindly provided by Dr. J. Stobo
Rosette formation with goat erythrocytes. A marker for human T lymphocytes
Clinical and experimental immunology, 1978
We demonstrate the use of goat erythrocytes in a rosette procedure for the classification of human lymphocytes. The population is almost perfectly overlapping with the lymphocytes which form rosettes with sheep red blood cells. 70'2 ± 7-5% of peripheral lymphocytes form rosettes with goat erythrocytes and less than 1% of these cells have surface immunoglobulins. Enrichment of goat rosette-forming cells results in a population with an increased percentage of both goat and sheep rosettes. This population retains activity to the T-cell mitogens Con A and PHA, while the cells depleted of goat rosettes have greatly diminished responses to these same mitogens. Tonsil and spleen lymphocytes form 50-2 -4 68% and 24% ofgoat rosettes respectively, while peripheral blood lymphocytes from patients with CLL rarely form goat rosettes. Cell lines maintained in vitro rosetted with goat cells in a parallel fashion to sheep cells. Thus T-cell lines, such as Molt-3, which form rosettes with SRBC also rosette with GRBC, while sheep rosette-negative lines, i.e. Molt-4, are negative for both erythrocytes. B-lymphoid cell lines were negative, as were several lymphoma cell lines. There was a slight variation in the binding of goat cells, depending on the source of the goat. Thus, as in sheep rosettes, some animals were better sources than others, although all the animals tested formed rosettes.
Formation of mouse erythrocyte rosettes by human lymphocytes. A B-cell marker
26 5 + 866% of human peripheral lymphocytes form rosettes with mouse erythrocytes. There is a significant correlation between the numbers of mouse erythrocyte rosetteforming (MERF) lymphocytes and immunoglobulin-bearing cells. By density gradient centrifugation the MERF cells can be separated from the other lymphocytes. Studies on the isolated MERF cells indicate that every MERF lymphocyte is an Igbearing cell, but they do not possess sheep erythrocyte-binding receptors and cannot be stimulated with phytohaemagglutinin. Accordingly, the MERF lymphocytes are regarded as B cells.
Journal of Immunological Methods, 1976
Purified human B and T lymphocytes were obtained by rosetting HPL with AET-SRBC or MRBC and separating the non-rosetted from the rosetted cells on Ficoll-Hypaque gradient. 92 ± 3% of the purified B cells were fluorescent positive for MBIg and 95 ± 2% of the purified T cells rosetted with AET-SRBC. 69 ± 5% of the B cells and 61 ± 8% of the T cells present in the unfractionated HPL were recovered in the purified fractions. * Abbreviations used in this paper: AET, 2-aminoethylisothiouronium bromide hydrobromide; AET-SRBC, SRBC treated with AET; AHG, aggregated human gamma globulin; B cell, bursal equivalent; EAC, complement coated sensitized sheep erythrocytes; EAC1-5 rab, immune adherence positive EA coated with C1-C5 from rabbit serum deficient in C6; EAC3d m°, immune adherence negative EA coated with C1-C3 from mouse serum deficient in C5 ; HPL, human peripheral lymphocytes; MBIg, membrane bound immunoglobulin; MEM, minimum essential medium; MRBC, monkey red blood cells; SRBC, sheep red blood cells; T cell, thymus derived; VBS, veronal-buffered saline.
“Active” T cells in guinea pig peripheral blood lymphocytes
Clinical Immunology and Immunopathology, 1981
The ability to form rosettes with rabbit red blood cells (RRBC) has been shown to be characteristic of T but not B cells from guinea pigs. In an attempt to devise an assay system to measure the guinea pig equivalent of human "active" T cells, we have investigated the effects of various conditions on RRBC binding by guinea pig lymphocytes, including incubation time, RRBC:lymphocyte ratio, presence or absence of guinea pig serum or fetal calf serum in the incubation medium, and pretreatment of RRBC with neuraminidase or 2-aminoethylisothiouronium bromide (AET). Without incubation and at a 100: 1 ratio of RRBC to guinea pig lymphocytes, about 35% of guinea pig peripheral blood lymphocytes bound RRBC, similar to the number of "active" T cells in human peripheral blood as measured by rosette formation with sheep red blood cells. Cold incubation increased the number of rosetting cells to over SO'%, presumably the total number of T cells. A relative decrease in the number of RRBC diminished rosetting only at ratios of 1O:l or less. Pretreatment of RRBC with neuraminidase slightly increased, whereas AET pretreatment drastically reduced the number of rosettes. Increasing concentrations of guinea pig serum in the medium up to 100% slightly inhibited rosetting. These results suggest the presence of two T-cell populations in guinea pig peripheral blood, equivalent to the human "active" and "total" rosette-forming T-cell subpopulations.