Anne B. Wilson

Rosette-Formation between Guinea Pig Lymphoid Cells and Rabbit Erythrocytes – a Possible T-Cell Marker

A proportion of guinea pig lymphoid cells have an affinity for rabbit erythrocytes which is demonstrable by a rosette-forming technique. In normal adult guinea pigs, this reaction has been shown to oc

Comparison of the direct antiglobulin rosetting reaction with the mixed antiglobulin rosetting reaction for the detection of immunoglobulin on lymphocytes

Abstract A procedure for the Direct Antiglobulin Rosetting Reaction (DARR) is described in which Ig anti-immunoglobulin is linked by chromic chloride to trypsin-treated ox red cells. Such anti-immunoglobulin-linked red cells do not form E or EA rosettes with human lymphocytes yet are very sensitive in forming rosettes with SmIg positive lymphocytes. The DARR on fresh live lymphocytes gives the same percentage reactivity as the Mixed Antiglobulin Rosetting Reaction (MARR) on formaldehyde-fixed lymphocytes. The two reactions have been compared in four different species. Formaldehyde treatment markedly reduces the reactivity of lymphocytes in the DARR.

Interleukin-2-induced production of interferon-γ by resting human T cells and large granular lymphocytes: Requirement for accessory cell factors, including interleukin-1

Between 5 and 20% of normal human lymphocytes were found to synthesize interferon-gamma (IFN-gamma) in primary cultures with recombinant interleukin-2 (rIL-2). After 22 hr, IFN-gamma-producing cells included CD5+ T lymphocytes, CD16+ large granular lymphocytes (LGL), and a population of CD5-, CD16- blast cells. Only a small proportion (0-7%) of IFN-gamma-synthesizing cells expressed HLA-DR. The production of IFN-gamma by all rIL-2-responding lymphocyte subsets was shown to require the presence of DR+ accessory cells, probably including nonadherent, esterase-negative monocytes and/or dendritic cells. Accessory cell function in lymphocyte preparations depleted of DR+ cells, or in purified (greater than or equal to 95%) suspensions of LGL, was fully replaced either by addition of 2% autologous, adherent monocytes or by monocyte culture supernatant. The activity of monocyte supernatant was greatly reduced by treatment with antiserum specific for human interleukin-1 beta (IL-1 beta), although a combination of rIL-1 beta and rIL-2 failed to stimulate IFN-gamma production in DR- lymphocytes. These results indicate that rIL-2-induced IFN-gamma synthesis in both T cells and LGL requires the synergistic activity of IL-1, and possibly of one or more other monokines, as yet unidentified.

Antibody-dependent cellular cytotoxicity in the guinea pig: The role of the Kurloff cell

Abstract In the guinea pig, the killer cell in in vitro ADCC assays, is found to be the Fc receptor-bearing Kurloff cell. This killer Kurloff cell is under hormonal control, estrogen treatment significantly increasing the Kurloff cell numbers in blood, spleen, and thymus, and markedly augmenting the lytic capacity of these lymphoid compartments. The cytotoxic killer lymphocyte, if present, appears to be a minor effector cell in guinea pig ADCC. Selective lymphocyte subpopulation and Kurloff-cell depletion procedures reveal that the killer Kurloff cell may also possess a variety of other membrane markers (T + , C3 + , Ig + ). The particular membrane profile of a cytotoxic Kurloff cell is determined by its lymphoid site of residence and the hormonal status of the animal.

Receptors for Antibody-Opsonic Adherence on the Eosinophils of Guinea Pigs

Eosinophils have recently been implicated in antibody-dependent cell-mediated damage to schistosomula. Because of this, eosinophils of the guinea pig have been examined for surface receptors capable of giving antibody opsonic adherence; a rosetting reaction has been used. The eosinophils were shown to possess Fc receptors for homologous immunoglobulin. No selective difference between IgG1 and IgG2 was observed. In marked contrast to macrophages, guinea pig eosinophils failed to show opsonic adherence to red cells sensitized to a comparable degree with rabbit antibody. With red cell antibodies made in the pig, however, the reciprocal situation held, namely opsonic adherence was stronger with eosinophils than with macrophages.

Increased affinity of guinea pig thymocytes and thymus-dependent lymphocytes for papain-treated rabbit erythrocytes compared to untreated erythrocytes

The affinity of guinea pig thymocytes and peripheral T lymphocytes for rabbit erythrocytes was found to be enhanced following treatment of the erythrocytes with papain. By increasing the numbers of rosette-forming cells and giving stronger, more stable rosettes, this procedure increases the usefulness of the reaction as a T cell marker in guinea pigs.

Antigen S1, encoded by the MIC1 gene, is characterized as an epitope of human CD59, enabling measurement of mutagen-induced intragenic deletions in the AL cell system

S1 cell membrane antigen is encoded by the MIC1 gene on human chromosome 11. This antigen has been widely used as a marker for studies in gene mapping or in analysis of mutagen-induced gene deletions/mutations, which utilized the human-hamster hybrid cell-line, AL-J1, carrying human chromosome 11. Evidence is presented here which identifies S1 as an epitope of CD59, a cell membrane complement inhibiting protein. E7.1 monoclonal antibody, specific for the S1 determinant, was found to react strongly with membrane CD59 in Western blotting, and to bind to purified, urinary form of CD59 in ELISAs. Cell membrane expression of S1 on various cell lines always correlated with that of CD59 when examined by immunofluorescent staining. In addition, E7.1 antibody inhibited the complement regulatory function of CD59. Identification of S1 protein as CD59 has increased the scope of the AL cell system by enabling analysis of intragenic mutations, and multiplex PCR analysis of mutated cells is described, showing variable loss of CD59 exons.

Complement components on human lymphocytes

Abstract Lymphocytes isolated from normal human EDTA-blood were tested for the presence of cell membrane-bound complement components by reaction with specific antibodies coupled to trypsin-treated ox erythrocytes. The highest proportions of positively reacting cells were found to have C4 and β1H-globulin, which were present on 11 and 9% (mean values) of lymphocytes, respectively. Other complement components, C1q, C3, and C6, were detected on approximately 3% lymphocytes. C4 was also demonstrated on tonsil and lymph node lymphocytes. All of the cells carrying complement were C3-receptor-bearing, Ig-positive, B lymphocytes, a proportion of which also had receptors for IgG-Fc.

Artefactual variations in the B and T subpopulations of rabbit blood lymphocytes depending on method of isolation, and blocking of C3 receptors due to in vitro activation of complement.

Marked differences were found in the proportions of lymphocyte subpopulations in rabbit peripheral blood depending on the techniques used for the purification of lymphocytes. Rosette-forming reactions were used to find the numbers of T lymphocytes, Ig-bearing cells and cells with receptors for C3 or IgG-Fc. Some of the methods used for lymphocyte separation altered the relative numbers of T and B lymphocytes, through a disproportionate loss of T cells. Other changes were due to in vitro activation of complement detectable by the presence of C3 on the lymphocyte cell-membrane and causing partial blocking of C3 receptors. Highest yields of lymphocytes were obtained from defibrinated blood treated with carbonyl iron to remove phagocytes and methyl cellulose to sediment erythrocytes. This procedure was accompanied by in vitro activation of complement, with the consequences mentioned. Complement activation was inhibited by taking the blood into either EDTA or citrate. As EDTA was cytotoxic for rabbit T lymphocytes, citrate was considered best although the resulting lymphocyte suspensions were contaminated with up to 25% granulocytes and monocytes owing to the inhibition of carbonyl iron uptake by the prior exposure to citrate.

Reaginic antibodies on basophils in grass pollen allergy, as shown by rosette-formation.

Antibodies to grass pollen allergens have been demonstrated on basophils of individuals with hay fever by means of a rosette-forming reaction with allergen-coated erythrocytes. The antibodies on the b