Epstein-Barr virus induced differentiation of early B-lineage cells.
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B-Lymphocytes↗
Biomedical subjects
M D Cooper
Publications and source records attributed to M D Cooper.
The role of receptors for complement in the induction of polyclonal B-cell proliferation and differentiation.
A panel of monoclonal antibodies and ligands that bind to the CR1 or CR2 complement receptors of B cells has been used to investigate the role of these membrane molecules in regulating B-cell proliferation and differentiation. When CR2 was modulated from the surface of B cells by treatment with the HB-5 antibody and a secondary goat anti-mouse immunoglobulin antibody, Epstein-Barr virus-induced polyclonal B-cell proliferation and immunoglobulin production were inhibited by 83 and 90%, respectively. In contrast, modulation of other cell surface molecules, HB-2, B1, and the C3b receptor (CR1), or pretreatment of B cells with C3d,g (a CR2 ligand) or HB-5 antibody, alone minimally inhibited these responses. Neither the HB-5 antibody C3d,g, nor a monoclonal antibody (YZ-1) reactive with CR1 induced resting B cells to proliferate, nor did they alter anti-mu antibody-induced proliferation. Similarly, treatment with C3d,g or with the HB-5 or YZ-1 antibodies did not induce B cells to secrete immunoglobulin or affect pokeweed mitogen-induced plasma-cell formation. Whereas CR2 appears to be the functionally relevant receptor for Epstein-Barr virus on B cells, the effects of ligand interactions with CR1 and CR2 on normal B-cell proliferation or differentiation remain unidentified.
Adult↗
[18F]fluoride from a small cyclotron for the routine synthesis of [18F]2-fluoro-2-deoxy-D-glucose.
[18F]Fluoride was produced via the 18O (p, n) 18F reaction using a low volume stainless steel 18O-water target in a small cyclotron. The average 18F production rate for 60 runs was about 720 microCi/microA-min. A typical bombardment time of 20 min produced 120 mCi of 18F-fluoride. [18F]2-Fluoro-2-deoxy-D-glucose ([18F]2FDG) was prepared using the procedure developed by Tewson [J. Nucl. Med. 24, 718 (1983)] involving a protected cyclic sulfate glucose derivative as precursor. The routine synthesis and quality testing of [18F]2FDG required 1 h and, typically, yields of 30-35 mCi of greater than 95% HPLC-pure product were obtained.
Blood Glucose↗
Pathogenesis of IgA1 protease-producing and -nonproducing Haemophilus influenzae in human nasopharyngeal organ cultures.
We evaluated mucosal attachment, colonization, and invasion by Haemophilus influenzae in an experimental model of human nasopharyngeal tissue in organ culture. Nonpiliated, encapsulated, and nonencapsulated, IgA1 protease-deficient mutants of H. influenzae were compared with their isogenic IgA1 protease-producing parents. Damage to peripheral ciliary activity was first noted 6 hr after infection and was associated with sloughing of ciliated cells to which H. influenzae were not attached. Infection of organ cultures with each strain resulted in similar degrees and rates of ciliary damage. H. influenzae attached selectively to nonciliated epithelial cells or was associated with surface mucus. Later, disruption of epithelial tight junctions was observed, and clusters of H. influenzae were found between epithelial cells. Organisms were also seen within phagocytic vacuoles of mononuclear cells located above and below the basement membrane. In summary, encapsulated and nonencapsulated H. influenzae damaged the ciliary function of human nasopharyngeal organ cultures, attached to the mucosal surface, and invaded the epithelium. H. influenzae IgA1 protease, however, was not essential for the pathogenic steps observed in this human nasopharyngeal organ culture model.
Haemophilus Infections↗
Localization of gonococcal lipopolysaccharide and its relationship to toxic damage in human fallopian tube mucosa.
An experimental model using human fallopian tubes in organ culture was used to study the localization of purified gonococcal lipopolysaccharide (LPS). LPS was visualized by light microscopy with immunoperoxidase staining. Immediately after addition to fallopian tube organ cultures, gonococcal LPS aggregated on the tips of cilia. By 1 to 2 h after exposure, LPS could be seen distributed throughout the cytoplasm of ciliated and nonciliated cells in structures resembling vesicles. By 12 h, there were sloughed, ciliated cells present in the fallopian tube lumen, which had positive LPS stain on their surfaces as well as in their cytoplasm. By 24 h, LPS was distributed throughout the cytoplasm. Control experiments with rabbit oviduct organ cultures showed that LPS failed to attach, enter, or damage mucosal cells. These studies illustrate the initial localization of LPS on human mucosal cells and its uptake into the cells, which are coincident with toxicity for ciliated epithelial cells.
Animals↗
Effect of oxytocin, prostaglandin F2 alpha and reproductive tract manipulations on uterine contractility in Holstein cows on days 0 and 7 of the estrous cycle.
The influence of various treatments on cattle at the time of natural estrus vs estrus induced by prostaglandin F2 alpha (PGF2 alpha) or at d 7 of the estrous cycle were studied; the latter is when embryo transfer often is performed. Eight lactating and 25 nonlactating, normal cycling cows were tested many times while in estrus and 7 d after estrus. A balloon was positioned in the body of the uterus to record changes in intrauterine pressure following clitoral massage, cervical massage, vaginal distention, electrical stimulation of the cervix and vagina, tailhead rubbing, udder massage and the injection of oxytocin or PGF2 alpha. Blood oxytocin and intramammary pressure were measured. There were no differences between cows in estrus spontaneously or those induced, so these groups were combined. Intravenous oxytocin injections of .5, 1, 2, 4 and 15 IU increased blood levels of oxytocin. Intramammary pressure was increased by all oxytocin doses, but greater than or equal to 2 IU were required to cause substantial changes in uterine contractions. As expected, the peak contractions during control periods for cows in estrus were high, averaging 31 mm Hg vs 11 mm Hg on d 7. None of the manipulations of the reproductive organs caused detectable oxytocin release or increases in intramammary pressure, contrasting to responses to massage of the udder. Clitoral massage increased peak uterine pressure by 32 to 60% in four experiments. It did not induce luteinizing hormone release. The contraction was immediate, was not sustained and could be obtained repeatedly, suggesting a reflex response. Treatment with PGF2 alpha increased intramammary pressure. It increased uterine pressure on d 7, but had no effect at estrus. This contrasts with injected oxytocin, which resulted in the highest amplitude for cows in estrus, although the response on d 7 was greater in proportion to the low activity in controls at that time. It is concluded that manual manipulation of the reproductive tract (other than clitoral massage) has little effect on uterine contractility.
Animals↗
Normal and abnormal generation of isotype diversity.
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B-Lymphocytes↗
Differential recognition patterns of human immunoglobulin classes to antigens of Onchocerca gibsoni.
Adult Onchocerca gibsoni worms were fractionated into a surface-enriched fraction, a saline extract, a saline insoluble-detergent soluble fraction and a total glycoproteins extract. The antigens in each fractions were separated by electrophoresis in polyacrylamide gels and examined with an immuno-blot technique for reactive antibodies in sera from individuals infected with a variety of filarial and non-filarial nematode worms. Radiolabelled monoclonal antibodies were used to determine the Ig heavy chain isotypes. A number of antigens were demonstrated in all of the extracts, with many antigens of each extract being unique. Although some Onchocerca antigens stimulated antibodies of all human immunoglobulin classes, the panel of antigens recognized by each Ig isotype was different. The IgE response was restricted and directed at antigens not recognized by antibodies to other nematode parasites. IgM and IgA responses tended to recognize many antigens, whilst IgG responses were directed at intermediate numbers of antigens. The control of isotype balance to individual parasite antigens is thus independently regulated. This survey provides a rational basis for the exploration of Onchocerca antigen-human antibody class systems with relevance for diagnosis, protection and pathology.
Animals↗
Analysis of IgG subclass production in cell cultures from IgA deficient patients and in normal controls as a function of age.
IgA deficient individuals may also have low serum levels of IgG subclasses, especially IgG2. In the present study we examined the development of plasma cells producing IgM, IgA or IgG, and the IgG1 and IgG2 subclasses, following lipopolysaccharide (LPS) and pokeweed mitogen (PWM) stimulation of mononuclear cells (MNC) from normal and IgA deficient individuals as a function of age. Studies of blood MNC from 38 normal donors (age range 2-44 years) revealed an age-related distribution pattern of mu, gamma, alpha, gamma 1 and gamma 2 plasma cells produced in mitogen-stimulated and control cultures. Decreased IgA responses to both LPS and PWM were consistently observed in cultures of MNC from all of the nine children with IgA deficiency. When compared with age-matched controls the IgG response was also diminished in PWM stimulated cultures, whereas the IgM responses were normal. The IgG deficit was due to reduced responses for the gamma 1 and gamma 2 subclasses, and was most pronounced for IgG2; IgG2 plasma cell differentiation was particularly depressed in LPS cultures. In contrast to normal adult cells, blood MNC from the nine children with IgA deficiency and age-matched controls (2-17 years) yielded more IgG1 than IgG2 plasma cells in both control and LPS cultures, while the pattern of response to PWM was similar in all groups (gamma 1 greater than gamma 2). A good concordance was found between the level of secreted Ig in the culture supernatants and the relative number of IgM or, IgG and IgA plasma cells identified by immunofluorescence staining of cytoplasmic immunoglobulins.
Adolescent↗
Severe congenital leukopenia (reticular dysgenesis). Immunologic and morphologic characterizations of leukocytes.
We report fatal reticular dysgenesis in a premature infant presenting with severely decreased blood and bone marrow granulocytes and lymphocytes, an absent thymic shadow by x-ray film, and generalized lymphoid hypoplasia. Immunologic and electron microscopic evaluation of his white blood cells demonstrated that, despite extremely low cell numbers, cells from all stages of both granulocytic and lymphocytic development were present. Immature bone marrow cells of both myeloid and lymphoid lineages were found in much greater proportions than were mature cells; pre-B cells outnumbered B cells by more than tenfold. Megakaryocytes and erythroid cells appeared to be present in normal numbers, and tritiated-thymidine incorporation by bone marrow nucleated cells was also normal, although it may have largely occurred in erythroblasts. These data suggest that the primary defect in reticular dysgenesis is not failure in initiation of stem cell differentiation along lymphoid and myelomonocytic lines but rather an, as yet, undefined abnormality that interferes with normal growth and maturation of immune cells committed to these differentiation pathways.
B-Lymphocytes↗
The generation and regulation of human T lymphocytes by Imuthiol. Evidence from an in vitro differentiation induction system.
In vitro long term induction of human peripheral blood lymphocytes by sodium diethyldithiocarbamate, DTC (Imuthiol) enhanced the expression of OKT and HLA-DR phenotypic determinants. A population of T-B-DR+ cells was recruited from the null cell population. The increase in OKT+ cells, but not the enhanced HLA-DR expression appeared to be macrophage dependent. The surface markers of NK cells, monocytes or the pokeweed mitogen-induced IgM production were not modified. Collectively, the findings indicate that Imuthiol is not a B cell inducer, but is specifically active on the T-cell population in the way it induces the maturation of null cells into OKT+ DR+ lymphocytes.
Antibodies, Monoclonal↗
A human B cell differentiation antigen selectively expressed on mature B cells identified by a monoclonal antibody (HB-2).
A monoclonal IgM antibody (HB-2), produced against a membrane antigen on the Raji, B cell line, reacted by indirect immunofluorescence with 2 to 40% of lymphoblasts from the B cell lines, Raji, Daudi, SN-1036, and SB but not with other types of cell lines, including pre-B, myeloid, melanoma, or T cells. HB-2 antibody reacted with 10 +/- 3% of normal blood mononuclear cells, and was unreactive with monocytes, granulocytes, platelets, or erythrocytes. Two-color immunofluorescence revealed that HB-2 antigen expression was confined to cells bearing surface Ig. An interesting finding was the fact that 25% of plasmablasts induced by pokeweed mitogen also expressed the HB-2 antigen. However, pre-B and plasma cells from normal bone marrow did not express the HB-2 antigen either on their membrane surface or in their cytoplasm. Analysis of 85 leukemias revealed that the HB-2 antigen was expressed on acute and chronic B cell leukemias and Burkitt's lymphomas, but not on malignancies of pre-B, T, myelocytic, or plasma cells. The results suggest that expression of the HB-2 antigen is confined to mature B cells.
Antibodies, Monoclonal↗
Immunoglobulin gene rearrangements and deletions in human Epstein-Barr virus-transformed cell lines producing different IgG and IgA subclasses.
During differentiation B lymphocytes may switch from the expression of surface IgM to the synthesis of IgG, IgA, or IgE isotypes by using a different heavy chain constant region (CH) gene. The molecular mechanisms by which switching occurs remain controversial. Rearrangements and deletions of CH genes 5' to the expressed gene have often been observed in the mouse and, more recently, in human cells that have switched isotypes. We have used human JH, C micro, C gamma, and C alpha probes to examine the extent of the deletions and rearrangements in clones of Epstein-Barr virus-transformed human cells that produce IgG1, IgG3, IgG4, or IgA1. Though deletions of CH genes 5' to the expressed CH gene were consistently observed, the rearrangement process appeared to be highly variable for the nonproductive CH gene locus: deletion or persistence of 5' CH genes, combinations of deletion and duplication of 5' genes, and deletions extending to 3' CH genes. Our results reveal an unexpected lack of specificity in the DNA deletions in cells that have undergone isotype switching.
B-Lymphocytes↗
Uterine contractions and fertility following clitoral massage of dairy cattle in estrus.
Lactating and nonlactating Holstein cows received 30 s of gentle clitoral massage to determine its effect on uterine contractility and oxytocin concentrations in blood. Clitoral stimulation caused an immediate single uterine contraction, and this could be repeated at 2.5-min intervals. Oxytocin concentrations did not change during intensive blood sampling following clitoral massage. Thus, the uterine response is interpreted as neurally mediated. Eighteen full-time inseminators were allotted to two groups, balanced so that each day half were inseminating control and half treated animals (5 s of clitoral massage following insemination). The 56-day nonreturns for 2090 controls were 74.3% versus 74.0% for 2049 animals receiving clitoral massage. Heifers were more fertile than cows (81.9% versus 70.6% nonreturns), but there was no indication that clitoral massage altered the nonreturn rate in either age group. Also, the time of day animals were inseminated did not affect nonreturn rate.
Animals↗
Comparison of the expression of IL 2 receptors by human T and B cells: induction by the polyclonal mitogens, phorbol myristate acetate, and anti-mu antibody.
It is well established that IL 2 plays an important role in the proliferative response of T cells. Activated B cells were also recently found to express IL 2 receptors. The present studies were designed to compare qualitative, quantitative, and functional aspects of IL 2 receptor expression by activated T and B cells. Phorbol myristate acetate (PMA)-activated human T and small resting B cells and enhanced the expression of HLA-DR, HLA-DC/DS, and transferrin receptors while reducing Leu-4 antigen expression by T cells and IgM and IgD expression on B cells. PMA induced both T and B cells to express functional IL 2 receptors before cellular proliferation. Immune interferon did not participate in this induction. The m.w. of the IL 2 receptors expressed by activated T and B cells was identical: 54,000 to 59,000. Several differences were noted in the expression of IL 2 receptors by activated T and B cells on stimulation with PMA; T cells expressed IL 2 receptors sooner than B cells and in higher density, and the enhanced proliferative response of T cells to IL 2 was more difficult to inhibit with antibody to IL 2 receptors. In addition, IL 2 enhanced the expression of transferrin receptors by activated T cells but did not have a similar effect on activated B cells. Small B cells from the blood could also be induced by a mitogenic monoclonal anti-IgM antibody to express functional IL 2 receptors. Relatively large B cells in fresh blood samples were found to express functional IL 2 receptors and were capable of a modest proliferative response to IL 2. The intensity of the IL 2 receptor expression and the proliferative response by large B cells were enhanced by PMA stimulation. The data suggest that IL 2 receptors may play an auxiliary role in the B cell proliferative response and that IL 2 may exert its effect at a late phase in the B cell activation process.
Antibodies, Monoclonal↗
Human B cell responsiveness to B cell growth factor after activation by phorbol ester and monoclonal anti-mu antibody.
The effect of phorbol ester on human B cell activation was examined. Picomolar to nanomolar concentrations of phorbol ester induced a high level of proliferation in small IgM-positive B cells isolated from peripheral blood by fluorescence-activated cell sorting. The addition of optimal doses of anti-mu antibody resulted in enhanced proliferation of phorbol ester-activated B cells. The addition of B cell growth factor (BCGF) to phorbol ester-activated B cells also resulted in a dose-dependent synergistic effect and maximal enhancement on day 3. BCGF activity could be absorbed with either phorbol ester- or anti-mu-activated B cells, but not with resting B cells, thus confirming the induction of functional BCGF receptor expression. Cell proliferation was not necessary for the induction of functional BCGF receptors. Phorbol ester was a more efficient inducer of BCGF receptor expression than was anti-mu antibody; gamma-interferon treatment had no effect. BCGF enhanced transferrin receptor expression by phorbol ester-activated B cells. The results suggest that phorbol ester-activated small B cells can be used to monitor BCGF activity, and this synergistic combination may be useful in establishing BCGF-dependent B cell clones in culture.
Absorption↗
A large subpopulation of lymphocytes with T helper phenotype (Leu-3/T4+) exhibits the property of binding to NK cell targets and granular lymphocyte morphology.
A discrete subpopulation of lymphocytes sharing several phenotypic characteristics with natural killer (NK) cells was identified within the circulating pool of human lymphocytes that bear the T helper marker Leu-3. This Leu-3+ subpopulation of cells formed cell conjugates with the NK target cell lines K562 and MOLT-4, but did not bind to mouse myeloma and hybridoma cell lines that are insensitive to NK cells. The Leu-3+ lymphocytes binding to NK cell targets contained cytoplasmic granules similar in ultrastructure and cytochemistry to those previously defined in granular lymphocytes with NK function, except that the granules in Leu-3+ cells were smaller and fewer in number. Unlike classical NK cells, however, the granular Leu-3+ cells did not kill the target cells to which they bound, even after treatment with interferon. The proportion of granular Leu-3+ cells with the capacity to bind to NK cell targets was approximately 7% at birth and increased to approximately 21% of the Leu-3+ cells in adults. These observations suggest the possibility of a lineal relationship between the granular Leu-3+ cells and granular Leu-3- cells with NK capability.
Acid Phosphatase↗
Expression of the Chediak-Higashi lysosomal abnormality in human peripheral blood lymphocyte subpopulations.
Fusion of lysosomes to form a giant cytoplasmic inclusion is a major abnormality expressed by multiple hematopoietic and non-hematopoietic cell types in Chediak-Higashi (C-H) patients. In this study, the extent of involvement of lymphoid cell subpopulations was defined. Purified populations of B cells, natural killer (NK) cells, and helper T cells were obtained from two C-H patients and normal controls by immunofluorescence staining of their blood mononuclear cells with the monoclonal antibodies HB-2, Leu-7, or Leu-3 followed by fluorescence-activated cell sorting. Cytochemical and ultrastructural analyses as well as functional assays were performed to determine whether or not the C-H lysosomal abnormality was expressed in the different lymphocyte subpopulations. B cells expressed the C-H defect following activation and differentiation. All of the Leu-7+ cells and a significant proportion of the Leu-3+ cells displayed the C-H abnormality. These Leu-3+ cells share the NK lineage characteristics of granular lymphocyte morphology and the capacity to bind to NK cell targets. In contrast, the C-H abnormality was not observed in non-NK target-binding cells with T helper phenotype, in which clusters of lysosomes formed a normal Gall body. Moreover, T cell functions were unimpaired in C-H patients. These observations raise the issue of the lineal relationship between granular and nongranular lymphocytes typed as T cells on the basis of cell surface antigen markers.
Adult↗