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Biomedical subjects

B Golding

Publications and source records attributed to B Golding.

At least 73 records · Page 4Linked to original sources
PubMed1987

In vitro generated human monoclonal trinitrophenyl-specific B cell lines. Evidence that human and murine anti-trinitrophenyl monoclonal antibodies cross-react with Escherichia coli beta-galactosidase.

Stable human antigen-specific monoclonal B cell lines were established without prior in vivo immunization. This was accomplished by expanding the anti-trinitrophenyl (TNP) B cells in vitro with the antigen TNP-Brucella abortus and then immortalizing them with Epstein-Barr virus. Five anti-TNP clones were selected by sequential limiting dilution. All five anti-TNP clones secreted IgM kappa antibodies. When tested against a panel of self and environmental antigens, all five anti-TNP clones exhibited cross-reactivity with an Escherichia coli-derived beta-galactosidase. To determine whether this was a more general phenomenon, a panel of murine monoclonals were tested and found to bind to beta-galactosidase. It is therefore possible that human and murine anti-TNP beta cell responses reflect reactivity against an environmental antigen, namely an epitope present on E. coli-derived beta-galactosidase. This approach of expanding human antigen-specific B cells by antigen stimulation in vitro, with a T-independent hapten-carrier conjugate before Epstein-Barr virus transformation, may prove useful in the development of human monoclonals for therapeutic purposes.

Animals↗
PubMed1986

The role of nonactivated and interferon-gamma activated monocytes in regulating normal and SLE patient B cell responses to TNP-Brucella abortus.

We have previously characterized the human B cell response to trinitrophenol (TNP)-Brucella abortus (Ba) response as being T cell independent. In this report we examine the role of monocytes in the TNP-Ba antibody response of human peripheral blood mononuclear cells (PBMC). Depletion of monocytes by sequential adherence to plastic and Sephadex G-10 passage did not result in decreased plaque-forming cell responses to TNP-Ba, suggesting that monocytes were not required. On the contrary monocytes were probably inhibitory because their removal resulted in enhanced responses. This was confirmed by showing that adding monocytes back reconstituted the inhibition. When interferon-gamma (IFN-gamma), a potent activator of monocytes, was added to TNP-Ba-driven PBMC cultures, marked inhibition (greater than 90%) of the responses ensued. This IFN-gamma-mediated suppression was monocyte dependent because it was completely abrogated by monocyte, but not T cell depletion. Previously, we described a concanavalin A (Con A), T cell inhibition pathway of the TNP-Ba response. Both the Con A and IFN-gamma pathways were tested for their ability to inhibit systemic lupus erythematosus (SLE) patient responses to TNP-Ba. The B cell response of SLE patients was inhibitable by both pathways. In all of the patients, the inhibition was complete (greater than 95%) when IFN-gamma was added to the cultures. In the presence of Con A, greater than 95% inhibition was observed in six of 10 patients, the remainder being inhibited to a lesser extent. Thus the hyperactive B cells from SLE patients can be down-regulated, particularly in the presence of IFN-gamma.

B-Lymphocytes↗
PubMed1984

Human thymus-independent antibody response in vitro: III. Precursor frequencies and fine specificity of human B cells stimulated by TNP-Ba in the presence and absence of Con A.

Previously, we reported that human B lymphocytes can be stimulated by trinitrophenylated Brucella abortus (TNP-Ba) in vitro to generate T-independent (TI) hapten specific plaque-forming cells (PFC). Furthermore we showed that addition of Con A on Day 3 of culture enhanced the anti-TNP response. In this report the characteristics of the anti-TNP PFC responses elicited by TNP-Ba in the presence or absence of Con A were further defined by estimating precursor frequencies and clone sizes, and by assessing the diversity of the anti-TNP-PFC response using hapten inhibition profiles. Addition of Con A to TNP-Ba-stimulated cultures was associated with a 2.3- to 3.6-fold increase in anti-TNP precursors and was also accompanied by a more vigorous clonal expansion. Fine specificity analysis of anti-TNP PFC revealed that Con A addition resulted in PFC with unique hapten inhibition profiles in that they were less inhibitable by TNP-EACA, TNP-lysine, and DNP-lysine but more inhibitable by DNP-glycine when compared to PFC generated by TNP-Ba alone. These findings suggest that at least some of the additional precursors recruited in the presence of Con A are qualitatively distinct from those activated by TNP-Ba alone since they express different V region gene products.

Antibodies, Bacterial↗
PubMed1984

Newborn and Wiskott-Aldrich patient B cells can be activated by TNP-Brucella abortus: evidence that TNP-Brucella abortus behaves as a T-independent type 1 antigen in humans.

TNP-Brucella abortus (TNP-Ba) has been classified as a T-independent type 1 (TI-1) antigen in the mouse on the basis that it activates neonatal and CBA/N (X-linked immunodeficient) murine B cells in contrast to T-independent type 2 (TI-2) antigens. Therefore, it was of interest to determine whether human newborn and X-linked Wiskott-Aldrich syndrome B cells could be triggered by TNP-Ba. Previous studies had shown that human B cells from both these latter sources were relatively insensitive to stimulation with T-dependent and polysaccharide antigens (TI-2 in mouse). In this study, we show that TNP-Ba can trigger human cord blood B cells to differentiate into anti-TNP plaque-forming cells (PFC) in a hapten-specific and T-independent manner. The dose response and kinetics were similar to those previously seen with adult cells. The newborn responses, however, were lower than adult PFC responses. Precursor frequency and clone size analyses revealed that this lower response was not due to newborn cells containing fewer precursors but was the result of a reduced ability of these anti-TNP clones to expand. The ability of TNP-Ba to activate immature newborn B cells implies that this antigen can be used to assess B cell function in very young children. It also implies that TNP-Ba behaves as a TI-1 antigen in humans as well as in mice. This was supported by the finding that B cells from Wiskott-Aldrich patients, which were unreactive to polysaccharide antigens, were generally responsive to TNP-Ba. Therefore, it would appear that human newborn and Wiskott-Aldrich patients do possess a functionally competent B cell subset possibly equivalent to Lyb-5- immature murine B cells.

Antibodies, Bacterial↗
PubMed1983

CBA/N X-linked defect delays expression of the Y-linked accelerated autoimmune disease in BXSB mice.

BXSB male mice spontaneously develop progressive autoimmune disease characterized by high serum immunoglobulins, including anti-nuclear antibodies (ANA), enlarged spleen and lymph nodes, and diffuse proliferative glomerulonephritis. Females develop symptoms at a much slower rate. The mechanisms underlying the autoimmune disease and the nature of the Y-linked accelerating factor have not yet been elucidated. We found that the male progeny of the cross between the non-autoimmune strain CBA/Ca and BXSB (CBA/Ca X BXSB)F1 showed progressing signs of autoimmunity starting at 6 to 7 mo. In contrast, the male progeny that resulted from BXSB males crossed with immune-defective CBA/N females (Xid) were devoid of splenic B colonies, were nonresponsive to TNP-Ficoll, and were free of autoimmune disease for at least 10 mo. At 18 mo, some of the (CBA/N X BXSB)F1 mice developed weak antinuclear antibodies, but no spleen or lymph node enlargement was seen. The same mice had low anti-TNP Ficoll responses but did not produce B colonies in vitro. The role of the X chromosome in regulating expression of autoimmunity in young and old BXSB mice is discussed.

Aging↗
PubMed1982

Lectin-induced regulation of human thymus-independent anti-TNP responses in vitro.

Recently we showed that human hapten-specific T cell-independent (TI) antibody responses could be elicited in vitro with the antigen trinitrophenylated Brucella abortus (TNP-Ba). Although by definition T cells are not required for TI responses, they have also shown capable of regulating such responses in the mouse. In this study we examine the ability of the T cell lectins Con A and PHA to modulate TI responses of human tonsil cells. Addition of the lectins to cultures on day 0 or day 3 resulted in inhibition (greater than 90%) or enhancement (greater than 150%) of the anti-TNP PFC response, respectively. The inhibition was only apparent if the lectins were added at the onset of culture but not 24 hr later, and was T cell-dependent, because T cell-depleted cultures (less than 0.5% E-rosetting cells) could not be inhibited. The enhancement observed was even greater in T cell-depleted cultures and was antigen-specific because TNP-SRBC but not PC-SRBC targets were lysed and the PFC were inhibited by soluble TNP (greater than 70%). This finding suggested that anti-TNP PFC precursors were proliferating more vigorously or that additional, previously antigen-insensitive cells were being recruited in the presence of lectin.

Antibody Formation↗