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Biomedical subjects

A D Bankhurst

Publications and source records attributed to A D Bankhurst.

At least 55 records · Page 3Linked to original sources

Family distribution of anti-F(ab')2 antibodies in relatives of patients with systemic lupus erythematosus.

Recently we reported an inverse relationship between the levels of anti-F(ab')2 antibodies and disease activity in systemic lupus erythematosus (SLE). The present study focused on anti-F(ab')2 antibodies in unaffected relatives of SLE patients. Sixty sera from first degree family members from 11 SLE families and 49 sera from 8 control families were studied. Percentage of SLE family members with anti-DNA antibodies (15%) was higher than than control family sera (8%, P less than 0.05). Anti-F(ab')2 antibodies were measured using ELISA assays. The SLE family sera had higher amounts of anti-F(ab')2 antibodies than the normal control family group (P = 0.0051). In an effort to determine if anti-F(ab')2 antibodies found in high titres in the sera of some SLE family members had specificity for the F(ab')2 fragment of anti-DNA antibodies of the SLE relative patients, DNA-anti-DNA inhibition experiments were performed using anti-F(ab')2 prepared from the relative in parallel with anti-F(ab')2 prepared from normal controls with equivalent high titres of serum anti-F(ab')2. Inhibition exhibited by anti-F(ab')2 of first degree relatives was higher than that obtained from control normal donors (P less than 0.02). Such differences in inhibition were not recorded using a control tetanus toxoid-anti-tetanus toxoid assay. In direct binding ELISA experiments, peroxidase-conjugated anti-F(ab')2 antibodies from the same first degree relative showed high relative specificity against purified anti-DNA antibodies of his SLE proband when compared to those obtained against different anti-DNA antibodies isolated from unrelated SLE patients (P less than 0.001). Such a substantial difference was not observed in parallel experiments using peroxidase conjugated anti-F(ab')2 antibodies from normal controls unrelated to SLE subjects.

Adolescent↗

Natural killer cells in connective tissue disorders.

NK cells may be important in the elimination of cells infected by virus, in the regulation of antibody production, and in tissue destruction. The significance of NK cells in rheumatic disorders is unknown, but NK cells and NK-like cells have been found in the peripheral blood and synovial tissues of patients with autoimmune disease. In particular, defects in NK cell activity have been reported in SLE, RA, PSS and SS. Of these diseases, SLE appears to be the best characterized with obvious abnormalities in NK cell numbers, impaired cytotoxicity of individual NK cells, decreased release of cytotoxic factors, deranged IFN modulation of the NK cell, and associated abnormalities in the IL-2 system. The association of these abnormalities with the underlying disease process is currently under investigation.

Arthritis, Rheumatoid↗

The effect of beta-endorphin on natural cytotoxicity and antibody dependent cellular cytotoxicity.

The ability of the central nervous system to modulate immune responsiveness has received increasing attention. A potential mechanism that would allow the central nervous system to alter the immune system is the release of neuroendocrine and neurotransmitter polypeptides into the peripheral circulation with subsequent modulation of immunocyte function. In this report, we demonstrate that the neuropeptide, beta-[D-ALA2]-endorphin augments natural cytotoxicity but does not effect antibody-dependent cellular cytotoxicity. The observations are discussed in relation to the mechanisms for natural cytotoxicity and antibody dependent cellular cytotoxicity.

Antibody-Dependent Cell Cytotoxicity↗

Impaired release of a soluble natural killer cytotoxic factor in systemic lupus erythematosus.

Disease states characterized by abnormalities in immune regulation often demonstrate concomitant abnormalities in cytotoxicity mediated by natural killer (NK) cells. For example, some patients with systemic lupus erythematosus (SLE) have depressed NK activity despite the presence of normal numbers of effector cell:target cell conjugates. This study was designed to determine if defects in NK cell function were directly related to impaired release of a soluble cytotoxic factor. NK activity of peripheral blood mononuclear cells and large granular lymphocytes was measured using 51Cr-labeled K562 target cells in 4-hour release assays. The SLE patients had significantly decreased NK activity relative to normal controls. However, the number of effector cell:target cell conjugates was not different in SLE patients versus control subjects. The release of a soluble natural killer cytotoxic factor (NKCF) by peripheral blood mononuclear cells was measured by cytotoxicity induced in K562 cells. NKCF was released preferentially by suspensions enriched in NK cells (large granular lymphocytes). At a 1:1 dilution, NKCF release was significantly lower in SLE patients than in controls. The release of NKCF correlated well with NK activity. Thus, this study shows that the defect in NK cell activity in SLE patients may be related to an impairment in release of a soluble cytotoxic factor with specificity for NK cell-sensitive targets.

Adult↗

Studies of anti-F(ab')2 antibodies and possible immunologic control mechanisms in systemic lupus erythematosus.

Studies of anti-DNA and anti-F(ab')2 antibodies were performed by enzyme-linked immunosorbent assay in 51 patients with systemic lupus erythematosus (SLE). Patients with severe, uncontrolled disease showed high levels of anti-DNA and low levels of anti-F(ab')2 antibodies. Patients with quiescent SLE usually showed high levels of anti-F(ab')2 and low levels of anti-DNA antibodies. Isolated anti-F(ab')2 antibodies from autologous SLE remission serum or from the sera of unaffected siblings of SLE patients showed maximum inhibition in test systems using affinity-purified SLE anti-DNA antibodies reacting with single-stranded DNA.

Adult↗

Interferon-alpha regulation of lymphocyte function in systemic lupus erythematosus.

Interferon (IFN) production and responsiveness are abnormal in some patients with systemic lupus erythematosus (SLE). The present investigation was designed to further delineate these abnormalities of IFN response. The response to IFN-alpha was determined in peripheral blood mononuclear cells (PBMC) using the natural killer (NK) cell assay, the concanavalin A (Con A)-induced T-cell proliferation assay, and the pokeweed mitogen (PWM) blastogensis system. The average NK cell activity was impaired in SLE patients (19.7 +/- 3.8 LU/10(7) cells; n = 22) compared to 24 normal controls (45.6 +/- 7.2 LU/10(7) cells; n = 24) (P less than 0.05). In addition, the response to IFN in the NK cell system was impaired in SLE (110.0 +/- 38.0% enhancement) compared to controls (320.0 +/- 94.0% enhancement; +/- SD P less than 0.05). In contrast to the impaired IFN response of the NK cell in SLE, the effect of IFN on Con A- and PWM-induced blastogenesis in SLE was normal (P greater than 0.1). Thus, the defect in IFN response in SLE appeared to be confined to the NK cell system, but was not present in the other assay systems. To determine if the impaired IFN response might be secondary to impaired release of natural killer cytotoxic factor (NKCF) induced by IFN, NKCF was generated from PBMC in the presence of IFN. IFN-induced NKCF release was markedly impaired in SLE (8.9 +/- 11.7%) relative to normal controls (22.3 +/- 11.2%) (P less than 0.05). IFN-induced release of NKCF in SLE also closely correlated with IFN-induced NK enhancement (r = .83, P less than 0.05). No depletion of NK cells was noted using the monoclonal antibody HNK-1. Thus, this study demonstrates for the first time that the insensitivity to IFN in SLE is not a universal lymphocyte defect but appears to be isolated to the NK cell which is functionally abnormal.

Adult↗

Lymphokines in autoimmunity: relationship between interleukin-2 and interferon-gamma production in systemic lupus erythematosus.

Interleukin-2 (IL-2) and interferon (IFN) are potent immunoregulatory factors. Recently, it has been demonstrated that IL-2 production is necessary for IFN-gamma synthesis. Thus, defects in IL-2 production could lead to defects in IFN-gamma production. Indeed, in systemic lupus erythematosus, defects in IFN-gamma and IL-2 production have been noted. To test this hypothesis, IL-2 and IFN-gamma production rates by peripheral blood mononuclear cells PBMC were measured simultaneously after stimulation by concanavalin A (Con A) and phorbol myristic acetate (PMA). IL-2 activity was determined employing HT-2, a cell line with an absolute growth requirement for IL-2. IFN-gamma activity was assessed using a standard viral plaque inhibition assay. IL-2 production in normal controls (n = 6) was 14.7 +/- 9.4 units/ml and in SLE patients (n = 10), 18.0 +/- 10.5 units/ml (P greater than 0.05). IFN-gamma production in controls was 74.7 +/- 43.7 units/ml and in SLE patients, 68.8 +/- 35.4 units/ml (P greater than 0.05). Only one SLE patient, who had moderately active disease, demonstrated defects in IL-2 synthesis (less than 4 units/ml) and IFN-gamma production (16 units/ml). Thus, production of these lymphokines in SLE patients was largely normal in response to Con A-PMA. These results imply that the intrinsic pathways of IFN and IL-2 production are basically normal in most SLE patients although defects in production of these lymphokines can occur. The defects in IL-2 and IFN production in SLE previously reported may be secondary to an impaired cellular response to the selected inducing agent rather than a primary defect in the actual ability to produce these important lymphokines.

Autoantibodies↗

Defects in natural killer cell activity and interferon response in human lung carcinoma and malignant melanoma.

The natural killer (NK) activity of peripheral blood mononuclear cells from 25 patients with squamous cell carcinoma of the lung, malignant melanoma, or epitheloid cancers of the gastrointestinal tract was measured by the lysis of 51Cr-labeled K562 target cells. NK activities of many patients with lung cancer or malignant melanoma were decreased relative to normal controls. This abnormality was significantly correlated with advancing stage of disease and the percentage of monocytes in the cell suspensions. Addition of indomethacin or removal of monocytes did not restore depressed NK function to normal levels. Abnormalities of NK function did not appear to be secondary to the presence of mononuclear suppressor cells. The response to interferon was also impaired in patients with advanced disease. The number of effector:target conjugates was normal even in patients with depressed NK function; however, the number of active lytic effectors was decreased. These results imply that the cells which bind tumor targets are present in patients with advanced cancers, but these cells are either immature or functionally inactive.

Humans↗

Separation of natural and antibody-dependent cytotoxicity by differential reactivity of human mononuclear cell Fc receptors with IgG.

Nylon wood non-adherent, human peripheral blood mononuclear cells which are reactive with the OKM1 monoclonal antibody could be separated into two subpopulations based on their Fc receptor reactivity with human monomeric IgG (FcR-IgG) using flow cytometry. The majority of natural killer cells was found primarily in the OKM1+ subset with low FcR reactivity. In contrast, effector cells for antibody-dependent cytotoxicity (K cells) were found in subsets with both high and low FcR-IgG reactivity.

Adult↗

Exophthalmos in systemic lupus erythematosus.

In systemic lupus erythematosus (SLE) exophthalmos is distinctly rare. This article describes the first patients with SLE in whom exophthalmos was preceded by pseudotumor cerebri. The apparent lack of resolution of the exophthalmos with corticosteroids in our case emphasizes the potential need for therapeutic alternatives such as immunosuppressive agents, orbital irradiation, and/or plasmapheresis.

Adult↗

Enrichment of natural killer cells by negative selection: comparison to Percoll gradient separation method.

A method is described for the preparation of human peripheral blood mononuclear cell (PBMC) suspensions containing highly enriched natural killer (NK) cell cytotoxicity. The technique involved the negative selection of OKM1+ cells by the selective removal of nylon wool nonadherent PBMC which are reactive with the Leu-1 monoclonal antibody. The Leu-1+ cells are removed by subsequent rosette formation with anti-mouse IgG coated bovine erythrocytes. The resultant OKM1+ cell suspension had a greater number of large granular lymphocytes, K562 target binding effector cells, and lytic activity than concomitantly prepared fractions of Percoll gradients.

Antibodies, Monoclonal↗

Human alpha-interferon enhances in vitro IgM rheumatoid factor synthesis by lymphocytes from normal subjects and rheumatoid arthritis patients.

Alpha-interferon (alpha-IFN) was added to pokeweed mitogen (PWM) or Epstein-Barr virus stimulated human peripheral blood mononuclear cell cultures from normal subjects or patients with rheumatoid arthritis (RA). Alpha-IFN enhanced in vitro production of PWM induced IgG and IgM, and significantly enhanced PWM induced IgM rheumatoid factor (IgM-RF) production by lymphocytes both from normal subjects and RA patients. Enhancement was recorded whether cells were preincubated with alpha-IFN for 16 hours or with alpha-IFN present throughout the culture period. Alpha-IFN did not enhance IgM-RF production in the absence of PWM or T cells. Enhancement of IgM-RF production was not seen in Epstein-Barr virus stimulated cultures.

Adult↗

Impaired natural killer cell function in systemic lupus erythematosus. Relationship to interleukin-2 production.

Immune regulation requires clonal expansion of regulatory T cells which is dependent on the presence of interleukin-2 (IL-2). Previously, both natural killer (NK) cell function and IL-2 production were found to be depressed in systemic lupus erythematosus (SLE). This study was designed to determine the relationship between IL-2 production and NK cell activity in SLE. NK activity as determined by a standard 4-hour 51Cr-release assay was impaired in SLE (11.0 +/- 5.1 lytic units/10(7) cells) compared with controls (25.1 +/- 7.1 lytic units/10(7) cells) (P less than 0.05). IL-2 production was induced with concanavalin A and phorbol ester and quantitated using the IL-2 dependent cell line HT-2. IL-2 production was impaired in only 1 SLE patient, despite concomitant abnormalities in NK function in the SLE group as a whole. Moreover, in patients with impaired NK activity, incubation of lymphocytes with exogenous IL-2 did not restore NK activity to normal levels. These findings demonstrate that impaired NK activity in SLE is independent of IL-2 production and that defects in IL-2 production in SLE may not be as common as previously reported.

Humans↗

Abnormal interferon modulation of natural cytotoxicity in systemic lupus erythematosus. Relation to suppressive serum factors.

In the present study we examined one aspect of the derangement in natural cytotoxicity (natural killer, NK) activity observed in some patients with systemic lupus erythematosus (SLE), i.e., the lack of enhancement of NK activity usually seen with interferon (IFN). NK activity of SLE patients as a group was 23.0 +/- 3.9 lytic units (LU)/10(7) cells (mean +/- SE). This contrasted with the NK activity found in normal controls (45.0 +/- 3.8 LU/10(7) cells) (P less than 0.05). The enhancement seen with IFN was an increase of 15.4 +/- 4.0 LU/10(7) cells in SLE patients compared with 104.6 +/- 192 LU/10(7) cells in control subjects (P less than 0.05). SLE sera and aggregated IgG (Agg-IgG) also inhibited the increase in NK activity of normal peripheral blood mononuclear cells after IFN priming. The results reported here support the hypothesis that the impaired baseline NK activity and poor response to IFN noted in SLE are secondary, in part, to the presence of inhibitory serum factors and preactivation by IFN.

Cells, Cultured↗

Natural killing and antibody-dependent cellular cytotoxicity: characterization of effector cells by E-rosetting and monoclonal antibodies.

Recent investigations have indicated that the OKM1 hybridoma monoclonal antibody reactive with cells of the myelomonocytic series identifies a subpopulation of human peripheral blood mononuclear cells (PBMNC) which mediate natural and antibody-dependent cellular cytotoxicity (ADCC). However, it was not clear whether this OKM1+ group was heterogeneous with regard to cytotoxic function or the presence of receptors for sheep erythrocytes. Thus, the purpose of the present study was to further define the phenotype of the ADCC effector cell and natural killer (NK) cell using a combination of reactivity with hybridoma antibodies and separation of subsets by sheep erythrocyte rosette (E+) formation. Furthermore, the phenotypes of the NK population were assessed directly by performing two-color immunofluorescent staining on tumor cell conjugates. These studies led to the following conclusions: (1) that NK activity is mediated by both E+ OKM1+ and E- OKM1+ cells; the E+ OKT3+ cell possessed essentially no ADCC or NK activity; (2) that E+ OKM1+ cells mediated more NK activity on a per cell basis than E- OKM1+ cells; this was verified by separating OKM1+ cells on a cell sorter into E+ and E- with the OKT11 monoclonal antibody (anti-E-receptor antibody); (3) that E+ OKM1+ cells mediated both ADCC and NK activity; (4) that the phenotypes of PBMNC forming tumor cell conjugates were (a) OKM1+ (both E-receptor positive and negative) and (b) OKM1- E-receptor positive.

Antibodies, Monoclonal↗

Characterization of the suppressor activity in lymphocytes from patients with common variable hypogammaglobulinemia: evidence for an associated primary B-cell defect.

The pathogenetic mechanisms responsible for the impaired immunoglobulin production in common variable hypogammaglobulinemia (CVH) are diverse with abnormalities in both B cells and immunoregulatory T cells. Production of IgG, IgM, and IgM-rheumatoid factor (IgM-RF) was measured in pokeweed mitogen (PWM) or Epstein-Barr virus (EBV)-stimulated cultures using various combinations of CVH, cord blood mononuclear cells (CBMC), and normal adult control B and T cells. The following results were obtained. First, the proportion of OKT3+ and OKT8+ cells were increased in CVH patients. Second, the T cells from four CVH patients and CBMC suppressed PWM-induced IgG, IgM, and IgM-RF production by normal B cells. Furthermore, major suppressor activity was found in the OKT8+ T-cell subpopulations in CBMC and three out of four CVH patients. There was no significant difference in relative suppression by OKT8+ cells from normal adults, CVH patients, or CBMC. However, in one CVH patient suppressor T cells were found in both OKT4+ as well as OKT8+ fractions. In the CVH patient with OKT4+ suppressor cells, X irradiation (1250 rads) abrogated suppressor activity and restored helper activity in the OKT4+ T-cell fraction. Irradiation of normal OKT4+ cells did not increase helper activity. When non-E-rosetting cells from normal subjects, CVH, and CBMC were stimulated with EBV it was observed that normal adult B cells could be induced to secrete IgG, IgM, and Ig-RF whereas CVH and CBMC could only produce IgM and IgM-RF but not IgG. The present study demonstrates for the first time that a radiosensitive OKT4+ suppressor cell is present in some CVH patients.

Agammaglobulinemia↗

Studies on the relationship between HLA DR4 and in vitro IgM rheumatoid factor production.

The relationship between the presence of the DR4 antigen and other HLA antigens and in vitro production of IgM-rheumatoid factor by lymphocytes from a group of healthy young subjects was examined. Pokeweed and Epstein--Barr virus-stimulated lymphocyte cultures were examined for the production of rheumatoid factor and immunoglobulins. No significant correlation was found between the presence of the DR4 antigen and in vitro production of IgM rheumatoid factor. The presence of the B18 antigen seemed to identify a population of nonresponders when stimulated with PWM.

Adult↗

Natural killer cell in systemic lupus erythematosus. Defects in effector lytic activity and response to interferon and interferon inducers.

Spontaneous cytotoxicity mediated by natural killer (NK) cells is impaired in several human diseases including systemic lupus erythematosus (SLE). The precise mechanism(s) by which NK activity is suppressed in patients with SLE is generally unknown. The present study was designed to focus on cellular defects per se in NK cells from patients with SLE. It was observed that the usual enhancing effect of interferon (IF) and IF inducers was markedly impaired in SLE patients. Of 24 SLE patients studied, 17 had significantly decreased NK activity relative to controls. NK activity had a significant negative correlation with clinical activity score (r = -0.56, P less than 0.005) but was not correlated with corticosteroid dose, antinuclear antibody titers, total hemolytic complement (CH50), or sedimentation rate. Furthermore, significant depressions in NK activity correlated with variations in disease activity in six patients followed serially. Depressed NK function could not be reversed by prolonged in vitro incubation at 37 degrees C or with protease treatment. Furthermore, depressed NK activity was not altered by removal of glass adherent cells nor was a suppression of NK activity in normal controls seen by the addition of SLE peripheral mononuclear cells. No reversal of depressed activity to normal levels was seen by the addition of indomethacin nor did the supernatants from SLE cell cultures cause a suppression of normal NK function. NK activity in SLE patients did not respond normally to IF inducers (poly-I:C and concanavalin A) even if the SLE patients had normal NK function. The response of SLE cells to exogenous IF was also impaired. The number of effector-target conjugates was quantitated with several target cells (K562, Yac-1, Fravel) in SLE patients and controls. A significant correlation between the proportion of glass nonadherent mononuclear cells that formed effector-target conjugates with these various targets and the magnitude of NK lysis was observed. However, SLE and normal subjects had equal numbers of effector-target conjugates independent of NK function. Release of a soluble cytotoxic factor was induced with concanavalin A, and was markedly impaired in SLE patients relative to normal controls. Thus, impaired NK cell function in SLE does not appear to be related to cell-mediated suppressive mechanisms or to the deletion of effector cells; rather, the decreased NK activity may be related to an impaired release of a soluble cytotoxic factor.

Adult↗