Lymphocyte Immunoglobulin Research Articles

Differential effect of opioids on immunoglobulin production by lymphocytes isolated from Peyer's patches and Spleen

The mucosal immune system plays an important role in blocking the penetration of invasive organisms into various mucosal surfaces. Evidence now suggests neuroendocrine peptide hormones have immunomodulatory properties, including the ability to alter mucosal immunity. The potential for opioid compounds and corticotropic hormone (ACTH) to modulate mucosal immune function was investigated. We have found β-endorphin, ACTH, and naltrindole (delta-class opioid receptor antagonist) to significantly suppress concanavalin A-stimulated Peyer's patch lymphocyte immunoglobulin production of IgA, IgG, and IgM isotypes. Oxymorphindole, a delta class opioid receptor agonist, significantly decreased IgM but not IgA or IgG production by the mitogen-stimulated Peyer's patch lymphocytes. Both oxymorphindole and naltrindole modestly reduced interleukin-2 receptor expression of concanavalin A- (Con A)-stimulated splenic and Peyer's patch lymphocytes. Neither compound appreciably affected immunoglobulin production by lipopolysaccharide-stimulated Peyer's patch lymphocytes. Collectively, these results indicate stress-related peptides such as ACTH and opioids may be involved in the regulation of immunoglobulin synthesis by Peyer's patch lymphocytes.

Exogenous interleukin-2 does not reverse the immunoinhibitory effects of 1,25-dihydroxyvitamin D 3 on human peripheral blood lymphocyte immunoglobulin production

1,25-Dihydroxyvitamin D 3 (1,25-D 3) is known to have potent inhibitory effects on human peripheral blood mononuclear cell (PBMC) functions. Previous experiments suggest that addition of interleukin-2 (IL-2) to cell cultures can reverse the antiproliferative action of 1,25-D 3. Previous studies have also shown that the CD4 + T-cell subset is more sensitive to the antiproliferative actions of 1,25-D 3 than are the CD8 + T-cells. The objective of this study was to determine whether exogenous IL-2 could reverse the antiproliferative and immunoinhibitory action (inhibition of Ig production) in mitogen-activated PBMC cultures and in fluorescein-activated cell sorting (FACS) experiments where CD8 + T-cells were removed from PBMCs before mitogen stimulation with/without exogenous IL-2 added. In these studies, addition of IL-2 to mitogen-activated, 1,25-D 3-treated PBMCs allowed the cells to overcome the 1,25-D 3 suppressive effect on cell proliferation. However, exogenous IL-2 did not overcome the 1,25-D 3-mediated inhibitory effect on PBMC Ig production. Using FACS lymphocyte populations (CD4 +, CD8 + and B-cells), we showed that CD4 + T-cell-directed Ig synthesis in co-culture with autologous B-cells was inhibitable by incubation of cells with 1,25-D 3, but Ig synthesis was restored to near-normal levels by addition of exogenous IL-2. This clearly contrasts with the inability of IL-2 to reverse the 1,25-D 3 inhibitory effect on Ig synthesis in PBMCs. In other experiments, when CD8 + cells were removed from mitogen-stimulated, 1,25-D 3-treated PBMCs, addition of exogenous IL-2 resulted in a full reversal of the 1,25-D 3-mediated Ig inhibition. These data suggest that the inability of IL-2 to reverse the inhibitory effects of 1,25-D 3 on PBMC Ig production is probably a result of a lack of sensitivity of CD8 + T-cells to the antiproliferative and immunoregulatory actions of 1,25-D 3. This is possibly because of a differential expression of 1,25-D 3 receptors on CD 4+ and CD8 + T-cells.

Immunoglobulin (Fc) receptors on murine T- and B-lymphocytes: Investigations using tumor models

Lymphoid tumors are productive experimental models for the study of lymphocyte immunoglobulin receptors. Investigations with Fc receptor expressing lymphoid tumor cells have generated much useful information about: (a) the developmental expression of the different classes of Fc receptors on lymphoid cells of the T- and B-lineages; (b) the biochemical steps involved in the regulation of Fc receptor expression on lymphoid cells; (c) the structures of lymphoid cell Fc receptors and their genes; (d) the signals that induce alterations in the expression of Fc receptors on lymphoid cells; and (e) the molecular specificity of the binding of immunoglobulin to lymphoid cells Fc receptors. In addition, tumors that secrete immunoglobulins are providing useful models for analysis of the mechanisms by which B-cells influence Fc receptor expression and function on T-cells. An interesting, bi-directional immunoregulatory circuit involving Fc epsilon R+ host T-cells and IgE-secreting hybridoma cells has been identified that could prove useful in the analysis of the regulation of epsilon heavy chain expression. The studies discussed in this article and elsewhere in this volume serve to emphasize that, in addition to being clonal sources of key molecules such as Fc receptors and their messenger RNAs, lymphoid tumor cells that express Fc receptors are powerful and unique experimental models for investigating the developmental biology, regulation and function of lymphocyte Fc receptors.

Divergent effects of silica on lymphocyte proliferation and immunoglobulin production.

Silicosis is an interstitial lung disorder that is frequently associated with hypergammaglobulinemia and increased numbers of lymphocytes at sites of disease. To determine the effect of silica on the generation of immunoglobulin-secreting cells, mononuclear cells were stimulated with antigens or mitogens and placed into 1) high-density cultures (2.5 X 10(6) cells/ml) that were not exposed to silica, in which pokeweed mitogen (PWM)-induced generation of immunoglobulin-secreting cells was suppressed by the presence of monocytes; or 2) low-density cultures (0.5 X 10(6) cells/ml) that were not exposed to silica, in which PWM-induced generation of immunoglobulin-secreting cells was not suppressed. Silica added to PWM-stimulated high-density cultures significantly increased the numbers of immunoglobulin-secreting cells. Silica also significantly increased the numbers of immunoglobulin-secreting cells in high-density cultures stimulated with purified protein derivative and tetanus toxoid and augmented the proliferation of phytohemagglutinin-stimulated mononuclear cells (P less than 0.05). In contrast to high-density cultures, silica added with PWM to low-density cultures reduced the numbers of immunoglobulin-secreting cells. These studies suggest that silica can have potent regulatory effects on various cellular immune processes that are relevant to the lung.

Influence of ofloxacin and pefloxacin on human lymphocyte immunoglobulin secretion and on polymorphonuclear leucocyte superoxide anion production.

The influence of two fluoroquinolone derivatives, ofloxacin and pefloxacin, on lymphocyte immunoglobulin secretion and on polymorphonuclear leucocyte superoxide anion production was assessed. Pharmacological concentrations did not influence the studied functions, while suprapharmacological concentrations decreased immunoglobulin production.

Lymphocyte Subclasses and Immunoglobulins in Adults Receiving Radiation Treatment in Infancy for Thymic Enlargement

One-hundred and fifty-three subjects who were irradiated in infancy for an alleged enlarged thymus and 51 controls were studied to quantify the phenotypically different subpopulations of peripheral blood lymphocytes and immunoglobulins. These subjects were selected from a larger cohort which has been followed prospectively since the early 1950s. Previous studies of this cohort have shown a radiation dose-related increased risk of thyroid carcinomas and adenomas in the irradiated group as well as a recent excess of various extrathyroid neoplasms. In this substudy, no significant differences in lymphocyte subpopulations or immunoglobulin levels were observed between the irradiated and control groups. These findings support a hypothesis that the radiation-related excess of neoplasms in this cohort may have resulted from direct cellular damage with subsequent mutations rather than impaired immune function.

Lymphocyte immunoglobulin secretion during short-term treatment with cephalosporins in vivo and in vitro: effects on plaque-forming cell response of human peripheral blood lymphocytes.

The influence of 4 cephalosporins on the plaque-forming cell (PFC) generation in mitogen-activated cultures of human lymphocytes (PBL) was assessed. In vitro, suprapharmacological concentrations of the drug resulted in a decreased PFC response, while pharmacological concentrations did not influence the generation of PFC. Cells exposed to drugs for 24 h in vivo and propagated in autologous serum in vitro showed a decreased PFC response.

Applications and limitations of peripheral blood lymphocyte immunoglobulin light chain analysis in the evaluation of non-Hodgkin's lymphoma.

Surface immunoglobulin (sIg) light chain analysis of peripheral blood lymphocytes (PBL) from 63 patients with non-Hodgkin's lymphoma (NHL) was performed to determine the ratio of kappa-bearing lymphocytes to lambda-bearing lymphocytes (kappa/lambda ratio). In 43% an abnormal kappa/lambda ratio was detected, implying the presence of a malignant clone in the peripheral blood (clonal excess). In 67% of cases with an abnormal kappa/lambda ratio, the absolute lymphocyte count was within normal limits and 20% did not have morphologic bone marrow involvement. Light chain analysis of bone marrow aspirates was performed in three of four patients with a circulating clone and normal bone marrow morphology. All three patients had abnormal bone marrow aspirate kappa/lambda ratios. The presence of a circulating clone was associated with morphologic bone marrow involvement (P less than 0.05). Nine patients with documented B-cell NHL were followed with repeated examinations. The presence of a circulating clone persisted in two patients refractory to therapy and in two patients otherwise believed to be in remission. Conversion to an abnormal kappa/lambda ratio occurred in two patients coincident with the development of new bone marrow involvement and in a third patient prior to the onset of frank leukemia. In three instances, the PBL light chain analyses remained within normal limits in patients with stable lymphadenopathy. The kappa/lambda ratio returned to normal in one patient responding to treatment with a partial remission, and remained within normal limits despite progression of lymphadenopathy in one patient, and local disease recurrence in another patient. These findings suggest that the detection of a circulating clone by sIg light chain analysis may be useful as a noninvasive approach to identify disseminated disease activity unsuspected on the basis of morphologic evaluations, but may not reflect the presence, progression, or recurrence of localized tissue lesions.

Inhibition of human B lymphocyte differentiation by a stable metabolite of cyclophosphamide (ASTA Z 7557, INN mafosfamide).

We have evaluated the immunosuppressive effect of a new stable derivative of cyclophosphamide (ASTA Z 7557) on human lymphocyte immunoglobulin biosynthesis in vitro. When graded amounts of the drug are added to lymphocyte cultures stimulated with the helper T cell-dependent activator (PWM), a marked inhibition of B cell proliferation and differentiation occurs. Lymph node cells are particularly sensitive to the drug, while splenocytes are relatively resistant. The agent exerts only a minor effect on immunoglobulin synthesis triggered by a direct B cell stimulant (S. paratyphi bacteria).

Lymphocyte immunoglobulin receptors in newborn piglets

Lymphocyte immunoglobulin receptors in newborn piglets

Rosette tests for T and B lymphocyte immunoglobulin

B lymphocytes synthesize and then express in their surface membranes immunoglobulin (Ig) molecules which serve as antigen receptors. Although controversy surrounds the precise nature of antigen receptors in T lymphocytes, these receptor molecules react with certain anti-Ig reagents and are considered to be Ig-related. Antiglobulin rosette tests and in particular the mixed antiglobulin rosetting reaction (MARR) and the direct antiglobulin rosetting reaction (DARR) have special merit in demonstration of lymphocyte Ig and Ig-related surface determinants; these lest systems are reviewed and compared with more conventional direct binding methodology which uses free anti-Ig molecules to sensitize lymphocytess.

Effect of adult thymectomy on the development of 1-chloro-2,4-dinitrobenzene contact sensitivity and other T lymphocyte functions in patients with myasthenia gravis.

1-Chloro-2,4-dinitrobenzene (DNCB) contact sensitivity and other T lymphocyte functions were studied in thymectomized and non-thymectomized patients with myasthenia gravis. The ability to develop contact sensitivity was reduced in patients with myasthenia gravis and was further reduced after thymectomy. Memory lymphocyte function, as measured by skin tests with common microbial antigens, was intact. A positive phytohemagglutinin (PHA) skin test was found mainly in those patients who also developed contact sensitivity to DNCB. The number of rosette-forming T cells in the peripheral blood as well as mitogen stimulation with PHA was found to be normal in thymectomized as well as non-thymectomized patients. In the thymectomized group, mitogen stimulation with concanavalin A and staphylococcal protein A was also within the normal range, while increased stimulation was obtained with pokeweed mitogen (PWM) and purified protein derivative (PPD). No alteration in mixed lymphocyte culture reactivity or immunoglobulin levels was obtained compared with healthy blood donors. On the basis of these results, it is concluded that non-thymectomized as well as thymectomized patients with myasthenia gravis may have a defective subpopulation of T cells possibly residing in the TH1-positive population. Furthermore, the increased lymphocyte stimulation obtained with PPD and PWM may indicate a reduction of suppressor cell activity after thymectomy.

Colchicine inhibits phosphatidylinositol turnover induced in lymphocytes by concavalin A.

RECENTLY colchicine has been shown to affect various membrane properties such as the lateral mobility of receptors and other surface membrane markers and the agglutination of different cell types. Many of the described phenomena also involve concanavalin A (con A); for example, phagocytosis by rabbit polymorphonuclear leukocytes (PMNs) leads to a selective depletion of con A binding sites which is prevented by colchicine1, con A inhibition of lymphocyte immunoglobulin receptor capping is reversed by colchicine2–4 and con A-induced aggregation of PMNs, fibroblasts and hepatoma cells is inhibited by colchicine5–7. In addition to these effects on membrane properties, colchicine also has been shown to inhibit con A-induced lymphocyte transformation at an early stage in the sequence of events leading to thymidine incorporation and blast transformation8,9. These various considerations led us to determine if colchicine could alter phosphatidylinositol (PI) turnover in lymphocytes since it is known that increased PI turnover is an early event induced by con A and other mitogenic lectins10–12.

Extrahepatic Malignancy in Chronic Liver Disease: Report of Six Cases

Six patients are described with chronic liver disease, 3 with primary biliary cirrhosis and 3 with chronic active hepatitis, all of whom developed malignant disease in organs other than the liver. Two malignancies were mixed histiocytic-lymphocytic lymphomas. All patients had received corticosteroid or azathioprine therapy. Specific lymphocyte counts and immunoglobulin and antibody determinations failed to reveal a consistent pattern of abnormality to recognize the patient at risk. Several diseases in which the immune responsiveness is disturbed seem to have this potentiality in common, especially if therapy that further alters immunological reactivity is given.

Comparison of immunoglobulin formation in vitro by leukocytes of normal donors and steroid- and nonsteroid-treated asthmatic patients

To characterize further our observed differences between asthmatic patients and normal adults with regard to lymphocyte immunoglobulin (Ig) formation in vitro, we studied the response of lymphocytes from steroid-treated asthmatic patients to hydrocortisone, isoproterenol, and aminophylline. Cells from these donors showed no response to either hydrocortisone or aminophylline, but significantly enhanced Ig formation with isoproterenol (10 −9 to 10 −6 M). Experiments with lymphocytes from nonsteroid-treated asthmatic patients showed no response to isoproterenol, but significantly enhanced Ig formation with aminophylline (10 −7 to 10 −4 M). Corresponding experiments with cells from normal adults showed significantly enhanced Ig formation with both isoproterenol (10 −9 to 10 −6 M) and aminophylline (10 −7 to 10 −5 M). To explore the possible mediation of Ig formation by cyclic 3′,5′-adenosine monophosphate (cAMP), we studied: (1) combinations of aminophylline and isoproterenol, (2) cAMP, and (3) dibutyryl cAMP. Cells from nonsteroid-treated asthmatic patients showed significantly enhanced Ig formation with combinations of aminophylline and isoproterenol, and this enhanced Ig formation was greater than that observed with aminophylline alone. In similar experiments with cells from normal subjects, the enhanced Ig formation with the combinations was of the same magnitude as that with aminophylline alone. Cells from nonsteroid-treated asthmatic patients showed no response to cAMP. Cells from normal subjects had significantly enhanced Ig formation with two concentrations of cAMP (10 −6 and 10 −5 M) and no response to dibutyryl cAMP. Although our data do not add substantial support to the possible mediation of Ig formation by cAMP, they do demonstrate striking differences in response of lymphocytes from asthmatic patients and normal subjects.

QUANTITATICN OF LYMPHOCYTE IMMUNOGLOBULIN BIOSYNTHESIS

Quantitaticn of peripheral blood lymphocyte immunoglobulin biosynthesis was evaluated as a method for detection of humoral immunodeficiency. Lymphocytes isolated from 3-15 cc of heparinized blood, using Ficoll-Hypaque gradients, were incubated in leucine free medium (1×106 cells/cc) with tritiated leucine for 48 and 72 hrs. Newly synthesized secreted IgG and total immunoalobulins were measured using specific anti-immunoglobulin antibodies coupled to insoluble bromacetylcellulose. Synthesis was readily detected in newborn infants as young as 28 wks gestation. The normal range (6 individuals) for total immunoglobulin synthesis and IgG synthesis was 1.3-5.5%, and 0.3-1.6%, respectively, of total secreted proteins. The lymphocytes of a patient with X-linked agammaglobulinemia synthesized no immunoglobulins. In a patient with non-X-linked agammaglobulinemia (with B-cells), total immunoglobulin and IgG synthesis was 0.20% and 0.13%, respectively. A patient with common variable hypogammaglobulinemia (normal IgM, low IgG, absent IgA) also showed depressed synthesis (total immunoglobulins 0.70%, IgG 0.40%). Quantitation of lymphocyte immunoglobulin biosynthesis appears to be a useful technique for the early diagnosis of humoral immunodeficiency and may be of potential value for the detection of immunodeficiency disease carriers.

Restriction of the mobility of lymphocyte immunoglobulin receptors by concanavalin A.

The induction by anti-immunoglobulin of patch and cap formation on mouse lymphocytes was inhibited by the addition of concanavalin A. This effect was reversed by alpha-methyl-D-mannoside, a competitive inhibitor of the binding of concanavalin A. Concanavalin A prevented both patch and cap formation, in contrast to the metabolic inhibitor NaN(3), which prevented only cap formation. This suggests that binding of concanavalin A induces changes on or in the lymphocyte membrane that inhibit free diffusion of immunoglobulin receptors.