Fc Receptor-bearing Lymphocytes Research Articles

Intact nuclear membrane in necrotic keratinocytes of Stevens-Johnson syndrome distinguishes complement-mediated target cell lysis in ADCC from cytotoxic T-lymphocyte-mediated cytolysis

Isolated keratinocyte necrosis, often visible even on light microscopy, is the end result of target cell lysis resulting from several immune mechanisms. On electron microscopy, the keratinocyte necrosis may be distinguished by the concomitant presence or absence of an intact nuclear membrane. This morphological feature allows the deduction of the mechanism responsible for target cell damage. Thus, isolated keratinocyte necrosis with loss of integrity of the nuclear membranes, often associated with calcium deposits within the nucleus, occurs as a result of cytotoxic T-lymphocyte-mediated cytolysis. On the other hand, isolated keratinocyte necrosis with intact nuclear membranes occurs as a result of complement-mediated damage either from antibody-dependent cell-mediated cytotoxicity or from complement-based membrane attack complex per se. However, in the latter, T lymphocyte subsets are absent. In Stevens-Johnson syndrome, the intact nuclear membrane of necrotic keratinocytes suggests that complement- mediated mechanisms are involved, while the identification of lymphocytes bearing Fc receptors (leu 11b +) for immunoglobulin, implicates antibody-dependent cell-mediated mechanisms, i.e. antibody-dependent cytotoxicity, in the pathophysiology of the disease.

Allergen‐directed expression of Fc receptors for IgE (CD23) on human T lymphocytes is modulated by interleukin 4 and interferon‐γ

T lymphocytes bearing Fc receptors (FcR) for immunoglobulins are known to have immunoglobulin class-specific regulatory functions. Here we report that expression on T cells of the low-affinity FcR for IgE (Fc epsilon RII/CD23) is preferentially induced by stimulation with antigens that cause an IgE response. T cells from eight patients allergic to the hemoglobin of Chironomus thummi thummi mosquito larvae (CHIT I) were analyzed for reactivity with the anti-FcERII/CD23 monoclonal antibody (mAb) M-L25 under various conditions. No Fc epsilon RII/CD23+ T cells were observed among freshly isolated, resting peripheral blood mononuclear cells (PBMC). Stimulation of PBMC with CHIT I, however, induced a marked although transient Fc epsilon RII/CD23 expression on a large portion of the allergen-activated T lymphocytes. It reached a maximum of 37.2 +/- 4.6% Fc epsilon RII/CD23+ T cell blasts on day 5 of culture. The selectivity of this expression became evident when compared to non-allergenic control antigens: after stimulation of PBMC with tetanus toxoid or purified protein derivative from tuberculin a maximum of 4.6% +/- 1.4% and 4.2% +/- 1.1% T cell blasts was found to express Fc epsilon RII/CD23, respectively. Activation by an anti-CD3 mAb was insufficient to induce Fc epsilon RII/CD23 on T cells. The allergen-stimulated Fc epsilon RII/CD23+ T cells exclusively belonged to the CD4+CD29+ helper inducer T cell subset. Using a cDNA probe coding for the B cell Fc epsilon RII/CD23, Northern blot analysis revealed a 1.7-kb Fc epsilon RII/CD23 mRNA in extracts of highly purified allergen-stimulated T cells. It was of the same size as Fc epsilon RII/CD23 mRNA of the lymphoblastoid B cell line WI-L2. Of several cytokines tested [interleukin (IL) 1 to IL 6, interferon-gamma (IFN-gamma), tumor necrosis factor-alpha] only IL 4 and IFN-gamma significantly modified allergen-induced Fc epsilon RII/CD23 expression on T cells. The latter was enhanced nearly twofold in the presence of IL 4, and was almost completely abrogated by IFN-gamma. IL 4, however, could not increase the number of Fc epsilon RII/CD23+ T lymphocytes either alone or in combination with an anti-CD3 mAb. Taken together, the selective induction of Fc epsilon RII/CD23 on T cells by allergen and its inclusion in the regulatory network of cytokines point to an important role of Fc epsilon RII/CD23+ T lymphocytes in the human IgE response.

Protein malnutrition alters the distribution of Fc gamma R+ (T gamma) and Fc mu R+ (T mu) T lymphocytes in experimental pulmonary tuberculosis

Inbred, strain 2 guinea pigs were given isocaloric diets containing either 30% (control diet) or 10% (low-protein diet) ovalbumin and infected 4 weeks later by the respiratory route with virulent Mycobacterium tuberculosis. By using an Fc receptor rosette assay, the proportions of T lymphocytes bearing Fc receptors for immunoglobulin G (T gamma cells) or immunoglobulin M (T mu cells) were quantified in blood and lymphoid tissues taken postinfection. A significant elevation in the proportion of the putative suppressor T subset (T gamma) in the blood of protein-deprived guinea pigs was observed at all intervals postinfection. Conversely, the levels of the putative helper T subset (T mu) in the bronchotracheal lymph nodes draining the site of virulent infection in malnourished animals were significantly reduced. Diet did not influence T gamma or T mu cells in the spleens. Diet-induced loss of purified protein derivative-specific T-cell functions in tuberculosis may be associated with alterations in the proportions of or the balances between T gamma and T mu subsets.

Vaccination with Mycobacterium bovis BCG affects the distribution of Fc receptor-bearing T lymphocytes in experimental pulmonary tuberculosis

Inbred strain 2 guinea pigs were vaccinated with Mycobacterium bovis BCG or were left unvaccinated and challenged 6 weeks later by the respiratory route with virulent Mycobacterium tuberculosis. By using a double rosette assay with isotype-specific antibody-coated ox and uncoated rabbit erythrocytes, the proportions of T lymphocytes bearing Fc receptors for immunoglobulin G (IgG) (T gamma cells) or IgM (T mu cells) were quantified in tissues taken from animals that were killed within 4 weeks postchallenge. Tuberculin reactivity in vivo and in vitro and antimycobacterial resistance were also measured. BCG vaccination protected the guinea pigs and resulted in significantly enhanced proportions of T mu cells in the blood during the first 3 weeks and in the spleen during weeks 2 and 3 postchallenge. Levels of T gamma cells declined in all tissues during the first 3 weeks of infection and were unaffected by prior vaccination with BCG. Increased proportions of T mu cells in the blood were accompanied by dramatic tuberculin skin reactions and purified protein derivative-induced lymphoproliferation in BCG-vaccinated guinea pigs during the first 2 weeks following virulent pulmonary challenge. Peak levels of T mu cells in the spleens of vaccinated animals at 2 weeks coincided with the first appearance of virulent mycobacteria in that organ. BCG vaccination appears to influence immunoregulatory events in pulmonary tuberculosis through effects on the distribution of IgM Fc receptor-bearing (T mu cell) T lymphocytes.

Increased natural killer cell activity in sarcoidosis.

We have studied NK cell activity in 32 patients with sarcoidosis and in 29 control subjects. Cytotoxicity was increased in both active and inactive sarcoids, the lytic activity of sarcoid peripheral blood mononuclear cells (PBMC) being approximately 50% higher than that of control subjects. The number of lymphocytes bearing Fc receptors for IgG (FcR-IgG) was correlated with NK cell activity, both parameters being increased in the sarcoid group. Sarcoid NK cell activity was reduced to control levels after depletion of plastic adherent cells, whilst inhibition of cytotoxicity by PGE2 was increased using non-adherent sarcoid PBMC. Plastic depletion had no effect on NK cell activity or its inhibition by PGE2 using PBMC from control subjects. The results demonstrate the presence of a weakly adherent, PGE2 insensitive NK cell in sarcoid PBMC preparations.

Increased T gamma and T mu cells in BALB/c mice with IgG and IgM plasmacytomas and hybridomas.

The results of previous studies in our laboratory have shown that mice bearing plasmacytomas and hybridomas that secrete IgA or IgE are accompanied by increased frequencies of Lyt-1-2+ T lymphocytes bearing Fc receptors (FcR) for IgA (T alpha) or IgE (T epsilon), respectively. The present study was undertaken to examine whether IgG- or IgM-secreting tumors influenced the frequency of T lymphocytes that express FcR for IgG or IgM. We studied mice bearing IgG- and IgM-secreting plasmacytomas and hybridomas. BALB/c mice injected subcutaneously with the IgG-secreting hybridoma HDP1 (gamma 1 kappa, anti-TNP) were sequentially examined for the frequencies and Lyt phenotypes of splenic lymphocytes bearing FcR for IgG (T gamma), IgM (T mu), and IgA (T alpha). A threefold increase in the frequency of T gamma lymphocytes that were Lyt-1-2+, L3T4- was seen. The frequencies of T mu and T alpha lymphocytes in these mice were not significantly altered. Similarly, mice injected subcutaneously with the IgM-secreting plasmacytoma MOPC 104E (mu lambda, anti-dextran) or the IgM-secreting hybridoma C1D1 (mu kappa, anti-ox RBC) were examined sequentially for the frequencies of T gamma, T mu, and T alpha lymphocytes. Mice with established IgM subcutaneous tumors showed a twofold increase in splenic, nylon wool-nonadherent T mu lymphocytes. This was associated with a relative increase in Lyt-2+ splenic T lymphocytes and a relative decrease in Lyt-1+ splenic T lymphocytes. No changes were observed in the frequencies of either T gamma or T alpha lymphocytes. These studies extend to IgG and IgM the observation that plasmacytomas and hybridomas secreting immunoglobulins of a specific isotype cause an expansion of T lymphocytes bearing FcR specific for the corresponding isotype. The expansion of FcR+ Lyt-1-2+ T lymphocytes likely represents an exaggerated, but otherwise normal, immunoregulatory response of the host. These cells may be an important element in the regulation of isotype expression.

Effect of plasmapheresis on E-rosette receptors in SLE patients.

The present study reports on the effects of plasmapheresis on both cellular (E-rosette receptor- and Fc receptor-bearing lymphocytes) and humoral (antilymphocyte antibodies) immunity in patients with active SLE. Ten untreated patients were studied before and after two plasma exchanges. The E-rosette receptor-positive cells were studied by OKT11 monoclonal antibody and Fc receptor-bearing cells by Leu15 monoclonal antibody. Antilymphocyte antibodies were studied before and after plasma exchange by an indirect immunofluorescence technique on autologous and normal lymphocytes. A reduction in OKT11+ and Leu15+ lymphocytes was observed in all patients before plasmapheresis. Antilymphocyte antibodies were detected in five patients. After plasma exchange, an increase in E-rosette receptor- and Fc receptor-bearing lymphocytes was observed along with a reduction in the amount of antilymphocyte antibodies.

Cytochemical distribution of dipeptidylaminopeptidase IV (DAP IV; EC 3.4.14.5) in T-lymphoblastic lymphoma/leukemia characterized with monoclonal antibodies

In human blood and bone marrow, dipeptidylaminopeptidase IV (DAP IV; EC 3.4.14.5) selectively occurs in T lymphocytes bearing Fc receptors for IgM. In the present study 35 cases of lymphoblastic lymphoma and leukemia were analysed for the specificity, incidence and reaction pattern of DAP IV. On the basis of immunohistochemical staining with monoclonal antibodies and enzyme cytochemical staining for acid phosphatase, 12 cases were classified as B-type neoplasms. In 23 cases T-cell properties were expressed to different extents, apparently reflecting different categories of maturation. Whereas B-cell lymphomas were invariably negative for DAP IV, seven of the 23 T-lymphoblastic lymphomas/leukemias showed this enzyme. Thus DAP IV is a highly specific marker for a distinct T-cell subpopulation, apparently irrespective of the stage of differentiation.

Characterization with monoclonal antibodies of T lymphocytes bearing Fc receptors for IgE (T epsilon cells) and IgG (T gamma cells) in atopic patients.

Peripheral blood T lymphocytes from nonatopic control donors, asymptomatic atopic donors, and patients with moderate to severe atopic dermatitis were analyzed for Fc receptors for IgE (T epsilon cells) and IgG (T gamma cells) by rosette assays and were characterized with monoclonal antibodies. The T cells were reacted first with monoclonal antibodies, followed by fluoresceinated F(ab')2 goat antimouse Ig; they were then rosetted, and subsequently the rosetting cells were examined for immunofluorescence. Seven nonatopic control donors had less than 0.1% T epsilon cells and a mean +/- SD of 10.5% +/- 4.1 T gamma cells. Seven asymptomatic atopic donors with low IgE levels (2 to 233 IU/ml) varied from less than 0.1 to 1.3% T epsilon cells and 7.2% +/- 3.7 T gamma cells. Six of seven patients with moderate to severe atopic dermatitis and IgE levels of 1339 to 24,261 IU/ml had less than 0.1% T epsilon cells and significantly fewer T gamma cells (3.1% +/- 2.7, p less than 0.01) than the nonatopic control donors and the atopic donors in remission. Both T epsilon and T gamma cells reacted with the pan-T cell antibody Lyt-3 (anti-sheep red cell receptor) but not with antibodies OKT3, OKT4, or OKT6. Subpopulations of both T epsilon and T gamma cells reacted with antibodies OKT8 and the antimonocyte antibody OKM1. The OKM1+ cells did not appear to be monocytes, however, because the T cells did not react with another antimonocyte antibody, BRL.2, and were negative for nonspecific esterase activity. (ABSTRACT TRUNCATED AT 250 WORDS)

Inhibition of lymphocyte response by prostaglandin-producing suppressor cells in patients with melanoma.

Peripheral blood mononuclear cells (PBMC) from 11 patients with metastatic melanoma (group II) had a significant decrease in blastogenesis to concanavalin A (Con A) (38.7 +/- 7.7 cpm X 10(3); mean +/- SE) compared to 21 patients who were disease free (70.6 +/- 6.7) or 16 healthy controls (83.6 +/- 10.3). If PBMC from patients were preincubated for 72 hr prior to exposure to mitogen, blastogenesis was restored to normal. In group II patients a similar improvement in reactivity of fresh PBMC occurred with indomethacin addition or following rigorous depletion of adherent monocytes. Supernatants from cells cultured with and without Con A in several group II patients contained very high levels of endogenous PGE2. Patients had a greater percentage of T lymphocytes bearing Fc receptors for ox erythrocytes (T gamma) than controls, which appeared to correlate with the increased sensitivity to suppression by exogenously added PGE2. These data suggest that the decreased blastogenesis in certain melanoma patients is due to an increase in PGE2 production by monocytes, along with an increase in lymphocyte sensitivity to its effects.

Measurement of K cell activity: Use of human OR 1R 1 red cells and anti-D to study the role of different leukocytes as effector cells

A method for quantitative assay of K cell activity (KCA) with an allogenic combination of human OR 1R 1 red cells and anti-D (IgG, incomplete antibody) as target cells is described. Lymphocytes alone have been shown to cause lysis of these target cells. Fc receptor-bearing lymphocytes appear to be the chief mediators of KCA. This sub-population is a heterogeneous mixture of sub-sets of other sub-populations defined on the basis of different surface markers and likely to show variable expression of Fc receptors by the EAγ rosette method. Monocytes and neutrophils (and possibly eosinophils), most of which bear Fc receptors, are ineffective as effector cells. Monocytes may in some instances have an inhibitory effect on the expression of KCA of lymphocytes. The method described has the advantage of simplicity (easy availability of OR 1R 1 cells and anti-D) and a high degree of sensitivity (marked lysis at low L/T ratio) and appears suitable for routine use in clinical practice.

Studies on the cellular nature of Candida albicans-induced suppression.

Animals treated with formalinized Candida albicans manifest depressed cellular immune activity. Splenocytes from mice treated with as little as 14 micrograms of this material exhibited significantly reduced responses to the T cell-dependent mitogens phytohemagglutinin and concanavalin A. On the other hand, the B lymphocyte-dependent response to bacterial lipopolysaccharide was normal in these cultures. Splenocytes from treated mice were capable of actively suppressing the T cell- (but not B cell-) dependent proliferative response of normal cells. Analysis of splenocytes from Candida-treated mice showed that the suppressor cell is adherent to glass wool, is not adherent to Sephadex G-10, does not phagocytize carbonyl iron, is not susceptible to treatment with anti-Thy-1 plus C, but does bind specifically to anti-immunoglobulin- (anti-Ig) coated dishes. The adherence to the anti-Ig-coated dishes was not due to the simple attachment of Fc receptor-bearing lymphocytes, because dishes coated with the F(ab')2 fragment of rabbit antimouse IgG bound the suppressor cell. These results suggest that the active Candida-induced suppressor cell is composed, at least in part, of surface Ig-bearing B lymphocytes.

Suppressor T cells are activated in vivo in patients with multiple sclerosis coinciding with remission from acute attack.

By a variety of assays, investigators (1-4) from several laboratories previously observed changes in numbers of thymus-derived suppressor lymphocytes (Ts) in patients with acute active multiple sclerosis (MS). Changes in numbers of Ts in the peripheral blood compartment of such patients closely follow disease activity. Extending these observations we now report Ts are activated in the peripheral blood of certain patients with MS. This activation occurred in vivo in the majority of patients with active disease, but only during remission after an acute episode of MS. In contrast, activated Ts were found infrequently in patients with chronic progressive or stable MS, with neurologic diseases other than MS, or healthy individuals. The suppressor activity found in peripheral blood T cells during the acute remitting stage of disease was associated with elevated numbers of lymphocytes bearing Fc receptors for IgG and/or OKT8 markers.

Medullasin enhances human natural killer cell activity.

Medullasin, a new serine protease found in bone marrow cells, increased markedly human natural killer cell activity. Whereas the natural killer cell activity measured immediately after the treatment with medullasin remained almost on the same level as the control, an incubation at 37 degrees C for several hours increased markedly the natural killer cell activity of the lymphocytes treated with medullasin. Enhancement of the natural cytotoxicity was caused by the treatment with physiologic concentrations of the protease (5-20 micrograms/ml). Inhibitors of medullasin such as phenylmethylsulfonyl fluoride and elastatinal prevented the activation of natural cytotoxicity. Depletion of lymphocytes bearing Fc receptors for IgG abolished the enhancement of natural killer cell activity by medullasin. Interferon activity was not detected in the supernatant of lymphocyte cultures stimulated with medullasin. The medullasin enhanced further the natural killer cell activity of lymphocytes stimulated with interferon. Medullasin activity was detected neither in unstimulated nor stimulated (by concanavalin A or phytohemagglutinin) human lymphocytes. The protease was released easily from human mature granulocytes into culture medium. It is considered from these results that the level of human natural killer cell activity is regulated by medullasin released by mature granulocytes.

Activation of Fc receptor-bearing lymphocytes by immune complexes. I. Stimulation of lymphokine production by nonadherent human peripheral blood lymphocytes.

Activation of human peripheral blood lymphocytes by incubation with particulate immune complexes or aggregated human gamma-globulin was studied by measuring the release of leukocyte migration inhibitory factor (LIF) activity. LIF-active supernatants were consistently produced when nonadherent lymphocytes containing less than 1% surface immunoglobulin-bearing cells and less than 0.2% nonspecific esterase-positive monocytes were incubated in the presence of RBC sensitized with rabbit or human antibodies or with pooled heat-aggregated human gamma-globulin. This immune complex-induced lymphokine production (ICLP) was dependent on the presence of cells bearing receptors for the Fc portion of IgG (Fc gamma). ICLP could not be demonstrated with lymphocyte preparations enriched for B cells even though the latter showed vigorous LIF production in the presence of complement-sensitized erythrocytes. ICLP was dependent on the concentration of lymphocytes and of stimulant as well as on the duration of coincubation, and it required active metabolic processes and RNA and protein synthesis but not DNA synthesis. Ca++ but not Mg++ was obligatory. ICLP by non-B Fc gamma receptor-bearing lymphocytes may play a role in antibody-dependent protective inflammation and immunologic injury phenomena, which is similar to that of lymphokine release by antigen-activated T cells in delayed hypersensitivity responses.

Immunological Monitoring of a Patient with Guillain-Barré Syndrome Successfully Treated with Plasma Exchange

A patient with Guillain-Barré syndrome was successfully treated by plasma exchange (without any other therapy), and the relationship between changes in his peripheral blood lymphocyte subpopulations and the clinical improvement induced by plasmapheresis was studied. At the recovery, the most interesting findings were an increase in T suppressor lymphocytes, and in Fc receptor-bearing lymphocytes (above normal values), together with a decrease in B lymphocytes. In this patient, plasma exchange not only removed toxic humoral factors, but also restored a normal balance among peripheral lymphocytes.

Histochemical patterns of human T lymphocyte subpopulations with nonspecific esterase staining.

Alpha-naphthyl-acetate-esterase (ANAE) activity was histochemically demonstrated in human T lymphocytes and their subpopulations. Examination of T lymphocytes bearing Fc receptors for IgM showed the presence of circumscribed deposits of esterase activity, while T cells with FC receptors for IgG showed of predominantly diffuse granular pattern. The difference in the topography of the enzymatic activity was shown to be related to cellular activation resulting from exposure to IgG-sensitized bovine erythrocytes or to strong polyclonal activation by lectin mitogens. The ANAE stain also characterized monocytes, which exhibited an intense nongranular cytoplasmic stain. Non-T cells showed no detectable ANAE activity when tested by this method.

Activation of Fc receptor-bearing lymphocytes by immune complexes. II. Killer lymphocytes mediate Fc ligand-induced lymphokine production.

Cells that participate in immune complex-induced production of leukocyte migration inhibitory factor (LIF) activity can be concentrated in a population making up 2-4% of human peripheral blood lymphocytes in which greater than 90% of the cells are active in a single cell antibody-dependent cellular cytotoxicity assay. When so concentrated, such killer (K) cell preparations are as efficient in producing LIF activity as mitogen activated T lymphocytes. Other Fc receptor (FcR)-bearing lymphocytes, including natural killer (NK) cells, do not produce measurable LIF activity when incubated with immune complexes (additional evidence that the K and NK cells among ligands to the FcR of the appropriate lymphocytes, possibly without need for exogenous receptor bridging, is the only requirement for their activation to immune complex-induced lymphokine production (ICLP). It is probable that ICLP by K cells palays a role in antibody-mediated effector functions in vivo.

Lymphocytes bearing Fc receptors for IgE. VI. Suppressive effect of glucocorticoids on the expression of Fc epsilon receptors and glycosylation of IgE-binding factors.

Incubation of rat lymphocytes with homologous IgE induced an increase in Fc epsilon R(+) lymphocytes and the formation of IgE-binding factors. Pretreatment of rat lymphocytes with 1 to 5 microM dexamethasone, however, prevented the IgE-induced expression of Fc epsilon R on both B and T lymphocytes. Upon incubation with IgE, T cells activated with 10 micrograms/ml Con A produced IgE-potentiating factors that had affinity for lentil lectin and selectively enhanced the IgE response. Pretreatment of the Con A-activated T cells with dexamethasone, before incubation with IgE, changed the nature of IgE-binding factors formed by the cells. The majority of IgE-binding factors formed by the dexamethasone-treated, Con A-activated cells failed to bind lentil lectin Sepharose and selectively suppressed the IgE response. An injection of 0.2 mg dexamethasone into rats infected with Nippostrongylus brasiliensis markedly diminished the proportion of Fc epsilon R(+) lymphocytes in their mesenteric lymph nodes. The lymph node cells from the infected animals spontaneously released IgE-potentiating factors in vitro. However, the majority of IgE-binding factors formed by the mesenteric lymph node cells from the dexamethasone-treated, Nb-infected animals lacked affinity for lentil lectin and selectively suppressed the IgE response. The results indicate that glucocorticoid treatment prevented the glycosylation of IgE-binding factors and thereby changed the biologic activities of the factors.

Lymphocytes bearing Fc receptors for IgE. VII. Possible participation of phospholipase A2 in the glycosylation of IgE-binding factors.

Lymphocytes bearing Fc receptors for IgE. VII. Possible participation of phospholipase A2 in the glycosylation of IgE-binding factors.